Download Identification of novel target proteins in sebaceous gland carcinoma

ORIGINAL ARTICLE
Identification of novel target proteins in sebaceous gland carcinoma
Boban M. Erovic, MD,1 Ayman Al Habeeb, MBBS,2 Luke Harris, MD,1 David P. Goldstein, MSc, MD,1 Dae Kim, MBChB, PhD,1 Danny Ghazarian, MD,2
Jonathan C. Irish, MSc, MD1*
1
Department of Otolaryngology–Head and Neck Surgery/Surgical Oncology, Princess Margaret Hospital, University of Toronto, Toronto, Canada, 2Department of Laboratory
Medicine and Pathobiology, Princess Margaret Hospital, University of Toronto, Toronto, Canada.
Accepted 29 February 2012
Published online 19 June 2012 in Wiley Online Library (wileyonlinelibrary.com). DOI 10.1002/hed.23021
ABSTRACT: Background. The aim of this study was to identify new
target proteins in sebaceous gland carcinoma.
Methods. A tissue microarray containing 115 core biopsies was
constructed and stained for proteins involved in carcinogenesis,
angiogenesis, inflammation, and cell-to-cell contact. Two investigators
independently determined protein expression of all antibodies.
Results. Vascular endothelial growth factor receptor 2 (VEGFR-2),
platelet-derived growth factor receptor alpha and beta (PDGFR-a/-b),
epidermal growth factor receptor (EGFR), cyclooxygenase 1 and 2
(Cox-1/-2), myeloid cell leukemia sequence 1 (Mcl-1), matrix
metalloproteinase 1 (MMP-1), CD9, Bmi-1, 14-3-3r, glutathione S-
INTRODUCTION
Sebaceous gland carcinomas are often aggressive skin
malignancies arising most commonly from the meibomian
or Zeis glands.1 There is a high tendency for metastatic
disease particularly to regional lymph nodes.2 Unfortunately, despite early and aggressive surgical treatment,
patients with sebaceous gland carcinomas have a high
incidence rate of locoregional recurrence and distant metastatic disease.2 Epidemiological studies have shown that
during the past 30 years there has been little change in
the stage at which the disease presents, and little
improvement in the prognosis for patients with sebaceous
gland carcinoma.1
Given the high recurrence rate despite aggressive surgical treatment, an effective systemic adjuvant therapy
would represent a major breakthrough in the management
of these patients. To the best of our knowledge there are
no studies that have looked into novel targeted systemic
therapies in the treatment of sebaceous gland carcinoma.
In the current study we aimed to identify proteins that
are overexpressed in sebaceous gland carcinoma cells,
with the hope that they may be amenable to therapeutic
targeting. We examined the expression of specific proteins involved in angiogenesis, inflammation, apoptosis,
cell proliferation, cell-to-cell contact, and carcinogenesis.
*Corresponding author: J. C. Irish, Princess Margaret Hospital, Wharton Head
and Neck Centre, 610 University Avenue, 3rd Floor, Toronto, ON M5G 2M9,
Canada. E-mail: [email protected]
642
HEAD & NECK—DOI 10.1002/HED
MAY 2013
transferase pi (Gstp), and members of the sonic hedgehog (SHH), AKT,
and WNT pathways were significantly overexpressed in sebaceous gland
carcinomas.
Conclusions. We have demonstrated for the first time that proteins related
to angiogenesis, inflammation, and cell proliferation are overexpressed in
sebaceous gland carcinomas. These proteins may hold promise as
novel therapeutic targets for the treatment of sebaceous gland carcinoma.
C 2012 Wiley Periodicals, Inc. Head Neck 35: 642–648, 2013
V
KEY WORDS: target proteins, sonic hedgehog, angiogenesis,
targeted anticancer therapy
This was accomplished through the use of a high-throughput tissue microarray containing 35 tumor samples from
20 patients with sebaceous gland carcinoma. Tissue
microarrays underwent immunohistochemistry staining
and 2 investigators independently determined protein
expression.
MATERIALS AND METHODS
Patients
Patients with sebaceous gland carcinoma managed at
the Princess Margaret Hospital (PMH) between 1983 and
2009 were identified through the cancer registry. Patients
were included if they had a newly diagnosed and previously untreated sebaceous gland carcinoma that was managed at the PMH. Sufficient paraffin-embedded tissue to
perform the microarray was an additional criterion for
inclusion. Patients with incomplete medical records and
those who did not pursue curative treatment were
excluded. Approval was obtained from the institutional
Research Ethics Board. The extent of disease was classified using the tumor-node-metastasis (TNM) staging system, in accord with the American Joint Committee on
Cancer (AJCC) classification. The tumors were classified
as periorbital if they occurred within the orbit or on the
eyelids and were classified as extraorbital if they arose
from another head and neck site.
At the PMH, the primary management approach for
patients with sebaceous gland carcinoma is primary surgery, with adjuvant radiotherapy offered to those patients
with positive margins or nodal metastases. The University
TARGET
PROTEINS IN SEBACEOUS GLAND CARCINOMAS
TABLE 1. Antibodies, sources, and retrieval buffers for each antibody used in this study.
Antibody
Source
Catalog no.
Clone
Pretreatment
Dilution (Incubation)
14-3-3Sigma
Bcl-2 alpha
Bcl-xl AB-2
Bmi-1
Catenin beta
Cdc2p34
CD9
Cytokeratin 7
EGFR
Gli-1
Gli-2
Gli-3
Mcl-1
MMP 1
PDGFR alpha
PDGFR beta
SHH
Smo
VEGFR2
WISP-1
WISP-2
Gst-p
pAKT
pmTOR ser2448
FOX-01
Chromogranin
p27
p21
p53
Cyclin D1
Rb
Cox-1
Cox-2
Thermo Scientific/Neomarkers
Thermo Scientific/Neomarkers
Thermo Scientific/Neomarkers
Millipore
Santa Cruz
Santa Cruz
Novocastra
Thermo Scientific/Neomarkers
Invitrogen
Santa Cruz
Aviva
Santa Cruz
Neomarkers
Lab Vision/Neomarkers
abcam
Thermo Scientific/Neomarkers
Santa Cruz
Santa Cruz
Santa Cruz
Santa Cruz
Santa Cruz
Novocastra
Cell Signaling
Cell Signaling
Novus Biological
Neomarkers
Transduction Lab
Pharmingen
Novocastra
Lab Vision
Pharmingen
Santa Cruz (Goat)
Santa Cruz (Goat)
MS-1185-PO
MS-123-PO
MS-1334-PO
05-637
Sc-7963
Sc-54
NCL-CD9
MS-1352-PO
28-0005
Sc-20687
ARP31885_T100
Sc-20688
MS-681-PO
RB-9225-PO
Ab61219
RB-1692-PO
Sc-9024
Sc-13943
Sc-6251
Sc-25441
Sc-25442
NCL-GSTpi-438
3787S
4976S
NBPI-19825
RB9003-PO
610242
556431
NCL p53-DO7
RM 9104
554136
Sc-1754
Sc-1747
1433S01
100/D5
7D9
F6
TE9
TE9
TE9
TE9
Citrate
Citrate
TE9
Trypsin
Pepsin
Citrate
TE9
TE9
Trypsin
Citrate
TE9
TE9
Citrate
Trypsin
TE9
Citrate
Trypsin
Citrate
Citrate
Citrate
Citrate
Pepsin
Citrate
TE9
Citrate
TE9
Citrate
TE9
Citrate
1/400
1/100
1/1000
1/400 overnight
1/50 overnight
1/1000 overnight
1/200 1 hour
1/300 overnight
1/100 1 hour
1/100 overnight
1/100 overnight
1/100 overnight
1/100 overnight
1/800 overnight
1/300 overnight
1/100 overnight
1/400 overnight
1/300 overnight
1/1000 1 hour
1/100 overnight
1/500 overnight
1/300 1 hour
1/50 overnight
1/50 overnight
1/100 overnight
1/500 1 hour
1/1000 1 hour
1/200 overnight
1/300 1 hour
1/200 overnight
1/300 1 hour
1/100 overnight
1/1000 overnight
Health Network skin pathology site group (D. Ghazarian
and A. Al Habeeb) reviewed all sebaceous gland carcinoma cases.
Statistical analysis
Statistical analysis was performed using the Statistical
Package for the Social Sciences software (SPSS version
15.0; SPSS Inc., Chicago, IL). Demographic and pathologic data were summarized using descriptive statistics.
The recurrence and survival rates were calculated from
the date of diagnosis to the event of interest. Recurrences
were classified as local, regional, or distant. Survival
analysis was performed using the Kaplan–Meier method.
The log-rank method was used for univariate analysis.
All the statistical tests were 2-tailed, and a value of p <
.05 was considered significant.
Specimens
A total of 35 surgical specimens from 20 patients contained sufficient tissue to be sourced for construction of a
tissue microarray (TMA). Two skin pathologists reviewed
all hematoxylin-eosin–stained tumors.
72F6
OV/TL 12/30
31G7
Polyclon
Polyclon
Polyclon
RC13
Polyclon
Polyclon
Polyclon
Polyclon
Polyclon
Polyclon
LW29
49F9
Tissue microarray construction
After examination of all available slides, the most representative blocks were chosen from each of the 35 specimens. TMAs were constructed with 3 cores from each
specimen. Each core measured 0.6 mm in diameter. The
cores were mounted on linear arrays in a single paraffin
TMA block. A character map was constructed delineating
the various specimens and their order in the TMA. Tumor
samples were categorized as negative, weak, moderate, or
high when 0% to 10%, 11% to 30%, 31% to 60%, and
61% to 100% of tumor cells stained positively, respectively. Again, 2 investigators (A.A. and D.G.) independently determined the immunohistochemical expression of
all antibodies.
Immunohistochemistry staining
All details regarding the antibody, source, clone, pretreatment, and dilution are summarized in Table 1. Formalin-fixed paraffin-embedded sections (4 lm) were dewaxed
in 5 changes of xylene and brought down to water through
graded alcohols. If necessary, antigen retrieval and
unmasking procedures were applied (see the following
text). Endogenous peroxidase was blocked using 3%
hydrogen peroxide. After blocking for 5 minutes with
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EROVIC ET AL.
casein, sections were incubated at room temperature with
the appropriate primary antibodies in accord with the conditions described in the following text. Incubation was followed by the application of the Biogenex Super Sensitive
Kit (cat #HK518-YAK; Biogenex, San Ramon, CA) super
enhancer and then the Polymer HRP reagent. After washing in phosphate-buffered saline (PBS), color development
was performed with freshly prepared NovaRed solution
(cat #SK-4800; Vector Laboratories, Burlingame, CA).
Finally, the sections were counterstained with Mayer’s hematoxylin, dehydrated in alcohols, cleared in xylene, and
mounted in Permount (cat #SP15-500; Fisher Scientific,
Fair Lawn, NJ).
Cyclooxygenase-1 and -2 staining
Staining of cyclooxygenase (Cox)-1 and Cox-2 was
performed as follows: 4 lm formalin-fixed paraffin-embedded sections were dewaxed in 5 changes of xylene
and brought down to water through graded alcohols. Endogenous peroxidase and biotin activities were blocked
respectively using 3% hydrogen peroxide and an avidin/
biotin blocking kit (cat # TA-015-BB; Lab Vision, Fremont, CA). After blocking for 15 minutes with 10% normal serum from the species from which the secondary
antibody was obtained, sections were incubated at room
temperature with the appropriate primary antibodies using
the previously optimized conditions (see Table 1). This
was followed with a biotinylated secondary (Vector Laboratories) for 30 minutes and horseradish peroxidase–conjugated ultrastreptavidin labeling reagent (ID Laboratories, London, ON, Canada) for 30 minutes. After
washing in PBS, color development was performed as
described earlier (Table 1).
RESULTS
Twenty patients with sebaceous carcinomas of the head
and neck were eligible for inclusion in the study. Seventeen patients had a carcinoma of the periorbital region, and
3 patients had a sebaceous carcinoma of other head and
neck sites (Table 2). For patients with periorbital sebaceous
carcinomas, the upper eyelid was involved in 7 patients
(35%) and the lower eyelid was involved in 2 patients
(10%). Both eyelids were involved in 2 patients (10%).
The remaining periorbital subsites were the medial canthal
region (n ¼ 3, 15%) and the entire orbit (n ¼ 3, 15%).
The nasal skin (n ¼ 1, 5%) and the scalp (n ¼ 2, 5%)
were the subsites for extraorbital sebaceous carcinomas.
Of the entire cohort (n ¼ 20), 10 were women and 10
were men. The median age at presentation for patients
with periorbital and extraorbital tumors was 72 and 68
years (range, 42–86 years), respectively.
TNM classification
The distribution of tumors by pathologic T and N classifications is as follows: for the periorbital site, 5% (n ¼
1) of the carcinomas were classified as T1 and 80% (n ¼
16) were classified as T3 or T4. At the time of diagnosis,
the tumors presenting at extraorbital sites were classified
as T1 or T2 in 2 cases and 1 patient presented with T4
disease. Of the entire patient cohort, only 2 patients (scalp
HEAD & NECK—DOI 10.1002/HED
Characteristics
No. (%)
Patients
Site
Periorbital
Extraorbital
Sex
Female
Male
Age
Mean, y
Median, y
Range, y
Margins
Positive
Negative
PNI
Postive
Negative
NA
LVI
Positive
Negative
NA
ECE
Positive
NA
20
10 (50)
10 (50)
10 (50)
10 (50)
68
72
42-86
6 (30)
14 (70)
4 (20)
3 (15)
13 (65)
2 (10)
1 (5)
17 (85)
1 (5)
19 (95)
Abbreviations: PNI, perineural invasion; LVI, lymphovascular invasion; ECE, extracapsular
extension.
lesion, 1; both eyelids, 1) had nodal metastases at initial
presentation and distant metastases were not noted at presentation in any of the patients.
Primary treatment
Patient data
644
TABLE 2. Demographic and histopathologic characteristics of
20 patients with sebaceous gland carcinoma.
MAY 2013
Of the 20 patients, 14 were treated with primary surgery; 5 of these 14 patients received postoperative radiotherapy. Six of the 20 patients were treated with primary
radiotherapy. An elective neck dissection was performed
in 1 patient with sebaceous gland carcinoma localized on
the scalp.
Pathology
Positive margins were noted in 6 patients (30%) and
negative margins in 14 patients (70%). Four specimens
(20%) showed perineural invasion, 1 specimen (10%)
showed lymphovascular invasion, and 1 specimen (5%)
showed extracapsular lymph node extension.
Outcomes
The mean and median follow-up times for the entire
cohort were 47 and 28 months, respectively, with a range
of 4 to 145 months. Overall, 11 patients (55%) developed
recurrent disease; of these, 5 patients (45%) developed
local and 6 patients (55%) developed regional recurrence.
At the last follow-up, 7 patients (35%) were alive without
disease, and 3 patients (15%) were alive with disease;
10 patients (50%) died, of which 6 patients died due to
sebaceous gland carcinoma. Of the 11 patients with
recurrence, 6 patients underwent salvage treatment. Four
TARGET
PROTEINS IN SEBACEOUS GLAND CARCINOMAS
cally, 30 samples (88%) and 2 samples (6%) of 34
showed strong and moderate expression for VEGFR-2,
respectively.
Platelet-derived growth factor receptor-a and -b
The majority of the probes were highly positive for
platelet-derived growth factor receptor alpha (PDGFR-a;
n ¼ 27, 78%), with only 1 sample (3%) demonstrating
weak staining and 3 samples (9%) showing moderate
staining. Overexpression of PDGFR-b was found to be
high in 27 probes (78%) and weak or moderate in 2
probes (6%).
Epithelial growth factor receptor
Only 1 sample (3%) was negative for epidermal growth
factor receptor (EGFR), with 26, 5, and 2 samples (76%,
15%, and 6%) showing a high, moderate, and weak
expression, respectively.
Inflammation markers
All specimens were positive for cyclooxygenase-1
(Cox-1) and cyclooxgenase-2 (Cox-2). Cytoplasmatic
expression of Cox-1 was categorized as moderate in 1
specimen (3%) and high in 31 specimens (97%), respectively. Cox-2 expression was weak, moderate, and high in
2, 4, and 28 of all tumor samples (6%, 12%, and 82%),
respectively.
Apoptosis markers
FIGURE 1. The immunostaining of VEGFR2, PDGFRa and -b, EGFR,
Cox-1, Cox-2, Mcl-1, Gli-1, 2-3, Shh, MMP-1, and Gstp in a tissue
microarray of sebaceous gland carcinoma are shown. All
photomicrographs were taken at original magnification, 200. [Color
figure can be viewed in the online issue, which is available at
wileyonlinelibrary.com.]
patients with locally recurrent periorbital sebaceous gland
carcinomas underwent salvage treatment with surgery and
postoperative radiotherapy. Six patients developed regional recurrent disease and were treated either by salvage surgery alone (n ¼ 3) or surgery and radiotherapy
(n ¼ 3). One patient refused further therapy. The median
(mean) survival after treatment for recurrence was 19
(36) months for the entire cohort.
Thirty-three, 15, 17, and 17 samples (100%, 47%,
53%, and 53%) were found to be positive for Mcl-1,
Bclx, Bcl-2a, and p53, respectively. The staining was
localized to the cell cytoplasm. Looking at each of these
proteins independently, Mcl-1 and p53 were highly
expressed in 25 samples (76%) and 15 samples (45%),
whereas only 4 samples (12%) each were highly positive
for Bclx and Bcl-2a. Mcl-1, Bclx, and Bcl-2a were moderately expressed in 4, 2, and 3 samples (12%, 6%, and
9%).
WNT pathway
Expressions of b-catenin, Wisp-1, and Wisp-2 were
localized to the tumor cell cytoplasm, and positive staining was observed in 100% (b-catenin), 30 (91%) (Wisp1), and 11 (34%) (Wisp-2) of the specimens.
Immunohistochemistry
Shh pathway
Original immunohistochemical stains are presented in
Figure 1 and the schematic presentation of negative,
weak, moderate, and high protein expression in sebaceous
gland carcinoma specimens is shown in Figure 2.
All sonic hedgehog (Shh) pathway proteins were
expressed in the tumor cell cytoplasm. Gli-1, Gli-2, and
Gli-3 were detectable in all specimens. Intensity for these
3 proteins was rated as high in 32 (Gli-1), 32 (Gli-2), and
28 (Gli-3) of all investigated samples (97%, 100%, 88%).
Moderate intensity was observed in only 3 specimens (1
for Gli-1 and 2 for Gli-3). Shh was highly overexpressed
in 28 samples (88%), whereas Smo was not positive in
26 samples (76%). Expression of Shh was weak in 2
specimens (6%) and moderate in 1 specimen (3%).
Angiogenesis markers
Vascular endothelial growth factor receptor-2
All investigated samples were positive for vascular endothelial growth factor receptor-2 (VEGFR-2). Specifi-
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EROVIC ET AL.
FIGURE 2. Schematic representation of negative, weak, moderate, intermediate, and strong staining intensity of 33 different proteins in
sebaceous gland carcinoma samples. P/m, primary/metastatic lesion: primary, 1; mets, 2; n, sample number. [Color figure can be viewed in the
online issue, which is available at wileyonlinelibrary.com.]
Cell adhesion markers
Matrix metalloproteinase 1 (MMP-1) was highly overexpressed in all the samples, whereas Keratin-7 was detectable only in 16 samples (50%). For Keratin-7, the intensity was weak or moderate in each of 2 probes (6%),
respectively. CD9 was expressed in the cell membrane of
30 specimens (94%) and its intensity was classified as
moderate in 4 specimens (12%). No expression for CD9
could be found in 2 specimens (6%).
Cell cycle markers
Except for the proteins Cdc2p34 and 14-3-3r, which
were observed in the cell cytoplasm, all remaining proteins belonging to the cell cycle group were found in the
cell nuclei. Cdc2p34 was highly overexpressed in 25 samples (78%) and moderately and weakly expressed in 6
and 1 samples (19% and 3%), respectively. Rb was negative in 18 of all specimens (51%) but was highly overexpressed in 12 samples (34%). Cyclin D1 was positive in
only 10% of samples. Cyclin D1 was classified predominantly as negative (61%) and only 2, 1, and 10 samples
(6%, 3%, and 30%) showed weak, moderate, and high
expression, respectively. The expression of 14-3-3r was
significantly overexpressed in 29 samples (85%), moder646
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ate in 4 samples (12%), and weak in only 1 sample (3%).
The proteins p27 and p21 were predominantly highly positive. In particular, in 27 and 31 of samples (82%, 91%),
high overexpression of p27 and p21 was observed with a
moderate expression in 5 and 2 samples (15%, 6%),
respectively. Bmi-1 was highly overexpressed in all
samples.
AKT-pathway markers
Highly positive staining for pAKT could be observed in
17 samples (52%) and moderate and weak expression was
observed in 9 and 3 of the specimens (27%, 9%), respectively. In contrast, FoxO-1 (Figure 1I) was predominantly
highly overexpressed in 28 of all samples (82%). Its intensity was weak in 1 sample (3%) and moderate in 2
samples (6%). High and weak expression of mTOR
occurred in 23 and 5 specimens (68%, 15%),
respectively.
Cell-specific markers
Glutathione S-transferase pi (Gstp; Figure 1J) was
highly positive in 33 of the 34 samples (79%) and only 1
sample was negative for Gstp.
TARGET
Protein expression in primary and metastatic lesions
We compared the protein expression of all 33 antibodies but we were not able to determine a specific expression pattern related to primary or metastatic lesions in
this cohort.
DISCUSSION
Despite significant improvements in our understanding
of the molecular biology of cancer and its metastatic
behavior, locoregional and distant failures in patients with
sebaceous gland carcinoma are still common.2 The cornerstone of treatment is surgical resection of the primary disease with or without adjuvant radiotherapy.2 Unfortunately,
to date there is no evidence-based data on the management
of patients with distant metastatic disease. In the current
English-language literature there are only 3 case reports
published.3–5 The choice of chemotherapy in these reports
was generally based on a combination of drugs commonly
used in the treatment of head and neck carcinomas.3 The
aim of this study was to determine the antigen profiles of
sebaceous gland carcinomas and to determine if there were
any significant characteristics of those tumors that demonstrated metastatic behavior.
The most impressive immunostaining in sebaceous
gland carcinoma was the expression of VEGFR2, EGFRR2, PDGFR-a, and PDGFR-b. Currently, there are several studies demonstrating that tyrosine kinase receptor
inhibitors are effective in patients with squamous cell carcinomas of the head and neck,6 lung,7 and colon.8 Our
results may provide the rationale for the use of future targeted therapies, such as sunitinib, cetuximab, bevacizumab, and pazopanib against those proteins. These biologic agents have been shown to be effective in the
treatment of head and neck, kidney, lung, and breast
malignancies.6,9–11
In addition, we observed a high rate of expression of
both Cox-1 and Cox-2, particularly Cox-1, in a significant
number of tumor specimens. Multiple studies have been
published that demonstrate an antitumor effect of nonsteroidal anti-inflammatories.12–14 Further research is needed
to determine whether the administration of nonsteroidal
anti-inflammatory drugs (NSAIDs) may provide a benefit
in patients with metastatic sebaceous gland carcinoma.
The hedgehog-signaling cascade is one of the main
pathways in human embryogenesis and organogenesis.
Mutations that inappropriately reactivate this normally
quiescent pathway have been linked to the development
of skin, colon, and lung carcinoma.15,16 In our study, we
observed high expression of the proteins Gli-1, Gli-2,
Gli-3, and Shh. Manipulation of the hedgehog pathway
by selective agonists and antagonists, such as cyclophosphamide, may be effective therapeutic modalities in those
tumors demonstrating overexpression of hedgehog-signaling pathway proteins.16 Research in prostate cancer and
lymphoma has shown that blocking Gli-2 expression with
antisense oligonucleotides in vitro leads to a significant
delay in tumor progression and an increase in paclitaxel
chemosensitivity.17,18
The proteins CD9, MMP-1, and Gstp were also significantly overexpressed in the majority of our sebaceous
gland carcinoma samples. CD9 and MMP-1 are proteins
PROTEINS IN SEBACEOUS GLAND CARCINOMAS
involved in cell motility, invasion, and metastasis.19–21
Selective targeting of CD9 and MMP-1 with specific antibodies has been demonstrated to trigger apoptosis in chemoresistant lung carcinoma cells20 and can suppress the
invasion of tumor cells in a 3-dimensional collagen gel.21
Gstp plays a role in the detoxification of platinum-based
cytotoxic agents and can promote resistance to platinums.
A new target agent named telcyta (TLK)-286 may hold
promise by increasing tumor sensitivity to platinums.
TLK-286 has demonstrated cytotoxic activity on its
own,22 but through its inhibition of Gstp it can enhance
the activity of cisplatin in cancer cells.23 A Gstp inhibitor
such as TLK-286 may hold promise in patients with sebaceous gland carcinoma through its ability to sensitize tumor cells to cisplatin.
Forkhead-box-O (FoxO) transcription factors are
involved in multiple signaling pathways and play critical
roles in a number of physiological processes as well as
tumor pathogenesis.24 FoxO acts as a tumor suppressor,
with loss of FoxO function leading to increased cell survival and a predisposition to cancer development.25 Based
on current knowledge and studies from the past decade,
the development of novel agents, which specifically activate FoxO members, could be useful in the prevention
and treatment of epithelial malignancies in particular.25
Our demonstration of significant FoxO-1 overexpression
in sebaceous gland cancer cells has suggested that development of FoxO-1 target agents may represent a new
effective therapeutic approach.
Another very interesting finding of this study was that
there is no specific protein expression pattern between
primary and metastatic lesions in patients with sebaceous
gland carcinoma. Initially we hoped to find a target protein, particularly for metastatic lesions. Conversely, no
differential protein expression means that targeting the
primary tumor may simultaneously target distant metastases in patients with advanced-stage disease.
Although it is recognized that these results are preliminary and that further in vitro and in vivo data are
required, we have identified several potentially important
protein targets and a target pathway that may be therapeutically exploited for the treatment of sebaceous gland
carcinoma. The use of agents that target these proteins
and pathways may provide a much-needed systemic treatment for patients with metastatic sebaceous gland
carcinoma.
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