Download p53 in Hematologic Malignancies

From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
REVIEWARTICLE
p53 in Hematologic Malignancies
By Jun Imamura, lsao Miyoshi, and H. Phillip Koeffler
T
HE p53 HAS BEEN CHOSEN as molecule of the year
for 1993 by the journal Science.' The protein's illustrious history began quietly in 1979, when a 53-kD eukaryotic
protein was
shown
to
bind
to
SV40 large
T
The
protein was named p53 becauseofitssize.Initial
studies
suggested that p53 was an oncogene because it could transform rodent ~ e l l s . ~The
- ' ~p53 genes used in these studies
hadbeenisolated from cancer cell lines;these p53 genes
were subsequently discovered to containmissense mutations
andthe resultant mutant proteins had propertiesdifferent
from thoseofwild-type
(wt) ~ 5 3 . ' . ' ~Investigators
"~
also
noted that several virally transformed
murine cell lines as
well as a human myeloid leukemic cell line (HL-60) had
major deletions of the p53 gene.",'" We found that the p53
gene was mutated in human cancer; four of five osteosarcomas frompatients had major disruptionsof their p53 genes."
Other studies foundthat loss of heterozygosity (LOH) of the
short arm of human chromosome 17 in the region coding for
p53 occurred in a number of human cancers.22-24 Additional
careful analysis of the p53 gene has
shownthat it is frequently mutated in more than 50 varieties of human cancers
including lung, breast, thyroid, gastrointestinal, and ovarian
cancers, lymphomas/leukemias,and brain tumors.2',2s-44
Furtherfunctional studies determinedthat wt-p53 suppressed
transformation of cells's"sand overexpression of wt-p53
blocked cells in the G , phase of the cell cycle.J',46 Taken
together, p53 fulfills the criteria of a tumor-suppressor gene,
including the finding of LOH in the region of p53 in tumors,
the presence of p53 mutations in human and murine tumors
and transformed cell lines, and the ability of wt-p53 to suppress transformation of cells having p53 mutations. In one
decade, a protein discovered through an effort to understand
how SV40 transforms cells was initially characterized as an
oncogene, only to find that it is a pivotal tumor-suppressor
that is the guardian of DNA-damaged cells by halting their
proliferation, pushing badly damaged cells into an apoptotic
cell death and preventing unwanted DNA amplification.
This reviewwill characterize the p53 abnormalities in
hematopoietic malignancies and discuss the clinical significance of these alterations. In addition, potential therapeutic
approaches will be briefly mentioned.
DETECTING p53 ABNORMALITIES IN HUMAN CANCERS
Southern blottinganalysis was the first methodused to
detect p53 mutations in various cancers." However, this is
a cumbersome technique that can onlydetect gross alterations. An indirect detection method is the analysis of LOH,
taking advantage of DNA p~lymorphisms.'~ The paradigm
for LOH is that one p53 allele develops a point mutation;
then, through one of several genetic mechanisms, including
recombination or duplication, the normal p53 allele is lost.
Therefore, LOH in the region of a tumor-suppressor gene is
analogous toa tombstone marking thelethally injured tumorsuppressor gene. Several very informative polymorphic sites
are present in the region of the p53 gene.40,4x This indirect
approach of identifying tumors with potential p53 mutations
is flawed because LOH in the region of p53 gene can occur
in the absence of a detectable p53 mutation, perhaps because
another tumor-suppressor gene exists in the same chromosomal region." Also,thisanalysisrequires
normal tissue
from the same individual.
Analysis of single-strandconformationalpolymorphism
(SSCP) using polymerase chainreaction (PCR) allowsa
relativelysimpledetection system for point mutations.4y~54
The PCR-SSCP technique can detect an abnormality in less
than 10% DNA containing mutant p53 in a background of
90% DNA containing ~ t - p . 5 3 . ~ ~The
~ ' ' specificity of PCRSSCP ismore than 95% for 100- to 300-bp PCR fragments.5"
Thus, this protocol is very useful to screen for mutations in
a short regionof a gene. Those sampleswith abnormal SSCP
require nucleotide sequencing toassert if the abnormal SSCP
representsamutationthat
eitheralters an amino acid or
merely represents a polymorphism or silent mutation. A potential false-negative result can occur if the p53 mutation is
located outside the area examined by SSCP. More than 90%
of mutations of p53 probably occur in exons 5 through 8
andmostanalysishas
focused onthis r e g i ~ n . ' ~ . ' " ~An
~-~'
enormous problem is that the SSCP and sequencingtechniques are very time-consuming and require refined expertise. Less frequently used techniques, such as either denaturing gradient gel electrophoresis, RNase protection assay, and
detection of basepair mismatches with hydroxylamine and
osmium tetroxide, have their own inherit problems and usually are even more labor-intensive than PCR-SSCP.sx-62
Immunohistochemistry for p53 is the simplest analysis for
p53 integrity, but this approach cannot directly detect p53
mutations. Because ofashorthalf-life
of about 6 to 20
From the Cedars-Sinai Medical Center/UCLASchool of Medicine,
minutes, wt-p53 usually does not accumulate in most normal
Division of Hematology/Oncology, Los Angeles, CA; and the Dein
amounts detectable by immunohistochemical
partment of Internal Medicine, Kochi Medical School, Kochi, Japan. tissues
Submitted March 16, 1994; accepted June 17, 1994.
methods. However, most missense mutations of p53 prolong
Supported by National Institutes of Health Grants No. CA42710,
the half-life of the protein, permitting it to be immunohistoCA33936, and DK42792 as well as bythe Parker Hughes Leukemia
chemically detectable in those tumors containing a p53 misFund and Concern Foundation.
sense m ~ t a t i o n . ~ ~The
. ~ ' technique
~~"
is rapid and easily perAddress reprint requests to H. Phillip Koefler, MD, Chiej Hemaformed by many
pathology
laboratories.
Nevertheless,
tology/Oncology
Division,
Cedars-Sinai
Medical
CenterIUCLA
quantitation is difficult and false-positives and -negatives
School of Medicine, 8700 BeverlyBlvd, 8210, Los Angeles,CA
can occur dependingon the tissue. False-negatives are partic90048.
ularly the case if the p53 mutation results in either a prema0 1994 by The American Socier): of Hematology.
ture stopcodon, frame-shift, or alteration of a splice site
0006-4971/94/8408-03$3.00/0
2412
Blood, Vol 84, No 8 (October 15), 1994: pp 2412-2421
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
2413
p53 IN HEMATOLOGICMALIGNANCIES
Table 1. p53 Alterations in Hematopoietic Malignancies
Disease
Frequency of Mutation (%)
MDS
AML
C-ALL
ALL L3
T-ALL
T-ALL relapsed
CLL
Richter's syndrome
B-low-grade lymphoma
B-high-grade lymphoma
Burkitt's lymphoma
HD
ATL
CML
Chronic phase
Blast crisis
CTCL
HCL
T-CLL
Multiple myeloma
5
15
3
50
Rare
30
15
40
Rare
30
40
70*
40
Rare
20-30
Rare
10
Rare
5
~
~~
Data are from primary malignancies and do not include data from
cell lines.
Abbreviations: MDS, myelodysplastic syndrome; AML, acute myelogenous leukemia; ALL, acute lymphocytic leukemia; C-ALL, common ALL; ALL LS,Burkitt's type ALL; CLL, chronic lymphocytic leukemia; ATL, adult T-cell leukemia; CML, chronic myelogenous leukemia;
CTCL, cutaneous T-cell leukemia; HCL, hairy cell leukemia.
* Sixty percent to 80% of cases of mixed cellularity and nodular
sclerosing type HD have RS cells that are p53 positive by immunohistochemistry.
or enhancer/promoter region. These types of p53 mutations
represent about 10% of the total p53 alterations. False-positives also can occur. For example, in one series of lymphomas, we found 50% of tumors with immunohistochemically
detectable p53, but no mutations were detected in the p53
gene.7' This finding could be explained in part because we
did not analyze for alterations in every region of the gene.
Also, some rapidly dividing normal tissue express p53, such
as activated T lymphocytes. In addition, p53 might be detected if wt-p53 was bound to another protein that inactivated
but prolonged its half-life (ie, SV40 lzrge T antigen). Ideally,
a p53 antibody that can detect only mutant p53 is required
for immunohistochemistry, but this reagent is not yet available. The monoclonal antibody known as pAb 240 has specificity for many mutant p53s by immunoprecipitation of nondenatured p53, but the antibody cannot differentiate mutant
from wt-p53 on either Western blot or immunohistochemistry because both denature the p53 pr~tein.~'
p53 ALTERATIONS IN HEMATOLOGIC MALIGNANCY
A summary of the incidence of p53 mutations in hematologic malignancies is provided in Table 1. Several of the
notable features of p53 alterations in these diseases are summarized in Table 2 and these features are discussed within
the context of the individual disease.
Chronicmyelogenousleukemia
(CML). The structure
and expression of the p53 gene is altered in about 20% to
30%of samples from patients in myeloid blast crisis of
CML, whereas chronic-phase CML cells only rarely have
detectable p53 a l t e r a t i ~ n s . ~ ~ ~Several
' , ~ * ~ features
' * ~ ~ of p53
and CML are the following: (1) CML is analogous to osteosarcoma in so far as the p53 gene can be altered by either
point mutations or major DNA rearrangements. Why these
two diseases frequently have major rearrangements of p53
is unclear. Another malignancy that usually has a major p53
rearrangement is murine erythroleukemia, which is associated with a viral infection. (2) The p53 alterations almost
always occur in myeloid, not lymphoid, blast crisis. (3) The
p53 mutations are associated most frequently with samples
in which one of the short arms of chromosome 17 (1 7p) has
been loss, usually through formation of either an isochromosome 17q [i(17q)] or unbalanced tran~location.~~
The i(17q)
chromosome occurs in about 30% of cases of myeloid blast
crisis of CML and about 40% of these have p53 mutations
on the remaining p53 allele.74The loss of a 17p (containing
p53) may precede the p53 mutation of the remaining allele
in CML.74 In contrast, the p53 mutations in gliomas and
colorectal and breast tumors occur on one p53 allele and
then the remaining normal p53 allele is lost. In either case,
these observations emphasize the strong selection for complete loss of p53 function in the process of carcinogenesis.
(4) Circumstantial evidence strongly suggests that a p53 mutation in the CML clone can result in disease transformation
to
myeloid
blast crisis?' Pari passu, when wt-p53 is
transfected and stably expressed in the p53 null CML erythroblastic cell line K562, growth of the cells slows and they
undergo partial differentiation suggesting an involvement of
wt-p53 in the differentiation processes.75Although rare, the
finding of a p53 mutation in myeloidcells during the chronic
phase probably is a grave prognostic sign.
Acute myelogenous leukemia (AML) and meylodysplastic
syndrome (MDS). In 1986, we found that 8 of 33 patient
samples from a variety of hematopoietic malignancies
showed increased accumulation of p53 using immunoprecipitation analy~is.'~
Seven of the eight samples occurred in
cells of patients with either preleukemia or AML. These and
similar results suggested the notion that p53 may contribute
Table 2. Notable Features of p53 Alterations
in Hematopoietic Malignancies
1. Development of p53 mutations is often correlated with
worsening or relapsing of the hematopoietic malignancy.
2. Loss of short arm of chromosome 17 is associated with a p53
mutation on the remaining allele in several hematopoietic
malignancies.
3. B-cell lymphomas with p53 mutations often have c-myc
activation, but EBV infection does not appear to correlate with
the presence of a p53 mutation.
4. HD has p53-positive RS cells but accompanied lymphocytes,
eosinophils, and macrophages do not overexpress p53 consist
with these cells being a reaction to the malignant process.
Lymphocyte-predominant HD does not have p53-positive RS
cells.
5. Individuals with LFS have a p53 mutation in their germline and
have an increase incidence of leukemias and lymphomas.
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
2414
to the phenotype of certain leukemias Frequent
in
p53 abnormalities have been reported
in cell lines derived
from acute myeloid leukemia samples, suggesting that p53
gene inactivationmayhavea
role in theestablishmentof
these This
cell
was
first observed in HL-60
cells thathaveamajorrearrangement
of p53 resulting in
absence of p53 expression." Although frequent in AML cell
lines, the mutational frequency
of p53 in AML cells from
individualsabout
is
The mutational
frequency
increases to about 50% in AML samples having 17p monosomy, similar to what is observed in myeloid blast crisis of
IMAMURA, MIYOSHI, AND KOEFFLER
and overexpression of the protein. These tumors appear to
have normal p53 alleles, supportingtherole of mdm-2 in
the inactivation of wt-p53 resulting in tumor progression."
Conceivably overexpressed mdm-2 or another as yet unidentified protein is binding and inactivating p53, which contributes to the process of leukemogenesis. However, evidence
for this hypothesis is presently lacking.
Of note, the introduction of wt-p53 into a murine p53 null
AML line (MI) had no effect on differentiation of the cells,
but induced their apoptosis; this was inhibitable by a growth
factor (interleukin-6 [IL-6])."x Furthermore, myeloid progenCML,X(J.XI
itor cells and thymocytes from pS3-deficient mice are more
The p53 gene is also infrequently altered in MDS, with a
resistant than their normal counterparts to development of
frequency of about 5% to 10%; most of these are missense
apoptosis."".""' Together, theseresultssuggestthatnormal
point mutations.JZ~X2~8"
Characteristics of p53 in theMDS
myeloid progenitor cells might continuously undergo apoclone include the following. (1) p53 mutations occur in the
ptotic death in the absence of appropriate differentiation and
proliferation signals. Loss of p53 in these cells could provide
subtypes of MDS with a prognostically poor French-American-British (FAB) classification (eg, refractory anemia with
them with a growth advantage by decreasing theirrate of
excess blasts [RAEB], RAEB in transition, chronic myelodeath. These cellswould continue to cycleuntil further oncomonocytic leukemia). Mutations
of p53 have not been regenic events occurred.
ported in patients with either refractory anemia (RA) or RA
Acute lymphoblasticleukemia (ALL). We examined 330
with ring sideroblasts. (2) Many of the patients with a p53
samples of common-ALL, pre-pre B-ALL, and pre B-ALL
mutation havemonosomy of chromosome 17, similar to
and found a 2% to 3% incidence of p53 mutations in these
leukemias." Several smaller series found either a similar or
CML and AML.(3) Samples with p53 mutations have promslightly higher frequency of alterations of p53.47.1"1.1"2
These
inent p53 accumulation in their blast cells but not in their
p53 mutations appear mostoften in patientswithrelapse
mature and presumably MDS-derived myeloidcells,sugphase of
In contrast,type
(Burkitt's)
Lz
Bgesting that transcription of even mutated p53 is under norALL have a 50% incidence of p53 mutation^^^.^^' and often
mal control.
have activation of the c-myc gene. Although T-cell leukemia
Cases of AML and MDS have been reported that have
cell lines have about a 50% frequency of p53 mutations,""
high levels of p53 protein, as determined by either immunoprecipitation or immunochemistry, buthaveno detectable
p53 mutations are rarely found in patients with newly diagp53 m~tations.~"."' One
study found that the protein in AML
nosed T-ALL."" Interestingly, about 30% of samples from
patients with relapse T-ALL have p53mutations, suggesting
cells, although not mutated, often adopted the conformation
the importance of this alteration with progressive disease.
of mutant pS3 as identified by antibody pAb240 in AML.Xs
Patients with Li-Fraumeni syndrome (see below)have a 5 %
Additionally, p53 has been found in mutant conformation in
to 1 0 % frequency of childhood ALL, with the leukemic cells
normal human activated T lymphocytes and CD34' hematohaving a homozygous p53 mutations.
poietic stem cells.*' The wt-p53 is an allosteric protein that
Most of the p53 mutations in these hematopoietic maligcan exist potentially in two conformations that may be denanciesare G:Cto A:Ttransitionalnucleotidealterations
pendent on the cell cycle. This alteration of p53 conformaand most of these occur at CpG dinucleotides. This pattern
tion in normal dividing myeloid and lymphoid cells may be
of mutations probably results from spontaneousdeamination
associated with a temporary inability of the protein to halt
suggestingthat,in ALL and hemato~
cells at the G, phase allowing these cells to p r o l i f e r a t i ~ n . ~ ~at~ ~5-methylcytosine,
logic malignancies in general, the p53 mutations may not be
In AML, this "permanent" alteration in conformation could
induced by exogenous carcinogens. The codon location of
be a mechanism causing the preferential proliferation over
these mutations in any of the hematopoietic malignancies do
differentiation of AML cells. Related or independent is the
not appear toinfluence either the phenotype or clinical course
observation that about half of all AML sampleshave elevated
of these leukemias and lymphomas.
expression of mdm-2 (murine double minute).x' The human
Chronic lymphocytic leukerniu (CLL). The leukemic
homologue of mdm-2 gene may act in a negative feedback
cells have p53 mutations in about 15% of individuals with
loop with pS3. The mdm-2 gene was originally identified on
CLL." A minority of patients with CLL transform to a rapdouble-minutechromosomes thatwereamplifiedapproxiidly aggressive lymphoproliferative disorder that is known
mately %-fold in a spontaneously arisingtumorigenic Balb/c
as Richter's syndrome. At least 40% of these patients have
3T3 cellline") and overexpressionof this gene can transform
a p53 mutation in their more malignant cells." Therefore,
normal murine fibroblasts." The product of the mdm-2 gene
alterations of p53 are closely associated with transformation
binds towt-p53and
negativelyregulatestranscriptional
of CLL into a very aggressive lymphoma.
activation of ~ t - p 5 3 . ' ~ .It' ~can overcome wt-p53-mediated
Adult T-cell leukemia (ATL). About 30% to 50% of indisuppression of transformed cellular growth.'J In addition,
viduals with ATLhave p53 mutations in theirleukemic
wt-p53 can induce
mdm-2
expression," suggesting
an
cel]s.<l,.34.?5 In our series, leukemiccells of 4 of 10 acute
autoregulatory model for functionalactivity of p53."'."' Five
ATL cases had homozygous p53 mutations. In addition. we
percent to 30% of sarcomas have mdm-2 gene amplification
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
p53 IN HEMATOLOGIC MALIGNANCIES
studied an informative patient whose ATL cells had no detectable p53 mutation in the chronic phase of the disease. A
novel clone emerged that quickly expanded to acute ATL
and rapidly led to the death of the patient; these cells had a
homozygous missense mutation of ~ 5 3 The
. ~ data
~ suggest
that alterations of the p53 gene are frequently acquired abnormalities in acute ATL and may occur in the transition to
the more aggressive leukemic phenotype.
The natural history of development of acute ATL suggests
that HTLV-1 infection alone is not sufficient to cause leukemia. Many individuals infected with the virus do not develop
ATL; those who do often have a 20- to 30-year latency
period. HTLV-1 does not contain an oncogene, probably
does not activate proto-oncogenes by insertional activation,
nor does it inactivate tumor-suppressor genes by disrupting
them.'" A possible hypothesis concerning the development
of ATL can be stated as follows. HTLV-1 infection results
in acute expression of the tax product encoded from the X
region of HTLV-1, perhaps resulting in autocrine growth
stimulation through production of IL-2 and other lymphokines. 105.106 This stimulation may cause a polyclonal increase
of the HTLV-l-infected T lymphocytes. These cells may
periodically expand by various stimuli such as infections
that might enhance tax production. This polyclonally and
then oligoclonal expanded population of cells may have a
slight growth advantage over normal cells. Evolution from
chronic to acute ATL may result from additional mutations
such as a homozygous mutational alteration of p53. This
model is analogous to the progression of CML to blast crisis.
Leukemic lymphocytes staining positive for p53 are frequently found in patients with ATL even when the leukemic
cells have no p53 mutation. These cells are also Ki-67 positive, suggesting that the proliferating cells are staining positively for p53 protein. We examined normal, uninfected phytohemagglutinin (PHA)-stimulated T cells and HTLV-Itransformed T cells from the same normal individual and
found prominent p53 expression in both.Io7Further studies
are required to determine if the prominent expression of p53
in ATL cells represents either an aberrant p53 or a normally
expressed p53 in a rapidly proliferating population of T cells.
Similarities exist between HTLV and Epstein-Barr virus
(EBV). Both can easily immortalize lymphocytes in vitro.
Neither contain oncogenes nor transform by insertional mutagenesis. Most people infected with either of these viruses
are asymptomatic and only rarely develop a malignancy.
Infection withboth viruses can at least initially stimulate
polyclonal cellular proliferation. In both ATL and Burkitt's
lymphoma, p53 mutations are intimately associated with progression to malignancy. Burkitts lymphoma also is almost
always associated with activation of c-myc. Activation of an
oncogene is also likely to occur in ATL, butit has not
been identified as yet. Therefore, both viruses may provide
a growth advantage to a large cohort of cells. This active
proliferation over a long period of time may enhance the
opportunities of developing a p53 mutation.
Hodgkin ' S disease (HD). Immunohistochemical staining
for p53-protein on frozen- and paraffin-embedded lymph
node samples shows that a significant proportion of p53positive neoplastic cells are detected in about 60% to 80%
2415
of cases with mixed cellularity and nodular sclerosing type
~ ~ . 6 7 , 6 8 , 1 0 8I-O1 Immunoreactivity is localized to the nuclei of
Reed-Sternberg (RS) cells or its mononuclear variants. The
number of positive cells vary between 10%to 60%of recognizable RS cells. Mutations of p53 have been detected in
enriched RS cell preparations."
Accumulation of mutant
p53 only in RS cells suggests an important role of p53 in
the tumorigenic process of HD disease.
No correlation has been found between EBV infection
and p53 reactivity in RS cells.67The background of small
lymphocytes, plasma cells, eosinophils, and histiocytes in
the HD samples are unstained for p53. This finding is consistent with the view that RS cells (and variants) are the neoplastic components of HD. In addition, no p53 staining is
noted in lymphocyte-predominant HD. This finding is of
interest in view of the possibility that this disease may really
be a form of low-grade B-cell lymphoma rather than a subtype of HD.
Non-Hodgkin lymphoma (NHL). Aggressive, high-grade
B-cell NHL has about a 30% incidence of p53 mutations,
whereas indolent B-cell NHLrarely have alterations of
p53~68.70.71.113-115
About10% of T-cell NHL also have p53
mutations. We found that patients with acquired immunodeficiency syndrome (AIDS) have an increased incidence of
development of B-cell immunoblastic lymphomas (BIBL).
However, their incidence or type of p53 mutations does not
differ when compared with BIBL in individuals without human immunodeficiency virus (HIV) infe~tion.~'
In another
study of AIDS-related lymphoma, p53 was mutated only in
those samples histologically classified as small, noncleaved
cell lymphoma (SNCCL) and 63% of SNCCL samples had
p53 mutations."' Almost all of these samples hadc-myc
activation and were usually not infected with EBV.'16 The
c-myc is also dysregulated in Burkitt's lymphoma and L3type B-ALL, which also frequently have p53 alterations,
suggesting that those tissues with an abnormally expressed
c-myc protein may gain an additional growth advantage by
mutating the p53 gene.
Whereas 50% of aggressive B-cell NHL express high levels of p53, only 20% to 50% of these had demonstratable
p53 mutation^.^' Similarly, postthymic T-cell lymphomas
and CD30 (Ki-l)-positive, anaplastic large-cell lymphomas
frequently overexpress p53 in the absence of detectable p53
In contrast, cells of nonmalignant hyperplastic lymph nodes have negligible p53 expre~sion.~'
Therefore,
immunohistochemical analysis of p53 may help discriminate
between normal andneoplastic lymph nodes, but cannot distinguish wild-type from mutant p53 protein in this disease.
The nonmutated, but overexpressed, p53 may be stabilized
in these lymphoma cells by alternate mechanisms such as
binding to an additional protein, for example, mdm-2 or a
viral product.
Low-grade NHL rarely have p53 alterations, but their progression to high-grade lymphoma canbe associated with
development of p53 mutations. For example, serial biopsies
of patients with follicular NHL who underwent histologic
transformation showedthat one-third of the transformed
samples acquired a p53 mutation that was not detected in
the follicular stage of the disease."' Another study found
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
2416
IMAMURA, MIYOSHI. AND KOEFFLER
Table 3. Hematopoietic Malignancies in Which p53 Mutations Are
that 4 of 5 cases of transformation of follicular to diffuse
Associated With Disease Progression
large-cell NHL were associated with p53mutations.”’ Interestingly, in this study, one sample offollicular NHL had
1. Evolution from chronic phase to myeloid blast crisis ofCML.
regions of transformation to high-grade NHL; cells of this
2. Evolution from myelodysplastic syndrome to acute myelogenous
region,butnot
those of the follicularareas, contained a
leukemia.
mutant ~ 5 3 . ’ ”Particularly intriguing, p53-positive staining
3. Evolution from follicular to high-grade lymphoma.
4. Evolution from CLL to high-grade Richter’s-type lymphoma.
cellscanbedetectedbefore
transformation tohigh-grade
5. Progression t o a refractory phase of multiple myeloma.
lymphoma, but they represent a minority of the lymphoma6. Development of relapsed B- or T-ALL.
tous cells.”’ The percentage of p53 positive cells increased
either just before or after histologic progres~ion.’’~
Therefore, individuals with low-grade follicular NHL thathave
p53 staining cells may be at
increased risk for transformation
tions. p53 mutations have also been associated with progresto an aggressive NHL. These patients may be appropriate
sion of solid malignancies suchas the transition from benign
candidates to receive intensive therapy, but further studies
adenoma to malignantcolon carcinoma,”* evolution of glioare required to confirm this impression. The routine staining
m a ~ , ’ ~ ’ . and
’ ’ ~ development of metastatic prostate cancers.
of lymph nodes for p53 may become standard practice both
However, mutationsof p53havealso beenfoundin
the
for prognosis and therapy of lymphomas.
precancerous phases of adenocarcinomas, including adenoBurkitt’s lymphoma. We reported
that
Burkitt’s
a
matous polyps ofpatientswith
familialpolyposis coli,”’
lymphoma cell line (Raji) showed
increased amount of ~ 5 3 . ~suggesting
~
that p53 mutations may occur as an early event
Afterwards, several groups found that p53is usually mutated
in carcinogenesis as well.
Burkitt’s
in
lymphoma
More
cell
importantly,
Li-Fraumeni syndrome (LFS) and cancer families. LFS
the frequency of p53 mutations isabout35%to45%
in
isa rare autosomal dominantly inherited syndrome conBurkitt’s lymphomas from patient^.*^*^*' More than 50% of
sisting of the following clinicalcharacteristics:aproband
these mutations are clustered in a small stretch of 33 amino
with either acute lymphocytic leukemia, sarcoma,
breast canacids (codons 213 to 248), with codons 213 and 248 being
cer, brain tumor, andor adrenocortical carcinoma before the
the most frequently mutated spots; codon273, which is often
age of 45; a first-degree relative with a cancer in this group;
mutated in solid tumors, is rarely altered.”’ The p53 mutant
and a first- or second-degree relative with sarcoma at any
genesclonedfromthesecellshave
losttheirabilities
to
age or any cancer before age 45.’’‘ These individuals have
inhibit DNA synthesis, a characteristic of wt-p53.’”
a germline mutation of p53,which is consistent with the
Almost all cases of Burkitt’s have a chromosomal translofirst hit in Knudson’s two-hit mutational modelof hereditary
cancer.i37.13s
The developing cancers have
loss of the wtcation resulting in c-myc activation. Taken together with the
above presented SNCCL data, activation of myc and mutap53 allele (second hit) and retain the mutant p53 allele.’’9
tion of p53 may play a critical function in the development
Paradoxically, even though all the cells of these individuals
of a subgroup of B-cell lymphomas. The EBNA5 can bind
have a p53 mutation, they have a propensity to develop only
both RB and p53 and is required for B-cell transformation;
several types of cancer. Soft-tissue sarcomasusually develop
the role thatthisviralproteinplays
in pathogenesis of
in the first 5 years of life. Acute leukemias andbrain tumors
lymphomas requires further s t ~ d y . ~Clearly,
~ ~ , ~ more
~ ’ than
occur throughout childhood and young adulthood; the rare
one mechanism canlead to lymphomas, because, unlike Afadrenalcortical carcinomas occur primarilyininfancy. In
rican Burkitt’s, American Burkitt’s can often arise without
young adults, breast cancer is by far the most common neoEBV infection. The EBV-associated nasopharynegeal carciplasm. The incidence of ALL inindividuals with LFS is
nomas appear to have a normal ~ 5 3 . ~ * ’ . ’ * ~
about 6%.
Multiple myeloma. The p53 is mutatedin about 5% to
LFS is a rare genetic disorder. The prevalence of a germ10% of cases of multiple m y e l ~ m a . ’ ~ ~
Approximately
”*~
linep53 mutation is approximately 0.01% in the general
80% of human multiple myeloma cell lines have p53 mispopulation, 0.1% to 1% among various cancer patients, and
sense mutations.’” Little is known of p53 expression in the
5% to10% among young patientswith multiple cancer^.'^".^^'
plasma cells of patients with multiple mye10rna.l~’ Although
Roughly 50% of family members with LFS develop neoplanumbers are small, data suggest that development of a p53
sia by the ageof 30, in contrastto a 1% incidence of developmutation is a late event in the disease and is associated with
ment of cancer by the same cohortin the general population.
an aggressive c o ~ r s e . ~ ~ * , ~ * ~
Astriking feature of the affected members of families
p53 mutations and progression of hematopoietic maligwith LFS is their high frequency of second malignancies,
nancies. As shown on Table 3, p53 mutation is often assowith almost 50% of affected members developing morethan
ciatedwithprogressionof
the hematopoieticmalignancy.
one neoplasia. This observation has prompted several investigations of the incidence of germline p53 mutations in indiFor example, evolution of CML and MDS to myeloid blast
vidualswithmultiple cancers. In one study examining 59
crisis and AML, respectively, has been associated with loss
children and young adults who would not be considered as
of the short arm of chromosome 17 and mutationof the
having LFS, but who had developed two malignancies, the
remaining p53 allele. Also, several of the lymphoproliferaoverallfrequency of p53 mutationsintheir germline was
tive disorders, such as CLL and follicular lymphoma, can
approximately 7%.14’ One of the four tumors in this cohort
acquire a p53 alteration as they progress toa more malignant
was an NHL. In another study, 10 families whose relatives
phenotype. Relapsed T- and B-ALLoften develop p53muta-
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
2417
p53 IN HEMATOLOGIC MALIGNANCIES
had a high incidence of leukemias and lymphomas were
examined for p53 germline mutations and none was found.14'
In contrast, analysis for germline p53 mutations in individuals with childhood ALL containing p53 mutations in their
leukemic cells found that one (pre-B-ALL) of the four individuals had a germline alteration of p53.
Several observations of LFS can be made. (1) Germline
p53 mutations have been observed mostly in patients with
an unusual history of cancer, ie, either multiple malignancies
or family histones of cancers. (2) Most of the germline p53
mutations have been inherited; only 2 of 18 germline p53
mutations have been shown to occur de novo.'" In contrast,
germline retinoblastoma mutations most often (85%) occur
de novo in cases of retinoblastomas. (3) Although germline
p53 mutations can be widely distributed within the gene, an
increase predominance is found between codons 240 and
285. Almost all are missense point mutations. (4)Because
of the very low prevalence of germline p53 mutations in the
general population and the intensive analysis required to
detect germline p53 mutations, general screening of individuals is not indicated. An individual with cancer and a strong
family history of cancer or an individual with multiple neoplasias of the type occurring in LFS should be screened for
p53 mutations and, if positive, relatives at risk should then
be ~creened.'~'In addition, in the next few years, clinicians
must determine what type of surveillance should be given
to presymptomatic, germline carriers of p53 mutations.
Gene therapy and othertherapeuticapproaches.
Because of the pivotal role played by p53 in the regulation of
cell replication, the therapeutic restoration of expression of
wt-p53 in tumor cells having a p53 mutation might eventuallyplay a role in cancer gene therapy. In several model
systems, features of the tumor phenotype can be suppressed
in vitro through the restoration of expression of the mutant
tumor suppressor gene such as p53 and Rb. However, before
this method can serve as the basis for gene therapy of cancer,
many conceptual and technical problems must be solved.
Because such genetically modified cells will continue to contain and express other mutations, the mechanisms and frequency of reversion to the tumor phenotype must be examined. Experimentally, the transduced wt-p53 frequently
becomes mutated in transformed cells already expressing a
mutant ~ 5 3 . l Nevertheless,
~ ~ " ~ ~ colon adenocarcinoma cells
with other genetic defects in addition to the p53 gene became
nontumorigenic after being transfected with plasmids expressing a normal p53 allele.'46In the future, highly efficient
and targeted gene delivery vectors mustbe developed to
be clinically useful.'49 Experiments have shown that p53
retroviral vectors are capable of penetrating multiple cell
layers of three-dimensional tumor masses and mediating potential therapeutic effects.'" The regional administration of
viral supernatant containing p53 could be conceivably useful
therapeutically in established tumors and premalignant lesions of the upper aerodigestive tract and gastrointestinal
mucosa that had p53 mutations. At this time, none of the
delivery systems are powerful enough to be useful for in
vivo therapy. Furthermore, normal cells that are transduced
with the p53 expression vector may become dysfunctional
because of either inappropriate levels or timing of expression
of p53.
Immunotherapeutic approaches may also be possible. If
the mutant p53 epitopes were displayed on the malignant
cells and could be presented to T lymphocytes by class 1
major histocompatibility complex molecules on the plasma
membrane, a cytotoxic immune response might be selective
against these cancer cells. Indirect evidence suggests that
this may OCCUI-.'~' A murine model has shown that a cytotoxic
immune response can occur against cells expressing mutant
~53.'~
In' addition, autoantibodies to p53 have been identified in about 9% to 16% of cancer patients, including those
with breast cancer and lymphomas; no evidence exists that
these antibodies are directed against mutant p53 or that these
patients have an improved clinical response to their cancer,1s3-lss Although untested, the use of endogenously expressed mutant p53 as a target antigen remains a potential
approach for vaccine immunotherapy of selected cancers.
Identification of the genes that are regulated by p53 permits another future avenue of therapy. For example, p53
induces expression of a cyclin-dependent kinase inhibitor
known variously as either WAF-l, Picl, Cipl, SDI-l, or
p21, which helps regulate the cell cycle; cells with mutant
p53 lose this control. Perhaps therapeutically induced expression of WAF-l in p53 mutant cells will correct alterations in the cell cycle. For example, cells with p53 mutations do not stop at the GI phase of the cell cycle after DNA
damage. This contrasts to cells with wt-p53, which enforces
a pause at the G , phase of the cell cycle for DNA repair,
before beginning DNA synthesis. Finally, this difference in
cell cycle control after DNA damage between malignant
cells having mutant p53 and normal cells expressing wt-p53
can be used to design novel chemotherapy andor irradiation
treatment schedules.
In summary, p53 mutations occur moderately often in
hematopoietic malignancies. They are particularly associated
with progression of disease in both lymphoid and myeloid
leukemias as well as lymphomas. In addition, p53 mutations
occur very frequently in Burkitt's and other high-grade Bcell lymphomas. Also, only the RS cells in HD express high
levels of mutant p53, suggesting the major contribution of
these cells to this disease. Although mutant p53 maybe a
major cause of these malignancies, expression of this aberrant protein may also provide the handle for innovative approaches to these disease.
ACKNOWLEDGMENT
We thank Carl W. Miller, PhD, for helpful discussions and Kim
Burgin and Marge Jacobs for their excellent secretarial support.
REFERENCES
1. CulottaE,KoshlandDEJr:p53
Sweeps throughcancerresearch. Science 262:1958, 1993
2. Lane DP, Crawford LV: T antigen is bound to a host protein
inSV4O-transformed cells. Nature 278:261, 1979
3. Linzer DIH, Levine AJ: Characterizationof a 54K daltoncellular SV40 tumor antigen present in SV4O-transformedcells and uninfected embryonal carcinoma cells. Cell 17:43, 1979
4. Eliyahu D, Michalowitz D, Oren M: Overproduction of p53
antigen makes establishedcells highly tumorigenic. Nature 316:158,
1985
5. Parada LF, LandH, Weinberg RA, WolfD, Rotter V: Coopera-
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
2418
tion between gene encoding p53 tumor antigen and ras
in cellular
transformation. Nature 312:649, 1984
S: Immortalization of rat embryo fi6. Rovinski B, Benchimol
broblasts by the cellular p53 oncogene. Oncogene 2:445, 1988
S: P53:Oncogene or anti-oncogene'?
7. LaneDP,Benchimol
Genes Dev 4: I , 1990
8. Montenarh M: Biochemical properties of the growth suppressor/oncoprotein p53. Oncogene 7:1673, 1992
9. Vogelstein B, Kinzler KW: p53 function and dysfunction. Cell
70:523,1992
I O . Tuck SP, Crawford L: Overexpression of normal human p53
in established fibroblasts leads to their tumorigenic conversion. Oncogene Res 4:81, 1989
1 I . Eliyahu D, Raz A, Gruss P, Givol D, Oren M: Participation
of p53 cellular tumour antigen in transformation of normal embryonic cells. Nature 312546, 1984
12. JenkinsJR,RudgeK,CurrieGA:Cellularimmortalization
by a cDNA clone encoding the transformation-associated phosphoprotein p.53. Nature 312:651, 1984
13. Finlay CA, Hinds PW, Levine AJ: The p53 proto-oncogene
can act as a suppressor of transformation. Cell 57:1083, 1989
14. Eliyahu D, Michalovitz D, Eliyahu S, Pinhasi-Kimhi 0, Oren
M: Wild-type p53 can inhibit oncogene-mediated focus formation.
Proc Natl Acad Sci USA 86:8763, 1989
15. Hinds P, Finlay K, LevineAJ: Mutation is required to activate
the p53 gene for cooperation with therus oncogene and transformation. J Virol 63:739, 1989
16. Chen P-L, Chen YM, Bookstein R, Lee
W-H: Genetic mechanisms of tumorsuppressionbythehumanp53gene.Science
250: 1576, 1990
17. Levine AJ, Momand J, Finlay CA: The p53 tumour suppressor
gene. Nature 351:453, 1991
18. Finlay CA, Hinds PW, Tan TH, Eliyahu D, Oren M, Levine
AJ: Activating mutations for transformation by p53 produce a gene
product that forms an hsc70-pS3 complex with an altered half-life.
Mol Cell Biol 8:531, 1988
19. Wolf D, Rotter V: Major deletions in the gene encoding the
p53 tumor antigen cause lackof p53 expression in HL-60 cells. Proc
Natl Acad Sci USA 82:790, 1985
20. Mowat M, Cbeng AM, Kimura N, Bernstein A, Benchimol
S: Rearrangements of the cellular p53 gene
in erythroleukaemic cells
transformed by Friend virus. Nature 314:633, 1985
21.Masuda H, MillerC, Koeffler HP,Battifora H, Cline MJ:
Rearrangement of thep53geneiscommon
in humanosteogenic
sarcoma. Proc Natl Acad Sci USA 84:7716, 1987
22. Vogelstein B, Fearon ER, Kern SE, Hamilton SR, Preisinger
AC, Nakamura Y, White R: Allelotype
of colorectal carcinomas.
Science 244:207, 1989
23. Takahashi T, Nau MM, Chiba 1, Birrer MJ, Rosenberg RK,
VinocourM,LevittM,PassH,GazdarAF,Minna
JD: p53:A
frequenttargetforgeneticabnormalities
in lungcancer.Science
246:491,1989
24. Nigro JM, Baker SJ, Preisinger AC, Jessup JM, Hostetter R,
Cleary K, Bigner SH, Davidson N, Baylin S, Devilee P, Glover T,
Collins FS, Weston A, Modali R, Harris CC, Vogelstein B: Mutations in the p53 gene occur in diverse human tumour types. Nature
342:705,1989
25. Miller CW, AsloK, Tsay C, SlamonD, Ishizaki K, Toguchida
J, Yamamuro T, Lampkin B, Koeffler HP: Frequency and structure
Res 50:7950,
of p53 rearrangements in human osteosarcoma. Cancer
1990
26. Miller CW, Simon K,Aslo A, Kok K, Yokota I, Buys CHCM,
Terada M, KoefflerHP: p53 mutations in human lung tumors. Cancer
Res 52:1695, 1992
27. Gaidano G, Ballerini P, Gong JZ, Inghirami G, Neri A, New-
IMAMURA, MIYOSHI, AND KOEFFLER
comb EW, Magrath IT, Knowles DM, Dalla-FaveraR: p53 mutation
in human lymphoid malignancies associated with Burkit lymphoma
andchroniclymphocyticleukemia.Proc
Natl Acad Sci USA
88:5413, 1991
28. Hollstein MC, Metcalf RA, Welsh JA, Montesano R, Harris
CC: Frequent mutation of the p53 gene
in human esophageal cancer.
Proc Natl Acad Sci USA 87:9958, 1990
29. Hsu IC, Metcalf RA. Sun T, WelshJA,WangNJ,Harris
CC:Mutationalhotspotin
the p53gene in humanhepatocellular
carcinomas. Nature 350:427, 1991
30. Sidransky D, VoneschenbacjA,TsaiYC,JonesP,Summerhayes F, Marshall I, Paul M. Green P, Hamilton SR, Frost P,
Vogelstein B: Identification of p53 gene mutations
in bladder cancers
and urine samples. Science 252:706, 1991
31. Sakashita A, Hattori T, Miller CW, Suzushima H, Asou N,
Takatsuki K, Koeffler HP: Mutations of the p53 gene in adult Tcell leukemia. Blood 70:477, 1992
32.FaginJA,Matsuo
K, KarmakarA,ChenD-L,TangSH,
Koeffler HP: High prevalenceof mutations of the p53 genein poorly
1993
differentiated human thyroid carcinomas. J Clin Invest 91: 179,
33. Imamura J, Bartram CR, Berthold F, Harms D, Nakamura H,
KoeftlerHP: Mutation of the p53 gene in neuroblastomaandits
relationship with N - m y amplification. Cancer Res 53:4053, 1993
34. Sugito S, Yamato K, Sameshima Y, Yokota J, Yano S. Miyoshi I: Adult T-cell leukemia: Structures and expression of the p53
gene. Int J Cancer 49:880, 1991
35. Nagai H, Kinoshita T, Imamura J, Murakami Y, Hayashi K,
Mukai K, Ikeda S, Tobinai K, Saito H, Shimoyama M: Genetic
alteration of p53 in some patients with adult T-cell leukemia. Jpn J
Cancer Res 82: 1421, I991
36.Ichikawa A, Hotta T, Takagi N, TsushitaK,Kinoshita T,
Nagai H, Murakami Y, Hayashi K, Saito H: Mutations of p53 gene
and their relation to disease progression in B-cell lymphoma. Blood
79:2701, 1992
S, Cline MJ:
37.AhujaH, Bar-Eli M,AdvaniSH,Benchimol
Alterations in thep53geneandtheclonalevolution
of the blast
crisis of chronicmyelocyticleukemia.ProcNatl
Acad Sci USA
86:6783, 1989
38. Foti A, Bar-Eli M, Ahuja HG, Cline MJ: A splicing mutation
accounts for the lack of p53 gene expression in a CML blast crisis
cell line: A novel mechanism of p53 gene inactivation. Br J Haematol
76: 143, 1990
39. Ahuja H, Bar-Eli M, Arlin Z, Advani S, Allen SL, Goldman
J, Snyder D,Foti A, Cline M: The spectrumof molecular alterations
in theevolution of chronicmyelocyticleukemia.J
Clin Invest
87:2042, 199 1
40. Foti A, Ahuja HG, Allen SL, KoduruP, Schuster MW, Schulman P, Bar-Eli M, Cline MJ: Correlationbetweenmolecularand
clinical events in the evolution of chronic myelocytic leukemia to
blast crisis. Blood 77:2441, 1991
S, Kahn Y, Rechavi G,
41.Kelman 2, ProkocimerM,Peller
Manor Y, Cohen A, Rotter V: Rearrangements in the p53 gene in
Philadelphia chromosome positive chronic myelogenous leukemia.
Blood 74:23 18, 1989
42. Sugimoto K, Hirano N, Toyoshima H, Chiba S, Mano H,
Takaku F, Yazaki Y, Hirai H: Mutations of the p53 gene in myelodysplasticsyndrome(MDS)andMDS-derivedleukemia.Blood
8 1:3022, 1993
43. Sugimoto K, Toyoshima H, Sakai R, Miyagawa K, Hagiwara
K, Hirai H, lshikawa F, Takaku F: Mutations of the p53 gene in
lymphoid leukemia. Blood 77: 1 1.53, 1991
44. Sugimoto K, Toyoshima H, Sakai R, Miyagawa K, Hagiwara
K, Ishikawa F, Takaku F, Yazaki Y, Hirai H: Frequent mutations
in thep53gene
in humanmyeloidleukemiacelllines.Blood
79:2378,1992
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
p53 IN HEMATOLOGICMALIGNANCIES
45. Kuerbitz SJ, Plunkett BS, Walsh WV, Kastan MB: Wild-type
p53 is a cell cycle checkpoint determinant following irradiation.
Proc Natl Acad Sci USA 89:7491, 1992
46. Kastan MB, Zhan QM, El-Deiry WS, Carrier F, Jacks T,
Walsh WV, Plunkett BS, Vogelstein B, Fornace AJ Jr:A mammalian
cell cycle checkpoint pathway utilizing p53 and GADD45 is defective in ataxia-telangiectasia. Cell 71:587, 1992
47. Ponder B: Gene losses in human tumours. Nature 335:400,
1988
48. Harris N, Brill E, Shohat 0, Prokocimer M, Arai N, Wolf D,
Rotter V: Molecular basis for heterogeneity of the human p53 protein. Mol Cell Biol 6:4650, 1986
49. Orita M, Iwahana H, Kanazawa H, Hayashi K, Sekiya T:
Detection of polymorphisms of human DNA by gel electrophoresis
as single-strand conformation polymorphisms. Proc Natl Acad Sci
USA 862766, 1989
50. Orita M, Suzuki Y, Sekiya T, Hayashi K: Rapid and sensitive
detection of point mutations and DNA polymorphisms using the
polymerase chain reaction. Genomics 52374, 1989
51. Suzuki Y, Orita M, Shiraishi M, Hayashi K, Sekiya T: Detection of ras gene mutations in human lung cancers by single-strand
conformation polymorphism analysis of polymerase chain reaction
products. Oncogene 5:1037, 1990
52. Murakami Y, Hayashi K, Sekiya T: Detection of aberrations
of p53 alleles and the gene transcript in human tumor cell lines
by single-strand conformation polymorphism analysis. Cancer Res
51:3356, 1991
53. Hayashi K PCR-SSCP: A simple and sensitive method for
detection of mutations in the genomic DNA. PCR Methods Applications 1:34, 1991
54. Paquette RL, Karmakar A, Kizaki M, Wilcynzski SP, Miller
CW, Koeffler HP: Mutations of p53 and human papillomavirus infection occur independently in cervical carcinoma. Cancer 72: 1272,
1993
55. Hollstein M, Sidransky D, Vogelstein B, Harris CC: p53 mutations in human cancers. Science 253:49, 1991
56. Soussi T, Caron de Fromentel C, May P: Structural aspects
of the p53 protein in relation to gene evolution. Oncogene 5:945,
1990
57. Vogelstein B: A deadly inheritance. Nature 348:681, 1990
58. Borresen AL. Hovig E, Smith-Sorensen B, Malkin D, Lystad
S, Andersen TI, Nesland JM, Isselbacher KJ, Friend S: Constant
denaturant gel electrophoresis as a rapid screening technique for p53
mutations. Proc Natl Acad Sci USA 88:8405, 1991
59. Gibbs R, Caskey C T Identification and localization of mutations at the Lesch-Myhan locus by ribonuclease A cleavage. Science
236:303, 1987
60. Fischer SG, Lerman LS: DNA gragments differing by single
base-pair substitutions are separated in denaturing gradient gels:
Correspondence with melting theory. Proc Natl Acad Sci USA
80: 1579, 1983
61. Sheffield VC, Cox DR, Lerman LS, Myers RM: Attachment
of a 40-base-pair G + C-rich sequence (GC-clamp) to genomic DNA
fragments by the polymerase chain reaction results in improved
detection of single-base changes. Proc Natl Acad Sci USA 86932,
1989
62. Cotton RGH, Rodreques NR, Campbell RD: Reactivity of
cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of
mutations. Proc Natl Acad Sci USA 85:4397, 1988
63. Iggo R, Gatter K, BWek J, Lane D, Harris AL: Increased
expression of mutant forms of p53 oncogene in primary lung cancer.
Lancet 335:675, 1990
64. Gusterson BA, Anbazhagan R, Warren W, Midgley C, Lane
DP, O’Hare M, Stamps A, Carter R, Jayatilake H: Expression of
2419
p53 in premalignant and malignant squamous epithelium. Oncogene
6:1785, 1991
65. Cossman J, Schlegel R: p53 in the diagnosis of human neoplasia. J Natl Cancer Inst 83:980, 1991
66. Mashal R, Shtalrid M, Talpaz M, Kantarjian H, Smith L,
Beran M, Cork A, Trujillo J, Gutterman J, Deisseroth A: Rearrangement and expression of p53 in the chronic phase and blast
crisis of chronic myelogenous leukemia. Blood 75:180, 1990
67. Gupta RK, Norton A J , Thompson I W , Lister TA, Bodmer
JG: p53 expression in Reed-Stemberg cells of Hodgkin’s disease.
Leuk Suppl2:S31, 1993
68. Doglioni C, Pelosio P, Mombello A, Scarpa A, Chilosi M:
Immunohistochemical evidence of abnormal expression of the antioncogene-encoded p53 phosphoprotein in Hodgkin’s disease and
CD30+ anaplastic lymphomas. Hematol Pathol 5:67, 1991
69. Niedobitek G,Rowlands DC, Young LS, Herbst H, Williams
A, Hall P, Padfield J, Rooney N, Jones L Overexpression of p53
in Hodgkin’s disease: Lack of correlation with Epstein-Barr virus
infection. J Pathol 169:207, 1993
70. Villuendas R, Piris MA, Orradre JL, Mollejo M, Algara P,
Sanchez L, Martinez JC, Martinez P: P53 protein expression in
lymphomas and reactive lymphoid tissue. J Pathol 166:235, 1992
7 1. Nakamura H, Said J W , Miller CW, Koeffler H P Mutation and
protein expression of p53 in acquired immunodeficiency syndromerelated lymphomas. Blood 82920, 1993
72. Gannon JV, Greaves R, Iggo R, Lane DP: Activating mutations in p53 produce a common conformational effect. A monoclonal
antibody specific for the mutant form. EMBO J 9:1592, 1990
73. Lubbert M, Miller CW, Crawford L, Koeffler HP: p53 in
chronic myelogenous leukemia. Study of mechanisms of differential
expression. J Exp Med 167:873, 1988
74. Nakai H, Misawa S, Toguchida J, Yandel DW, Ishizaki K
Frequent p53 gene mutations in blast crisis of chronic myelogenous
leukemia, especially in myeloid crisis harboring loss of a chromosome 17p. Cancer Res 526588, 1992
75. Feinstein E, Gale RP, Reed J, Canaani E: Expression of the
normal p53 gene induces differentiation of K562 cells. Oncogene
7:1853, 1992
76. Koeffler HP, Miller C, Nicolson MA, Ranyard J, Bosselman
RA: Increased expression of p53 protein in human leukemia cells.
Proc Natl Acad Sci USA 83:4035, 1986
77. Smith LJ, McCulloch EA, Benchimol S. Expression of the
p53 oncogene in acute myeloblastic leukemia. J Exp Med 164:751,
1986
78. Prokocimer M, Shaklai M, Bassat HB, Wolf D, Goldfinger
N, Rotter V: Expression of p53 in human leukemia and lymphoma.
Blood 68:113, 1986
79. Slingerland JM, Minden MD, Benchimol S: Mutation of the
p53 gene in human acute myelogenous leukemia. Blood 77:1500,
1991
80. Fenaux P, Jonveaux P, Quiquandon I, Lai JL, Pignon JM,
Loucheux-Lefebvre MH, Bauters F, Kerckaert J P : P53 gene mutations in myeloid leukemia with 17p monosomy. Blood 78:1652,
1991
81. Fenaux P, Preudhome C, Quiquandon I, Jonveaux P, Lai
JL, Vanrumbeke M, Loucheux-Lefebvre MH, Bauters F, Berger R,
Kerckaert JP: Mutations of the p53 gene in acute myeloid leukaemia.
Br J Haematol 80:178, 1992
82. Wada M, Bartram CR, Nakamura H, Hachiya M, Chen DL,
Borenstein J, Hansen-Hagge TE, Ludwig WD, Reiter A, Mizoguchi
H, Koeffler HP. Analysis of p53 mutations in large series of
lymphoid hematological malignancies of childhood. Blood 823163,
1993
83. Preudhomme C, Quesnel B, Vachee A, Lepelley P, CollynD’Hooghe M, Wattel E, Fenaux P: Absence of amplification of
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
2420
MDM2 gene, a regulator of p53 function, in myelodysplastic syndromes. Leukemia 7: 129 I . 1993
84. Jonveaux PH, Fenaux P. Quiquandon I, Pignon JM, Lai JL.
Loucheux-Lefebvre MH, Goossens M, Bauters F, Berger R: Mutationsinthep53gene
in myelodysplasticsyndromes.Oncogene
6:2243, 199 1
85. Zhang W, Hu Guiying, Estey E, Hester J, Deisseroth A: Altered conformation of the p53 proteinin myeloid leukemia cells and
mitogen-stimulated normal blood cells. Oncogene 7: 1645, 1992
86. Rivas CI, Wisntewski D, Strife A, Perez A, Lambek C, Bruno
S, DarzynkiewiczZ,ClarksonB:Constitutiveexpressionofp53
protein in enrichednormalhumanmarrow
blast cell populations.
Blood 79:1982, 1992
87. Milner J, Medcalf EA: Cotranslation of activated mutant p53
with wild typedrivesthewild-typep53
protein intothemutant
conformation. Cell 65:765, I99 1
88. Milner J: Differentforms of p53detected by monoclonal
antibodies in non-dividing
and
dividing
lymphocytes.
Nature
310:143,1984
89.Bueso-RamosCE,Yang
Y, deLeon E, McCown P, Stass
SA, Albitar M: The human MDM-2 oncogene is overexpressed in
leukemias. Blood 82:2617, 1993
90. Cahilly-Snyder L, Yang-Feng T, Francke U, George DL: Molecular analysis and chromosomal mapping of amplified genes isolated fromatransformedmouse3T3
cell line. SomaticCellMol
Genet 13:235, 1987
91. Fakharzadeh SS, Trusko SP, George DL: Tumorigenic potential associated with enhanced expression of a gene that is amplified
in a mouse tumor cell line. EMBO J 10:1565, 1991
92. Barak Y, Oren M: Enhanced binding of a 95 kDa protein to
p53 in cellsundergoingp53-mediatedgrowtharrest.EMBOJ
I1:21 15. 1992
93. Momand J, Zambetti GP, Olson DC, George DL, Levine AJ:
The mdm-2 oncogene product forms a complex with the p53 protein
and inhibits p53 mediated transactivation. Cell 69:1237, 1992
94.FinlayCA: The mdm-2oncogenecanovercomewild-type
p53 suppression of transformed cell growth. Mol Cell Biol 13:301,
1993
95. Barak Y , Juven T, Haffner R, Oren M: mdm2 expression is
induced by wild type p53 activity. EMBO J 12:461, 1993
96. Zdmbetti GP,Levine AJ. Acomparison of thebiological
activities of wild-type and mutant p53. FASEB J 7:855, 1993
97. Oliner JD, Kinzler KW, Meltzer PS, George DL, Vogelstein
B: Amplification of a geneencoding a p53-associatedproteinin
human sarcomas. Nature 358:80, 1992
98. Yonish G, Resnitzky D, Lotem J, Sachs L, Kimcbi A, Oren
M: Wild-type p53 induces apoptosis myeloid leukaemic cells that
is inhibited by interleukin-6. Nature 352:345, 1993
99. Low SW,Schmitt EM, Smith SW, Osborne BA, Jacks T: p53
is requiredforradiation-inducedapoptosis
in mousethymocytes.
Nature 362:847, 1993
100. Clark AR, Purdie CA, Harrison DJ. Morris RC, Bird CC,
HooperML,WylieAH:Thyomocyteapoptosisinduced
by p53dependent and independent pathways. Nature 363:849, 1993
I O I , Fenaux P, Jonveaux P, Quiquandon I, Preudhomme C, Lai
JL, Vanrumbeke M, Loucheux-Lefebvre MH, Bauters F, Berger R,
KerckaertJP:Mutations of thep53gene in B-cell lymphoblastic
acute leukemia: A report on 60 cases. Leukemia 6:42, 1992
102. Felix CA, Nau MM,Takasashi T, Mitsudomi T, Chiba 1,
Poplak DC, Reaman CH,ColeDE,LetterioJJ,Whang-PengJ,
Knutsen T, Minna JD: Hereditary and acquired p53 gene mutations
in childhoodacutelymphoblasticleukemia.JClinInvest89:640,
I992
103. Cheng J, Haas M: Frequentmutationsinthep53tumor
IMAMURA,MIYOSHI, AND KOEFFLER
suppressorgene in humanleukemiaT-celllines.
Mol Cell Biol
I0:5502.1990
104. Norley Sg, Kurth R: Retroviruses and malignant lymphoma.
Blut 58:221, 1989
105. Seiki M, Hattori S, Hirayama Y, Yoshida M: Human adult
T-cell leukemia virus: Complete nucleotide sequence of the provirus
genome integrated in leukemia cell DNA. Proc Natl Acad Sci USA
80:3618, 1983
106. Sodroski J, Rosen C, Goh WC, Haseltine W:
A transcriptional activator protein enccoded by the x-lor region of the human
T-cell leukemia virus. Science 228:1430, 1985
107. Lubbert M, Miller CW, Kahan J, Koeffler HP: Expression,
methylationandchromatinstructure
of thep53geneinuntransformed and human T-cell leukemia virus type I-transformed human
T-lymphocytes. Oncogene 4:643, 1989
108. SaidJW,BarreraR,ShinatakuIP,Nakamura
H, Koeffler
HP: Immunohistochemical analysis of p53 expression in malignant
lymphomas. J Pathol 141:1343, 1992
109. Doglioni C, Pelosio P, Mombello A, Scarpa A, Chilosi M:
Immunohistochemical evidence for abnormal expression of the antiin Hodgkin’sdiseaseand
oncogene-encodedp53phosphoprotein
CD30+ anaplastic lymphomas. Hematol Pathol 5:67,
1991
1 IO. Villuendas R, Piris MA, Orradre JL, Mollejo M, Algara P,
SanchezL,MartinezJC,Martinez
P: p53proteinexpression
in
lymphomas and reactive lymphoid tissue. J Pathol 166:235, 1992
11 I . Gupta RK, Patel K, Bodmer WF, Bodmer JG: Mutation
of
p53 in primary biopsy material and cell lines from Hodgkin’s disease. Proc Natl Acad Sci USA 90:2817, 1993
I 12. Trumper LH, Brady G, Bagg A, Gray D, Loke SL, Griesser
S, IscoveNN,
H, WagmanR,Braziel
R, GascoyneRD,Vicini
Cossmann J, Mak TW: Single-cell analysis of Hodgkin and ReedSternberg cells: Molecular heterogeneity of gene expression and p53
mutations’. Blood 81:3097, 1993
I 13. Lo Coco F, Gaidano G, Louie DC, Offit K, Chaganti RSK,
Dalla-Favera R: p53 mutations are associated with histologic transformation of follicular lymphoma. Blood 82:2289, 1993
114. Farrugia M, Duan LJ, Reis MD, Ngan BY, Berinstein NL:
Alterations of the p53 tumor suppressor gene in diffuse large cell
lymphomas with translocations of the c-myc and BCL-2 proto-oncogenes. Blood 83:191, 1994
I 15. RodriquezMA,Ford
RJ, Goodacre A, SelvanayagamP,
Cabanillas F, Deisseroth AB: Chromosome 17- and p53 changes in
lymphoma. Br J Haematol 79575, 1991
V, Saglio G,
116. Ballerini P, Gaidano G,GongJZ,Tassi
Knowles DM, Dalla-Favera R: Multiple genetic lesions in acquired
immunodeficiency
syndrome-related
non-Hodgkin’s
lymphoma.
Blood 8 1 : 166, 1993
I 17. Cesarman E, Inghirami G, Chadburn A, Knowles DM: High
levels of p53 proteinexpression do notcorrelatewithp53gene
mutations in CD30 (Ki-l) positive anaplastic large cell lymphoma.
Modern Pathol 6:87A, 1993
118. Matsushima A, Cesarman A, Chadbum A, Liu J, Knowles
DM: p53proteinoverexpressionfrequentlyoccursbutp53gene
mutations infrequently occurin post thymic T cell lymphomas. Modem Pathol 6:95A, 1993
119. Sander CA, Yano T, Clark HM, Harris C, Longo DL, Jaffe
ES. Raffeld M: p53 mutation is associated with progressionin follicular lymphomas. Blood 82:1994, 1993
120. Fanell PJ, Allan GJ, Shanahan F, Vousden KH, Crook T:
p53 is frequently mutated in Burkitt’s lymphoma cell lines. EMBO
J 10:2879, 1991
I2 I . Wiman KG, Magnusson W, Ramqvist T, Klein G: Mutant
p53 detected in a majority of Burkitt lymphoma cell linesby monoclonal antibody PAb240. Oncogene 6:1633, 1991
122. Vousden KH, Crook T, Farrell PJ: Biological activities of
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
p53 IN HEMATOLOGICMALIGNANCIES
p53 mutants in Burkitt’s lymphoma cells. J General Virol 74:803,
1993
123. Bhatia KS, Gutie’rrez MI, Huppi K, Siwarski D, Magrath
IT: The pattern of p53 mutations in Burkitt’s lymphoma differs from
that of solid tumors. Cancer Res 52:4273, 1992
124. Mannick JB, Cohen JI, Birkenbach M, Marchini A, Kieff
E: The Epstein-Barr virus nuclear protein encoded by the leader of
the EBNA RNAs is important in B-lymphocyte transformation. J
Virol 65:6826, 1991
125. Effert P, McCoy R, Abdel-Hamid M, Fiynn K, Zhang Q,
Busson P, Tursz T, Liu E, Raab-Traub N: Alterations of the p53
gene in nasopharyngeal carcinoma. J Virol 66:3768, 1992
126. Spruck Ch, Tsai YC, Huang DP, Yang AS, Rideout WM
111, Gonzales-Zulueta M,Choi P, Lo K-W, Yu MC, Jones PA:
Absence of p53 gene mutations in primary nasopharyngeal carcinomas. Cancer Res 52:4787, 1992
127. Preudhomme C, Facon T, Zandecki M, Vantumbeke M, Lai
JL, Nataf E, Loucheux-Lefebvre MH, Kerckaert JP, Fenaux P Rare
occurrence of p53 gene mutations in multiple myeloma. Br J Haemato1 81:440, 1992
128. Portier M, Moles JP, Mazars GR, Jeanteur Ph., Bataille R,
Klein B, Theillet Ch: p53 and RAS gene mutations in multiple
myeloma. Oncogene 7:2539, 1992
129. NerA, Baldini L, Trecca D, Cro L, Polli E, Maiolo A T
p53 gene mutations in multiple myeloma are associated with advanced forms of malignancy. Blood 81: 128, 1993
130. Mazars GR, Portier M, Zhang XG,Jourdan M, Bataille R,
Theillet Ch. Klein B: Mutations of the p53 gene in human myeloma
cell lines. Oncogen 7:1015, 1992
131. Palumbo AP, Pileri A, Dianzani U, Massaia M, Boccadoro
M, Calabretta B: Altered expression of growth-regulated protooncogenes in human malignant plasma cells. Cancer Res 49:4701, 1989
132. Baker SJ, Fearon ER, Nigro JM, Hamilton SR, Preisinger
AC, Jessup JM, van Tuinen P, Ledbetter DH, Barker DF, Nakemura
Y, White R, Vogelstein B: Chromosome 17 deletions and p53 gene
mutations in colorectal carcinomas. Science 244:217, 1989
133. Sidransky D, Mikkelsen T, Schwechheimer K, Rosenblum
ML, Cavanee W, Vogelstein B: Clonal expansion of p53 mutant
cells is associated with brain tumor progression. Nature 355:846,
1992
134. Fults D, Brockmeyer D, Tullous MW, Pedone CA, Cawthon
RM: p53 mutations and loss of heterozygosity on chromosome 17
and 10 during human astrocytoma progression. Cancer Res 52:674,
1992
135. Shirasawa S, Urabe K, Yanagawa Y, Toshitani K, Iwama
T, Sasazuki T: p53 gene mutations in colorectal tumors from patients
with familial polyposis coli. Cancer Res 51:2874, 1991
136. Garber JE, Goldstein AM, Kantor AF, Dreyfus MG,
Fraumeni JF, Li FP: Follow-up study of twenty-four families with
Li-Fraumeni syndrome. Cancer Res 51:6094, 1991
137. Malkin D, Li FP, Strong LC, Graumeni JF, Nelson CE, Kim
DH, Kassel J, Gryka MA, Bischoff FZ, Tainsky MA, Friend SH:
Germline p53 mutations in a familial syndrome of breast cancer,
sarcomas, and other neoplasms. Science 250:1233, 1990
138. Srivastava S, Zou ZQ, Pirollo K, Blattner W, Chang EH:
Germ-line transmission of mutated p53 gene in a cancer-prone family with Li-Fraumeni syndrome. Nature 348:747, 1990
139. Li FP, Prraumeni JF, Mulvihill JJ, Blattner WA, Dreyfus
2421
MG, Tucker MA, Miller RW: A cancer family syndrome in twentyfour kindreds. Cancer Res 48:558, 1988
140. Li FP, Correa P, Fraumeni JF: Testing for germ line p53
mutations in cancer families. Cancer Epidemiol Biomarkers Prev
1:91, 1991
141. Toguchida J, Yamaguchi T, Herrera G, Beauchamp R, Yandell DW: A survey of germ-line and somatic p53 gene mutations in
patients with bone and soft tissue sarcomas. Am J Hum Genet
49:(S)458, 1991
142. Malkin D, Jolly KW, Barbier N, Look AT, Friend SH, Gehhardt MC, Anderson TI, Borrensen A-L, Li FB, Garber J, Strong
LC: Germline mutations of the p53 tumor-suppressor gene in children and young adults with second malignant neoplasms. N Engl J
Med 326: 1309, 1992
143. Felix CA, D’Amico D, Mitsudomi T, NauMM, Li FP,
Fraumeni JP Jr, Cole DE, McCalla J, Reaman GH, Whang-Peng J:
Absence of hereditary p53 mutations in 10 familial leukemia pedigrees. J Clin Invest 90:653, 1992
144. Frebourg T, Friend SH: Cancer risks from germline p53
mutations. J Clin Invest 9 0 1637, 1992
145. Li FP, Garber JE, Friend SH, Strong LC, Patenaude AF,
Juengst ET, Reilly PR, Coma P, Fraumeni JF: Recommendations
on predictive testing for germline p53 mutations among cancer-prone
individuals. J Nat Cancer Inst 84:1156, 1992
146. Baker SJ, Markowitz S, Fearon ER, Willson JKV, Vogelstein B: Suppression of human colorectal carcinoma cell growth
by wild-type p53. Science 249:912, 1990
147. Diller L, Kassel J, Nelson C, Gryka MA, Litwak G, Gebhardt
M, Bressac B, Ozturk M, Baker SJ, Vogelstein B, Friend SH: p53
functions as a cell cycle control protein in osteosarcomas. Mol Cell
Biol 10:5772, 1990
148. Johnson P, Gray D, Mowat M, Benchimol S: Expression of
wild-type p53 is not compatible with continued growth of p53negative tumor cells. Mol Cell Biol 11 :1, 1991
149. Friedmann T: Gene therapy of cancer through restoration of
tumor-suppressor functions? Cancer 70: 1810, 1992
150. Fujiwara T, Grinmm EA, Mukhopadhyay T, Cai DW, OwenSchaub LB, Roth JA: A retroviral wild-type p53 expression vector
penetrates human lung cancer spheroids and inhibits growth by inducing apoptosis. Cancer Res 53:4129, 1993
151. Wiedenfeld EA, Fernandez-Vina M, Berzofsky JA, Carbone
DP: Evidence for selection against human lung cancers bearing p53
missense mutations which occur withintheHLA A*0201 peptide
consensus motif. Cancer Res 54:1175, 1994
152. Yanuck M, Carbone DP, Pendleton CD, Tsukui T, Winter
SF, Minna JD, Berzofsky JA: A mutant p53 tumor suppressor protein
is a target for peptide-induced CD8’ cytotoxic T-cells. Cancer Res
53:3257, 1993
153. Caron de Fromentel C., May-Levin F, Mouriesse H, Lemerle
J, Chandrasekaran K, MayP: Presence of circulating antibodies
against cellular protein p53 in a notable proportion of children with
B-cell lymphoma. Int J Cancer 39:185, 1987
154. Crowford LV, Pim DC, Bulbrook RD: Detection of antibodies against the host protein p53 in sera from patients with breast
cancer. Int J Cancer 30:403, 1982
155. Winter Sf, Minna JD, Johnson BE, Takahashi T, Gazdar
AF, Carbone D P Development of antibodies against p53 in lung
cancer patients appears to be dependent on the type of mutation.
Cancer Res 52:4168, 1992
From www.bloodjournal.org by guest on June 17, 2017. For personal use only.
1994 84: 2412-2421
p53 in hematologic malignancies
J Imamura, I Miyoshi and HP Koeffler
Updated information and services can be found at:
http://www.bloodjournal.org/content/84/8/2412.citation.full.html
Articles on similar topics can be found in the following Blood collections
Information about reproducing this article in parts or in its entirety may be found online at:
http://www.bloodjournal.org/site/misc/rights.xhtml#repub_requests
Information about ordering reprints may be found online at:
http://www.bloodjournal.org/site/misc/rights.xhtml#reprints
Information about subscriptions and ASH membership may be found online at:
http://www.bloodjournal.org/site/subscriptions/index.xhtml
Blood (print ISSN 0006-4971, online ISSN 1528-0020), is published weekly by the American
Society of Hematology, 2021 L St, NW, Suite 900, Washington DC 20036.
Copyright 2011 by The American Society of Hematology; all rights reserved.