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May 1982]
Copper- and Zinc-containing
Superoxide Dismutase, Manganese-containing
Superoxide Dismutase, Catalase, and Glutathione Peroxidase in Normal
and Neoplastic Human Cell Lines and Normal Human Tissues1
Stefan L. Marklund, N. Gunnar Westman, Erik Lundgren, and Goran Roos
Departments of Clinical Chemistry [S. L. M.], Oncology [N. G. WJ, and Pathology [E. L., G. R.J, University Hospital of Urnea, S-901 85 Umeà , Sweden
ABSTRACT
Copper- and zinc-containing Superoxide dismutase, man
ganese-containing Superoxide dismutase, catalase, and glutathione peroxidase form the primary enzymic defense against
toxic oxygen reduction metabolites. Such metabolites have
been implicated in the damage brought about by ionizing
radiation, as well as in the effects of several cytostatic com
pounds.
These enzymes were analyzed in 31 different human normal
diploid and neoplastic cell lines and for comparison in 15
normal human tissues. The copper- and zinc-containing superoxide dismutase appeared to be slightly lower in malignant cell
lines in general as compared to normal tissues. The content of
manganese Superoxide dismutase was more variable than the
content of the copper- and zinc-containing enzyme. Contrary
to what has been suggested before, this enzyme did not appear
to be generally lower in malignant cells compared to normal
cells. One cell line, of mesothelioma origin (P27), was ex
tremely abundant in manganese-containing
Superoxide dis
mutase; the concentration was almost an order of magnitude
larger than in the richest normal tissue.
Catalase was very variable both among the normal tissues
and among the malignant cells, whereas glutathione peroxi
dase was more evenly distributed. In neither case was a general
difference between normal cells and tissues and malignant
cells apparent.
The myocardial damage brought about by doxorubicin has
been linked to toxic oxygen metabolites; particularly, an effect
on the glutathione system has been noted. The heart is one of
the tissues which have a low concentration of enzymes which
protect against hydroperoxides. However, the deviation from
other tissues is probably not large enough to provide a full
explanation for the high doxorubicin susceptibility.
In the present survey, no obvious relationship between gen
erally assumed resistance to ionizing radiation or to radicalproducing drugs and cellular content of any of the enzymes
could be demonstrated.
INTRODUCTION
Enzymes protecting against peroxides, and against the superoxide radical in particular, have recently received much
attention in connection with malignant tumors. There are 2
main reasons for this, (a) Much of the damaging effects of
ionizing radiation are due to reactive free radicals, and the
effects are enhanced in the presence of oxygen. Under several
1 This study was supported by the Swedish Medical Research Council, Grant
04761 and the Lions Research Foundation, Department of Oncology, University
of Umeâ,Umea, Sweden.
Received November 6, 1981 ; accepted February 3, 1982.
MAY
1982
experimental conditions, enzymes scavenging the Superoxide
radical, the Superoxide dismutases, appear to protect against
ionizing radiation (e.g., Refs. 26, 28, 40, 42, 47, 49, 60, and
63). (b) Because several of the commonly used cytostatic
agents, e.g., the anthracyclines (2, 17, 22, 30, 43) and bleomycin (29, 53), appear to exert their effects by way of reactive
free radicals. Again, Superoxide dismutase has been protective
in some instances (29, 30, 53).
Much has been written about the Superoxide dismutases in
malignant cells and tumors during the last few years. However,
the conclusions drawn have in general been based on few
experimental observations, and those mainly on animal tumors
(4, 45, 46, 48, 52). The purpose of the present investigation
was to make a more comprehensive study of the Superoxide
dismutases in normal and neoplastic human cell lines. For
comparison, we analyzed the enzymes in normal human tis
sues.
We also analyzed two enzymes that scavenge hydroperox
ides, catalase and glutathione peroxidase. Much of the toxicity
of the Superoxide radical is believed to go by way of the
hydroxyl radical formed in a transition metal ion-catalyzed
reaction with hydrogen peroxide (20, 27, 38, 60). The com
bined efforts of the Superoxide dismutases and hydrogen per
oxide-scavenging enzymes therefore appear to be of impor
tance for a reduction of the free radical toxicity within cells.
MATERIALS
AND METHODS
Unless otherwise specified, the analyses were performed
at 25°.
Tissues and Tumors. Human tissues from accident or suicide victims
without known physical diseases were obtained within 24 hr after death
from the Department of Forensic Medicine. The tissues were kept at
—
80° before assay. They were homogenized in an Ultra-Turrax with
20 volumes of 10 mM potassium phosphate (pH 8.0) plus 30 nriM KCI.
The homogenates were then sonicated with a Branson B30 under
cooling with ice. After extraction for 30 min at 4°, the homogenates
were centrifuged (20,000 x g, 15 min), and enzyme and protein
analysis was performed on the supernatants.
Cell Lines. The material consisted of 31 human cell lines of different
origins. Most lines are well known and are characterized in a number
of earlier investigations. The lines and their respective derivation are
listed in Table 2 together with selected references. The mesothelioma
cell lines were established from pleural effusions and have neoplastic
properties as judged by chromosome analysis and cell growth char
acteristics. These lines will soon be reported on separately. The JC-1
line, derived from a giant cell tumor, has reached passage 21 and
cannot yet be considered as a permanent cell line. The line has a
mesenchymal cell-like appearance and is aneuploid.
The cells were grown in Eagle's minimal essential medium (anchor
age-dependent
lines) or Ham's F-10 medium (suspension-growing
lines) with 10% fetal calf serum, supplemented with benzyl penicillin
and streptomycin in air containing 5% CO? and saturated with M..O
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S. L. Marklund et al.
The cells were harvested 3 to 7 days after explantation when they were
in a near-confluent state. They were washed twice with 0.15 M NaCI,
and the monolayer cells were then scraped off with a bent-glass pipet
in 0.15 M NaCI with 3 ÕTIM
diethylenetriaminepentaacetic
acid. The
suspensions were centrifugea, and the cells were kept as pellets at
—
80° until assay. For assay, the cells were sonicated under cooling
with ice in 10 mM potassium phosphate (pH 7.4) containing 30 mM
KCI. After extraction for 30 min, the homogenates were centrifuged
(20,000 x g, 15 min), and enzyme and protein analysis was performed
on the supernatants.
Specific Assay of Hemoglobin in Tissue Homogenates and Com
pensation for Erythrocyte Enzymes. Erythrocytes contain large
amounts of 3 of the enzymes of the present investigation. The content
of glutathione peroxidase and especially of catalase was larger than
that of several of the tissues. It was therefore necessary to compensate
for enzyme activity in the homogenates contributed by erythrocytes.
The hemoglobin was specifically determined in terms of the differential
protective activity of serum (haploglobin) and egg white on the peroxidatic activity
contamination
of the hemoprotein (35). The corrections for blood
in tissue homogenates were made with the actual en-
zymic activities of erythrocytes from the individual persons.
Superoxide Dismutase Analysis. Superoxide dismutase was deter
mined in terms of its ability to catalyze the disproportionation of O2~ in
alkaline aqueous solution. The disproportionation
was directly studied
in a spectrophotometer,
essentially as described before (34), the
difference being that both CuZn Superoxide dismutase2 and Mn superoxide dismutase were assayed at pH 9.50 and that
used for the distinction between the enzymes. One
defined as the activity that brings about a decay in
at a rate of 0.1 s"1 in 3 ml buffer. It corresponds to
3 mM cyanide was
unit in the assay is
Cv concentration
8.3 ng human and
to 4.1 ng bovine CuZn Superoxide dismutase and 65 ng bovine Mn
Superoxide dismutase. The pure human manganese-containing enzyme
has not been investigated with this assay, but its specific activity is
probably similar to that of the bovine enzyme. The xanthine oxidasecytochrome c assay for Superoxide dismutase works at physiological
conditions, i.e., neutral pH and low O2~ concentration (39). When
bovine and human enzymes are analyzed, 1 unit in the present assay
corresponds to 0.024 unit CuZn Superoxide dismutase and 0.24 unit
Mn Superoxide dismutase, respectively, in the "xanthine oxidase"
assay. The present assay is thus about 10 times more sensitive for
CuZn Superoxide dismutase activity than for Mn Superoxide dismutase
activity. Staining for Superoxide dismutase in agarose gel plates was
performed with a slight modification of the method of Beauchamp and
Fridovich (3).
Catalase. For catalase assay, 1% ethanol was added to the homog
enates to prevent catalase Compound II formation. The activity was
analysed with a Clark oxygen electrode essentially as described by Del
Rio ef al. (9). Homogenates were added to 3 ml deaerated 10 mM
potassium phosphate plus 0.1 diethylenetriaminepentaacetic
acid (pH
7.4) containing 15 mw H2O2. Catalase activity was calculated from the
initial rate of O2 liberation. One unit is defined as the amount that
disproportionates
1% of the hydrogen peroxide in 1 min. The method
is about 50 times more sensitive than are conventional spectrophotometric or titration methods.
Glutathione Peroxidase. Glutathione peroxidase was assayed by
the method of Gunzler et al. (19) with some modifications. Homoge
nates were added to a final volume of 500 jil 50 mM potassium
phosphate plus 2 mM diethylenetriaminepentaacetic
acid (pH 7.0) with
0.16 mM fe/t-butyl hydroperoxide at 37°.ferf-Butyl hydroperoxide was
used instead of
hence a higher
determined, the
cyanide and 8.7
HjO2 because much lower blanks were obtained and
sensitivity. When the activity of erythrocytes was
hemolysates were treated with 1 mw potassium ferrimM NaCn to inhibit the activity of hemoglobin. Hemo-
2 The abbreviations used are: CuZn Superoxide dismutase, copper- and zinccontaining Superoxide dismutase; Mn Superoxide dismutase, managanese-containing Superoxide dismutase.
1956
lysates were also analyzed without these additions, and the results
were used for the compensations for blood contamination in tissue
homogenates.
Protein Analysis. For protein analysis, Coomassie Brilliant Blue G250 was used (6). Human serum albumin was used for standardization.
This sensitive convenient method was compared with the more estab
lished technique of Lowry ef al. (31). Human tissue homogenates
(pancreas, lymphatic node, muscle, lung, heart, renal cortex, renal
medulla, thyroid gland, liver, brain white matter, brain gray matter,
adipose tissue, and spleen) were analyzed with both methods stand
ardized with human serum albumin. The results were very similar, the
ratio between the Lowry results and the results of the present method
being 1.12 ±0.14 (S.D.) (range, 0.98 to 1.42).
RESULTS AND DISCUSSION
The results of enzyme analysis in tissues (Table 1) are
presented both as units/mg protein and units/g, wet weight.
The latter mode should better describe the protective capacity
of the enzymes within the cells since it is roughly proportional
to the concentration of the enzymes. The units/mg protein
results are given to enable comparison with the cell lines where
a representative "wet weight" is difficult to obtain.
Superoxide Dismutase. Superoxide dismutase catalyzes the
disproportionation
of the Superoxide anión radical, 2C-2~ +
2H+ -»O2 + H2Oj (39). The enzyme is found in all aerobic
cells and is believed to exert an important protective function
against the toxicity of oxygen (14). In eukaryotic cells, 2 forms
of the enzyme are generally found, one cytosolic and mitochondrial enzyme containing copper and zinc and one mito
chondria! enzyme containing manganese (62). In primates, the
Mn Superoxide dismutase is possibly also found in the cyto
plasm (37). The enzymes should be determined separately
because of their partly different localizations; their role in
protection of various targets in the cells against various sources
of Superoxide radical may differ.
CuZn Superoxide dismutase activity was found in all investi
gated tissues (Table 1) and cell lines (Table 2). There are
comparatively small differences in this activity within the
groups. Although only in a few cases can a direct comparison
be made between a cell line and tissue of origin, it appeared
that the content of CuZn Superoxide dismutase is on the whole
lower in the cell lines than in the normal tissues. This has been
reported before (15, 45, 46, 48, 52, 57). The enzyme activity
in the renal cancer cell line HCV ¡slower than in the kidneys.
On the other hand, the activities of the various lymphoma cells
were not lower than that of the lymphocytes and the lymphatic
node but were in a few cases higher. A similar observation has
been reported before (66). Among the cultivated cells, lympho
cytes and polymorphonuclear cells (Table 2), no general dif
ference was seen in CuZn Superoxide dismutase activity be
tween normal diploid cells and neoplastic cells.
There seems to be a relationship between a high metabolic
activity in tissues, and hence many mitochondria, and much
Mn Superoxide dismutase; liver, kidney, heart, adrenal gland,
and brain gray matter possess much enzyme (Table 1). There
are much larger differences in Mn Superoxide dismutase activ
ity than in CuZn Superoxide dismutase activity among cultivated
cells (Table 2). The larger differences for this enzyme may be
related to its reported inducibility (7, 11, 54, 65). Several
investigators have found no (45, 52) or very little (4, 46, 48,
59) Mn Superoxide dismutase in various tumors and neoplastic
CANCER
RESEARCH
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VOL. 42
Enzyme Defenses in Normal and Neoplastia Cells and Tissues
Table 1
CuZn Superoxide dismutase, Mn Superoxide dismutase, catatase, and glutathione peroxidase in normal human tissues
Tissues from 2 individuals, A and B, were analyzed. The figures are corrected for enzymes contributed by blood contamination.
CuZn Superoxide dismutase
Mn Superoxide dismu
tase
Catalase
Glutathione peroxidase
/mg pro
/mg pro
/mg pro
wt1wet
protein1901201919140871209073502001604311053
tein31350a026404530282.27.41811305.11126673.77.20.91.216248.9139.111U/g,
tein1,3001,5009901,30043011030070022056"012010012021018054"036250011Ö3b200270
wt83,000114,000337,000431
wet
tein1,2001,5006761350470850420310952002501602601701603403302802007991540480730570200220U/g,
wt79,0001
wet
wt2,1002,900001.6003,2002,0001,1005902605106406501,0002605801,0002,70014038011014035066013022054
wet
LiverABErythrocytesABRenal
,15008,9006.5006,5006,3006.40
20,00023,00021
,00019,3008,20012.30015,1008,2007.800604.0003,4006.3008,7006,6001
,00022,00037,00038,0001
cortexABAdrenal
glandBRenal
medullaABSpleenABLymphatic
6,00013,0001
,00014,0009,2009,8009,6008,7008.60013,00013,0001
1
nodeAPancreasABLungABHeartABSkeletal
,800"01.3001,300002006100"30001.1001,800U/mg
muscleABThyroid
,0001
1
,0009,50010,0001
1
glandABBrain
matterABBrain
gray
2,0001
3,0001
matterABAdipose
white
,10009,6001,200800U
tissueABU
'' 0, corrections about as large as the total catalase activity.
6 Corrections were large, 75 to 90% of the total activity, making the results less reliable.
cell lines. The idea has been advanced that loss of Mn superoxide dismutase is intimately related to the cancerous phenotypes (46). This proposition was based on analysis of relatively
few cell lines and tumors, and most investigations were made
on animal material. Our results on human cell lines (Table 2)
do not support the proposition. We find highly variable amounts
of Mn Superoxide dismutase in the cultivated neoplastic cells
and in some cases very large amounts. A similar finding in
human solid tumors was recently reported (64). The difference
MAY 1982
may be due to the fact that Mn Superoxide dismutase is
apparently found in both cytosol and mitochondrial matrix in
primates (37) whereas in other species the enzyme is found
only in mitochondrial matrix. Comparison with tissue of origin
can be made only in a few cases. The cell line HCV derived
from a renal adenocarcinoma possessed about as much en
zyme as did the kidney. The various lymphatic neoplastic cells
varied in enzyme content but possessed about the same quan
tities of enzymes as did the normal lymphocytes and a little less
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S. L. Marklund et al.
Table 2
CuZn Superoxide dismutase, Mn Superoxide dismutase, catatase, and glutathione peroxidase in human
normal diploid and human neoplastic cell lines
A few of the cell lines have not been described in publications.
Roos. and Lundgren, and zur Hausen.
lineAnchorage
They were taken into culture by Fogh.
superone per
oxide dismu
Superoxide
oxidase
tase (U/mg
dismutase (U
(U/mg
(U/mg
protein)12148575472.05.3324.1241264023013Catalase
mi i
protein)1301301108318013014014016072240250200205Mn
protein)749382735.026352925084185.530NDCGlutathi
protein)4548233776469.2487039627.773NORef.2121231116abt>t>
Cell
dependentFibroblastsSkin(HED21)Lung
20)Adult
(HEL
skinEmbryonal
lungEndothelial
cordECFibroblasts
cells, umbilical
transformed)WÕ38
(SV40
3Ovarian clone Va1
epithelial,aneuploidA7A10Cervical
adenocarcinoma,
adenocarcinomaHeLaRenal
carcinoma, epithelial, aneu
ploidHCV29Malignant
mesotheliomaP7
aneuploid)P31
(epithelial
aneuploid)P27
(epithelial
aneuploid)P30
(fibroblastoid
(fibroblastoid
aneuploid)CuZn
Giant cell tumor (fibroblastoid
ploid)
JC1
240
b
250
40
7.7
6
200
230
30
73
tb5550
205170120120143
131.43.25.74.4NDCNO3528NDNO62
NDND30170NDNDb
aneu
Amnionic fluid (epithelial aneuploid)
Amnion U
Glioma
251 MG
Suspension growing
Normal blood lymphocytes (n •21)
Normal blood polymorphonuclear
kocytes (n = 4)
Lymphoblastoid
LÕA
30"*114
±
.42.1
±1
±12126
leu
±0.34185.5
diploid B-cells
160
22
62
36
Myeloma, plasmablasts
RPMI8226
300
5.1
67
65
Plasma cell leukemia lymphoblastoid
B-cells
IB
150
3.0
79
140
180
3.2
32
1.5
24
Lymphocytic leukemia T-lymphoblast
Jurkat
CCRF-CEM
Molt 4
300
240
170
8.7
3.4
1.3
110
240
110
6.1
5.6
ND
13
41
Histiocytic lymphoma
Platz
U 937310
2508.8
4.724
590122504456
36
Burkitt's lymphoma, B-lymphoblast
aneuploid
Namalwa
1958
CANCER
RESEARCH
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VOL. 42
Enzyme Defenses in Normal and Neoplastia Cells and Tissues
Table 2—Continued
lineMyeloid
Cell
super-oxide
per-oxidase(U/mgprotein)64470Ref.82532
dismu-tase
Superoxidedismutase
(U/mgprotein)190220210Mn
(U/mg
protein)3.66.83.6Catalase(U/mgprotein)92025043Glutathi-one
leukemiaHL
promyelocytesKG
60
myeloblastsK
1
562-4 blastic crisisCuZn
Cultured by J. Fogh.
6 Cultured by G. Roos and L. Lundgren.
c ND, not determined.
Mean ±SD.
8 Cultured by H. zur Hausen.
than the studied lymphatic node. The promyelocytic leukemia
cell line, HL 60, possessed a little more Mn Superoxide dis
mutase than did mature normal polymorphonuclear
cells. A
comparison of the normal fibroblasts and the SV40-transformed fibroblast, Wi38 clone Va13, shows a decrease in Mn
Superoxide dismutase in the transformed cells. This is in accord
with what has been reported before (65). The mesothelioma
cell lines were exceptional. All possessed much Mn Superoxide
dismutase, the P27 extremely much, almost an order of mag
nitude more than the richest tissue. The method for Superoxide
dismutase analysis is 10 times less sensitive for Mn Superoxide
dismutase than for CuZn Superoxide dismutase, which means
that the Mn Superoxide dismutase figures should be multiplied
by 10 in order to make them comparable with the CuZn
Superoxide dismutase figures. These cells are thus extremely
well equipped with Superoxide dismutases. They should form
an interesting object for the study of the protective importance
of the Superoxide dismutases under various conditions. We
have thus far not been able to obtain normal mesothelial cells
for comparison. The mesothelioma cell lines and several of the
tissues were also studied by electrophoresis and subsequent
staining for Superoxide dismutase. This was done in order to
check the unexpectedly large amounts of Mn Superoxide dis
mutases found. In every case, the semiquantitative evaluation
of the stained gel agreed with the presented enzymic activities.
The electrophoretic mobilities of the mesothelioma CuZn superoxide dismutase and Mn Superoxide dismutase appeared
equal to those of the normal tissues.
The content of CuZn and Mn Superoxide dismutase in the
tissues shows no correlation to generally assumed radiosensitivities or to sensitivity to cytostatic drugs. For example,
radiosensitive tissues like lung and lymphatic node contain
about the same amount of Superoxide dismutase (per g, wet
weight) as do less sensitive tissues like heart, skeletal muscle,
and brain gray matter. This conclusion is also supported by the
findings among the cell lines. The activities of the generally
radiosensitive lymphocytic cell lines and normal blood lympho
cytes are not much different from those of nonlymphocytic
cells. The findings of very large amounts of Mn Superoxide
dismutase in the mesothelioma cell lines are difficult to evaluate
with respect to clinical radioresponsiveness,
since although
these tumors are considered less responsive the clinical ex
perience of treatment is limited.
Catalase. Catalase catalyzes the disproportionate
of hy
drogen peroxide, 2H2O2 -» O2 + H2O, which is formed by
ionizing radiation and probably by the potentially radical-pro
ducing cytostatic drugs. Most tissues (Table 1) possess very
MAY 1982
little catalase, which makes analysis with conventional methods
difficult. This problem was overcome by the present very sen
sitive method. Another problem is the very high catalase con
tent of erythrocytes which makes correction for blood contam
ination necessary. This was done for the tissues of the present
investigation. In a few blood-rich catalase-low tissues, the
corrections were so large that there was no significant remain
ing activity. Very large differences are found in catalase activity
among the tissues and cell lines. Most neoplastic cell lines
were low in catalase activity although some possessed large
amounts, like the promyelocytic leukemia cell line HL 60, the
histiocytic lymphoma cell line U 937, the epithelial HeLa cell
line, and the acute T-lymphocytic leukemia lines Jurkat, CCRFCEM, and Molt 4.
The erythroleukemia cell line derived from a patient with
chronic myeloid leukemia in blastic crisis, K 562-4, was poor
in catalase in contrast to the very high activity found in normal
erythrocytes. No general difference between nontransformed
and neoplastic cells and tissues can be seen. Several authors
have reported very low catalase activities in tumor cells (5, 61 ),
although ample amounts have also been found (51 ). A corre
lation between catalase activity and radiation resistance has
been claimed (33, 61), but the general view appears to be that
this activity is of minor importance (58). Neither is such a
correlation apparent in the present material. For example, the
lymphatic tissues spleen and lymphatic node contain even
more catalase than do "radioresistant"
tissues like skeletal
muscle and brain gray matter, and the liver contains very much
catalase (as well as the other enzymes) although it is not the
most radioresistant tissue. Among the cell lines, the lympho
cytic cells do not differ from nonlymphocytic cells. An interest
ing finding is, however, the difference in catalase and in glutathione peroxidase activity of the 2 histiocytic cell lines Platz
and U 937. It is a well-known clinical experience that some
histiocytic lymphomas may be highly radiosensitive while oth
ers respond more sluggishly. It is also known that different
subpopulations of lymphocytes have different radiosensitivities,
the T-helper lymphocytes being much more radioresistant than
are B-lymphocytes and T-suppressor cells (12). Our T-lymphoblast cell lines also have variable amounts of catalase.
Glutathione Peroxidase. Glutathione peroxidase removes
hydrogen peroxide by catalyzing its reaction with glutathione
(GSH) (H2O2 + 2GSH -»2H2O2 + GSSG). In addition to H2O2,
the enzyme catalyzes the reduction of other hydroperoxides
and thus has a broader protective spectrum than does catalase.
Compared with catalase, there were smaller differences in
glutathione peroxidase activity between the various tissues
1959
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S. L. Marklund et al.
(Table 1) and cell lines (Table 2). The general impression is
that the activities of the neoplastia cell lines are evenly distrib
uted within the range of the normal tissues. The results are in
accordance with previous reports of significant amounts of
glutathione peroxidase in neoplastic cells (5, 48). As for the
other enzymes, there is no apparent relation between this
activity and generally assumed resistance towards radiation
and potentially radical-producing cytostatics.
Doxorubicin Toxicity and Enzyme Activity. The cardiotoxic
action of doxorubicin has been linked to toxic oxygen metab
olites. An increased lipid peroxidation (43) and a decreased
glutathione peroxidase content (10) has been found in the
heart after doxorubicin administration. Animals raised on a
selenium-deficient diet become deficient in the selenium en
zyme glutathione peroxidase. They are much more susceptible
to doxorubicin toxicity (10). Whereas ample amounts of the
Superoxide dismutases are found in the heart (Table 1), the
activities of enzymes protecting against hydroperoxides, glu
tathione peroxidase and catalase, are very low. This fact may
contribute to but not provide a full explanation for the high
doxorubicin susceptibility of the heart since equally low activi
ties are seen in skeletal muscle and in thyroid gland. Brain
tissue is excluded from this comparison since the drug does
not penetrate the blood-brain barrier. Whereas a low catalase
content in mouse heart compared to mouse liver recently was
reported, almost equal glutathione peroxidase activities were
found (10). Another recent investigation of the tissue distribu
tion of catalase and glutathione peroxidase in the mouse (18)
resulted, however, in findings very similar to those presented
here for human tissues.
Determinants of Enzyme Activities. There are considerable
differences in the content of the enzymes in the tissues. This
must to a large extent be due to the demands created by
metabolic specialization within the cells and to various environ
mental factors such as different oxygénation and exposure to
various metabolites. The various cell lines were cultivated un
der comparatively similar environmental conditions created by
the nutritive cell culture medium. However, the differences in
enzyme content in the cell lines appear to be as large as the
differences between various tissues. This indicates that the
inherent character and metabolic specialization of cells are the
main determinants of the enzymic activities and that such
characters are retained in the cultivated cells.
Conclusions. Toxic oxygen reduction metabolites have been
implicated in the damage brought about by ionizing radiation,
especially in the presence of oxygen, and in the effects of
several cytostatic compounds. CuZn Superoxide dismutase,
Mn Superoxide dismutase, catalase, and glutathione peroxi
dase form the primary enzymic defense against toxic oxygen
reduction products. The enzymes might conceivably be of
importance both for the response to ionizing radiation and for
the response to some cytostatics. No general difference in any
of these enzymes between normal cells and tissues and neo
plastic cell lines could be demonstrated in the present investi
gation, nor with certainty could the activities be correlated with
resistance towards ionizing radiation and potentially radicalproducing drugs.
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Copper- and Zinc-containing Superoxide Dismutase,
Manganese-containing Superoxide Dismutase, Catalase, and
Glutathione Peroxidase in Normal and Neoplastic Human Cell
Lines and Normal Human Tissues
Stefan L. Marklund, N. Gunnar Westman, Erik Lundgren, et al.
Cancer Res 1982;42:1955-1961.
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