[CANCER RESEARCH 42, 1955-1961, May 1982]OO08-5472/82/OO42-0000$02.00

Copper- and Zinc-containing Superoxide Dismutase, Manganese-containing

Superoxide Dismutase, Catalase, and Glutathione Peroxidase in Normaland 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, manganese-containing Superoxide dismutase, catalase, and gluta-

thione peroxidase form the primary enzymic defense againsttoxic oxygen reduction metabolites. Such metabolites havebeen implicated in the damage brought about by ionizingradiation, as well as in the effects of several cytostatic compounds.

These enzymes were analyzed in 31 different human normaldiploid and neoplastic cell lines and for comparison in 15normal human tissues. The copper- and zinc-containing super-

oxide dismutase appeared to be slightly lower in malignant celllines in general as compared to normal tissues. The content ofmanganese Superoxide dismutase was more variable than thecontent of the copper- and zinc-containing enzyme. Contrary

to what has been suggested before, this enzyme did not appearto be generally lower in malignant cells compared to normalcells. One cell line, of mesothelioma origin (P27), was extremely abundant in manganese-containing Superoxide dis

mutase; the concentration was almost an order of magnitudelarger than in the richest normal tissue.

Catalase was very variable both among the normal tissuesand among the malignant cells, whereas glutathione peroxidase was more evenly distributed. In neither case was a generaldifference between normal cells and tissues and malignantcells apparent.

The myocardial damage brought about by doxorubicin hasbeen linked to toxic oxygen metabolites; particularly, an effecton the glutathione system has been noted. The heart is one ofthe tissues which have a low concentration of enzymes whichprotect against hydroperoxides. However, the deviation fromother tissues is probably not large enough to provide a fullexplanation for the high doxorubicin susceptibility.

In the present survey, no obvious relationship between generally assumed resistance to ionizing radiation or to radical-producing drugs and cellular content of any of the enzymescould be demonstrated.

INTRODUCTION

Enzymes protecting against peroxides, and against the su-peroxide radical in particular, have recently received muchattention in connection with malignant tumors. There are 2main reasons for this, (a) Much of the damaging effects ofionizing radiation are due to reactive free radicals, and theeffects are enhanced in the presence of oxygen. Under several

1This study was supported by the Swedish Medical Research Council, Grant

04761 and the Lions Research Foundation, Department of Oncology, Universityof Umeâ,Umea, Sweden.

Received November 6, 1981 ; accepted February 3, 1982.

experimental conditions, enzymes scavenging the Superoxideradical, the Superoxide dismutases, appear to protect againstionizing radiation (e.g., Refs. 26, 28, 40, 42, 47, 49, 60, and63). (b) Because several of the commonly used cytostaticagents, e.g., the anthracyclines (2, 17, 22, 30, 43) and bleo-

mycin (29, 53), appear to exert their effects by way of reactivefree radicals. Again, Superoxide dismutase has been protectivein some instances (29, 30, 53).

Much has been written about the Superoxide dismutases inmalignant cells and tumors during the last few years. However,the conclusions drawn have in general been based on fewexperimental observations, and those mainly on animal tumors(4, 45, 46, 48, 52). The purpose of the present investigationwas to make a more comprehensive study of the Superoxidedismutases in normal and neoplastic human cell lines. Forcomparison, we analyzed the enzymes in normal human tissues.

We also analyzed two enzymes that scavenge hydroperoxides, catalase and glutathione peroxidase. Much of the toxicityof the Superoxide radical is believed to go by way of thehydroxyl radical formed in a transition metal ion-catalyzed

reaction with hydrogen peroxide (20, 27, 38, 60). The combined efforts of the Superoxide dismutases and hydrogen peroxide-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 victimswithout known physical diseases were obtained within 24 hr after deathfrom 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 undercooling with ice. After extraction for 30 min at 4°, the homogenates

were centrifuged (20,000 x g, 15 min), and enzyme and proteinanalysis was performed on the supernatants.

Cell Lines. The material consisted of 31 human cell lines of differentorigins. Most lines are well known and are characterized in a numberof earlier investigations. The lines and their respective derivation arelisted in Table 2 together with selected references. The mesotheliomacell lines were established from pleural effusions and have neoplasticproperties as judged by chromosome analysis and cell growth characteristics. These lines will soon be reported on separately. The JC-1

line, derived from a giant cell tumor, has reached passage 21 andcannot yet be considered as a permanent cell line. The line has amesenchymal cell-like appearance and is aneuploid.

The cells were grown in Eagle's minimal essential medium (anchorage-dependent lines) or Ham's F-10 medium (suspension-growing

lines) with 10% fetal calf serum, supplemented with benzyl penicillinand streptomycin in air containing 5% CO? and saturated with M..O

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The cells were harvested 3 to 7 days after explantation when they werein 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 ÕTIMdiethylenetriaminepentaacetic acid. Thesuspensions 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 mMKCI. After extraction for 30 min, the homogenates were centrifuged(20,000 x g, 15 min), and enzyme and protein analysis was performedon the supernatants.

Specific Assay of Hemoglobin in Tissue Homogenates and Compensation for Erythrocyte Enzymes. Erythrocytes contain largeamounts of 3 of the enzymes of the present investigation. The contentof glutathione peroxidase and especially of catalase was larger thanthat of several of the tissues. It was therefore necessary to compensatefor enzyme activity in the homogenates contributed by erythrocytes.The hemoglobin was specifically determined in terms of the differentialprotective activity of serum (haploglobin) and egg white on the perox-

idatic activity of the hemoprotein (35). The corrections for bloodcontamination 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 studiedin a spectrophotometer, essentially as described before (34), thedifference being that both CuZn Superoxide dismutase2 and Mn super-

oxide dismutase were assayed at pH 9.50 and that 3 mM cyanide wasused for the distinction between the enzymes. One unit in the assay isdefined as the activity that brings about a decay in Cv concentrationat a rate of 0.1 s"1 in 3 ml buffer. It corresponds to 8.3 ng human and

to 4.1 ng bovine CuZn Superoxide dismutase and 65 ng bovine MnSuperoxide dismutase. The pure human manganese-containing enzyme

has not been investigated with this assay, but its specific activity isprobably similar to that of the bovine enzyme. The xanthine oxidase-

cytochrome c assay for Superoxide dismutase works at physiologicalconditions, i.e., neutral pH and low O2~ concentration (39). When

bovine and human enzymes are analyzed, 1 unit in the present assaycorresponds to 0.024 unit CuZn Superoxide dismutase and 0.24 unitMn Superoxide dismutase, respectively, in the "xanthine oxidase"

assay. The present assay is thus about 10 times more sensitive forCuZn Superoxide dismutase activity than for Mn Superoxide dismutaseactivity. Staining for Superoxide dismutase in agarose gel plates wasperformed with a slight modification of the method of Beauchamp andFridovich (3).

Catalase. For catalase assay, 1% ethanol was added to the homogenates to prevent catalase Compound II formation. The activity wasanalysed with a Clark oxygen electrode essentially as described by DelRio ef al. (9). Homogenates were added to 3 ml deaerated 10 mMpotassium phosphate plus 0.1 diethylenetriaminepentaacetic acid (pH7.4) containing 15 mw H2O2. Catalase activity was calculated from theinitial rate of O2 liberation. One unit is defined as the amount thatdisproportionates 1% of the hydrogen peroxide in 1 min. The methodis about 50 times more sensitive than are conventional spectrophoto-

metric or titration methods.Glutathione Peroxidase. Glutathione peroxidase was assayed by

the method of Gunzler et al. (19) with some modifications. Homogenates were added to a final volume of 500 jil 50 mM potassiumphosphate plus 2 mM diethylenetriaminepentaacetic acid (pH 7.0) with0.16 mM fe/t-butyl hydroperoxide at 37°.ferf-Butyl hydroperoxide was

used instead of HjO2 because much lower blanks were obtained andhence a higher sensitivity. When the activity of erythrocytes wasdetermined, the hemolysates were treated with 1 mw potassium ferri-cyanide and 8.7 mM NaCn to inhibit the activity of hemoglobin. Hemo-

2 The abbreviations used are: CuZn Superoxide dismutase, copper- and zinc-

containing Superoxide dismutase; Mn Superoxide dismutase, managanese-con-taining Superoxide dismutase.

lysates were also analyzed without these additions, and the resultswere used for the compensations for blood contamination in tissuehomogenates.

Protein Analysis. For protein analysis, Coomassie Brilliant Blue G-250 was used (6). Human serum albumin was used for standardization.This sensitive convenient method was compared with the more established technique of Lowry ef al. (31). Human tissue homogenates(pancreas, lymphatic node, muscle, lung, heart, renal cortex, renalmedulla, thyroid gland, liver, brain white matter, brain gray matter,adipose tissue, and spleen) were analyzed with both methods standardized with human serum albumin. The results were very similar, theratio between the Lowry results and the results of the present methodbeing 1.12 ±0.14 (S.D.) (range, 0.98 to 1.42).

RESULTS AND DISCUSSION

The results of enzyme analysis in tissues (Table 1) arepresented both as units/mg protein and units/g, wet weight.The latter mode should better describe the protective capacityof the enzymes within the cells since it is roughly proportionalto the concentration of the enzymes. The units/mg proteinresults are given to enable comparison with the cell lines wherea representative "wet weight" is difficult to obtain.

Superoxide Dismutase. Superoxide dismutase catalyzes thedisproportionation 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 functionagainst the toxicity of oxygen (14). In eukaryotic cells, 2 formsof the enzyme are generally found, one cytosolic and mito-chondrial enzyme containing copper and zinc and one mitochondria! enzyme containing manganese (62). In primates, theMn Superoxide dismutase is possibly also found in the cytoplasm (37). The enzymes should be determined separatelybecause of their partly different localizations; their role inprotection of various targets in the cells against various sourcesof Superoxide radical may differ.

CuZn Superoxide dismutase activity was found in all investigated tissues (Table 1) and cell lines (Table 2). There arecomparatively small differences in this activity within thegroups. Although only in a few cases can a direct comparisonbe made between a cell line and tissue of origin, it appearedthat the content of CuZn Superoxide dismutase is on the wholelower in the cell lines than in the normal tissues. This has beenreported before (15, 45, 46, 48, 52, 57). The enzyme activityin the renal cancer cell line HCV ¡slower than in the kidneys.On the other hand, the activities of the various lymphoma cellswere not lower than that of the lymphocytes and the lymphaticnode but were in a few cases higher. A similar observation hasbeen reported before (66). Among the cultivated cells, lymphocytes and polymorphonuclear cells (Table 2), no general difference was seen in CuZn Superoxide dismutase activity between normal diploid cells and neoplastic cells.

There seems to be a relationship between a high metabolicactivity in tissues, and hence many mitochondria, and muchMn Superoxide dismutase; liver, kidney, heart, adrenal gland,and brain gray matter possess much enzyme (Table 1). Thereare much larger differences in Mn Superoxide dismutase activity than in CuZn Superoxide dismutase activity among cultivatedcells (Table 2). The larger differences for this enzyme may berelated to its reported inducibility (7, 11, 54, 65). Severalinvestigators have found no (45, 52) or very little (4, 46, 48,59) Mn Superoxide dismutase in various tumors and neoplastic

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Enzyme Defenses in Normal and Neoplastia Cells and Tissues

Table 1CuZn 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 dismutase

Catalase Glutathione peroxidase

LiverABErythrocytesABRenal

cortexABAdrenal

glandBRenal

medullaABSpleenABLymphatic

nodeAPancreasABLungABHeartABSkeletal

muscleABThyroid

glandABBrain

graymatterABBrain

whitematterABAdipose

tissueABU

/mg protein1,2001,5006761350470850420310952002501602601701603403302802007991540480730570200220U/g,wetwt79,0001

20,00023,00021

,00022,00037,00038,0001

6,00013,0001

1,00014,0009,2009,8009,6008,7008.60013,00013,0001

1,00011,0009,50010,0001

2,00013,0001

1,0009,6001,200800U

/mg protein31350a026404530282.27.41811305.11126673.77.20.91.216248.9139.111U/g,wetwt2,1002,900001.6003,2002,0001,1005902605106406501,0002605801,0002,7001403801101403506601302205439U

/mg protein1,3001,5009901,30043011030070022056"012010012021018054"036250011Ö3b200270560U/g,wetwt83,000114,000337,000431

,00019,3008,20012.30015,1008,2007.800604.0003,4006.3008,7006,6001

,800"01.3001,300002006100"30001.1001,800U/mg

protein190120191914087120907350200160431105354695338221512716676557789U/g,wetwt1

1,5008,9006.5006,5006,3006.4005,1002,1002,8004,8007,3005,5002,3002,4002,2002,0002.2002.0001,4001,0001,7001,2001,5001,7001,100900300280

'' 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 super-oxide dismutase is intimately related to the cancerous pheno-

types (46). This proposition was based on analysis of relativelyfew cell lines and tumors, and most investigations were madeon animal material. Our results on human cell lines (Table 2)do not support the proposition. We find highly variable amountsof Mn Superoxide dismutase in the cultivated neoplastic cellsand in some cases very large amounts. A similar finding inhuman solid tumors was recently reported (64). The difference

may be due to the fact that Mn Superoxide dismutase isapparently found in both cytosol and mitochondrial matrix inprimates (37) whereas in other species the enzyme is foundonly in mitochondrial matrix. Comparison with tissue of origincan be made only in a few cases. The cell line HCV derivedfrom a renal adenocarcinoma possessed about as much enzyme as did the kidney. The various lymphatic neoplastic cellsvaried in enzyme content but possessed about the same quantities of enzymes as did the normal lymphocytes and a little less

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Table 2

CuZn Superoxide dismutase, Mn Superoxide dismutase, catatase, and glutathione peroxidase in humannormal diploid and human neoplastic cell lines

A few of the cell lines have not been described in publications. They were taken into culture by Fogh.Roos. and Lundgren, and zur Hausen.

CelllineAnchorage

dependentFibroblastsSkin(HED21)Lung

(HEL20)AdultskinEmbryonal

lungEndothelial

cells, umbilicalcordECFibroblasts

(SV40transformed)WÕ38clone Va13Ovarian

adenocarcinoma,epithelial,aneuploidA7A10Cervical

adenocarcinomaHeLaRenal

carcinoma, epithelial, aneuploidHCV29Malignant

mesotheliomaP7(epithelialaneuploid)P31

(epithelialaneuploid)P27(fibroblastoidaneuploid)P30(fibroblastoid aneuploid)CuZn

super-oxide dismutase (U/mgprotein)1301301108318013014014016072240250200205Mn

Superoxidedismutase (U

mi iprotein)12148575472.05.3324.1241264023013Catalase(U/mgprotein)749382735.026352925084185.530NDCGlutathi

one peroxidase(U/mgprotein)4548233776469.2487039627.773NORef.2121231116abt>t>b

Giant cell tumor (fibroblastoid aneuploid)

JC1

Amnionic fluid (epithelial aneuploid)Amnion U

Glioma251 MG

Suspension growingNormal blood lymphocytes (n •21)

Normal blood polymorphonuclear leukocytes (n = 4)

Lymphoblastoid diploid B-cellsLÕA

240250200205170120120143

±30"*114

±12126

40230131.43.25.74.4

±1.42.1

±0.34185.5

30NDCNO3528NDNO62

7.773NDND30170NDNDb

b6

tb5550

160 22 62 36

Myeloma, plasmablastsRPMI8226 300

Plasma cell leukemia lymphoblastoidB-cells

IB 150

5.1

3.0

67

79

65

140

36

Burkitt's lymphoma, B-lymphoblast

aneuploidNamalwa 180

Lymphocytic leukemia T-lymphoblastJurkat 300CCRF-CEM 240Molt 4 170

3.2

8.73.41.3

32

110240110

1.5

6.15.6

ND

24

1341

Histiocytic lymphomaPlatzU 937310 2508.8 4.724 590122504456

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Enzyme Defenses in Normal and Neoplastia Cells and Tissues

Table 2—Continued

CelllineMyeloid

leukemiaHL60promyelocytesKG1myeloblastsK

562-4 blastic crisisCuZn

super-oxidedismu-tase(U/mgprotein)190220210Mn

Superoxidedismutase(U/mg

protein)3.66.83.6Catalase(U/mgprotein)92025043Glutathi-one

per-oxidase(U/mgprotein)64470Ref.82532

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 leukemiacell line, HL 60, possessed a little more Mn Superoxide dismutase than did mature normal polymorphonuclear cells. Acomparison of the normal fibroblasts and the SV40-trans-

formed fibroblast, Wi38 clone Va13, shows a decrease in MnSuperoxide dismutase in the transformed cells. This is in accordwith what has been reported before (65). The mesotheliomacell lines were exceptional. All possessed much Mn Superoxidedismutase, the P27 extremely much, almost an order of magnitude more than the richest tissue. The method for Superoxidedismutase analysis is 10 times less sensitive for Mn Superoxidedismutase than for CuZn Superoxide dismutase, which meansthat the Mn Superoxide dismutase figures should be multipliedby 10 in order to make them comparable with the CuZnSuperoxide dismutase figures. These cells are thus extremelywell equipped with Superoxide dismutases. They should forman interesting object for the study of the protective importanceof the Superoxide dismutases under various conditions. Wehave thus far not been able to obtain normal mesothelial cellsfor comparison. The mesothelioma cell lines and several of thetissues were also studied by electrophoresis and subsequentstaining for Superoxide dismutase. This was done in order tocheck the unexpectedly large amounts of Mn Superoxide dismutases found. In every case, the semiquantitative evaluationof the stained gel agreed with the presented enzymic activities.The electrophoretic mobilities of the mesothelioma CuZn su-peroxide dismutase and Mn Superoxide dismutase appearedequal to those of the normal tissues.

The content of CuZn and Mn Superoxide dismutase in thetissues shows no correlation to generally assumed radiosen-sitivities or to sensitivity to cytostatic drugs. For example,radiosensitive tissues like lung and lymphatic node containabout the same amount of Superoxide dismutase (per g, wetweight) as do less sensitive tissues like heart, skeletal muscle,and brain gray matter. This conclusion is also supported by thefindings among the cell lines. The activities of the generallyradiosensitive lymphocytic cell lines and normal blood lymphocytes are not much different from those of nonlymphocyticcells. The findings of very large amounts of Mn Superoxidedismutase in the mesothelioma cell lines are difficult to evaluatewith respect to clinical radioresponsiveness, since althoughthese tumors are considered less responsive the clinical experience of treatment is limited.

Catalase. Catalase catalyzes the disproportionate of hydrogen peroxide, 2H2O2 -» O2 + H2O, which is formed byionizing radiation and probably by the potentially radical-producing cytostatic drugs. Most tissues (Table 1) possess very

little catalase, which makes analysis with conventional methodsdifficult. This problem was overcome by the present very sensitive method. Another problem is the very high catalase content of erythrocytes which makes correction for blood contamination necessary. This was done for the tissues of the presentinvestigation. In a few blood-rich catalase-low tissues, thecorrections were so large that there was no significant remaining activity. Very large differences are found in catalase activityamong the tissues and cell lines. Most neoplastic cell lineswere low in catalase activity although some possessed largeamounts, like the promyelocytic leukemia cell line HL 60, thehistiocytic lymphoma cell line U 937, the epithelial HeLa cellline, and the acute T-lymphocytic leukemia lines Jurkat, CCRF-

CEM, 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 normalerythrocytes. No general difference between nontransformedand neoplastic cells and tissues can be seen. Several authorshave reported very low catalase activities in tumor cells (5, 61 ),although ample amounts have also been found (51 ). A correlation between catalase activity and radiation resistance hasbeen claimed (33, 61), but the general view appears to be thatthis activity is of minor importance (58). Neither is such acorrelation apparent in the present material. For example, thelymphatic tissues spleen and lymphatic node contain evenmore catalase than do "radioresistant" tissues like skeletal

muscle and brain gray matter, and the liver contains very muchcatalase (as well as the other enzymes) although it is not themost radioresistant tissue. Among the cell lines, the lymphocytic cells do not differ from nonlymphocytic cells. An interesting finding is, however, the difference in catalase and in glu-

tathione peroxidase activity of the 2 histiocytic cell lines Platzand U 937. It is a well-known clinical experience that some

histiocytic lymphomas may be highly radiosensitive while others respond more sluggishly. It is also known that differentsubpopulations of lymphocytes have different radiosensitivities,the T-helper lymphocytes being much more radioresistant thanare B-lymphocytes and T-suppressor cells (12). Our T-lympho-

blast 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 hydroperoxidesand thus has a broader protective spectrum than does catalase.Compared with catalase, there were smaller differences inglutathione peroxidase activity between the various tissues

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(Table 1) and cell lines (Table 2). The general impression isthat the activities of the neoplastia cell lines are evenly distributed within the range of the normal tissues. The results are inaccordance with previous reports of significant amounts ofglutathione peroxidase in neoplastic cells (5, 48). As for theother enzymes, there is no apparent relation between thisactivity and generally assumed resistance towards radiationand potentially radical-producing cytostatics.

Doxorubicin Toxicity and Enzyme Activity. The cardiotoxicaction of doxorubicin has been linked to toxic oxygen metabolites. An increased lipid peroxidation (43) and a decreasedglutathione peroxidase content (10) has been found in theheart after doxorubicin administration. Animals raised on aselenium-deficient diet become deficient in the selenium en

zyme glutathione peroxidase. They are much more susceptibleto doxorubicin toxicity (10). Whereas ample amounts of theSuperoxide dismutases are found in the heart (Table 1), theactivities of enzymes protecting against hydroperoxides, glutathione peroxidase and catalase, are very low. This fact maycontribute to but not provide a full explanation for the highdoxorubicin susceptibility of the heart since equally low activities are seen in skeletal muscle and in thyroid gland. Braintissue is excluded from this comparison since the drug doesnot penetrate the blood-brain barrier. Whereas a low catalase

content in mouse heart compared to mouse liver recently wasreported, almost equal glutathione peroxidase activities werefound (10). Another recent investigation of the tissue distribution of catalase and glutathione peroxidase in the mouse (18)resulted, however, in findings very similar to those presentedhere for human tissues.

Determinants of Enzyme Activities. There are considerabledifferences in the content of the enzymes in the tissues. Thismust to a large extent be due to the demands created bymetabolic specialization within the cells and to various environmental factors such as different oxygénation and exposure tovarious metabolites. The various cell lines were cultivated under comparatively similar environmental conditions created bythe nutritive cell culture medium. However, the differences inenzyme content in the cell lines appear to be as large as thedifferences between various tissues. This indicates that theinherent character and metabolic specialization of cells are themain determinants of the enzymic activities and that suchcharacters are retained in the cultivated cells.

Conclusions. Toxic oxygen reduction metabolites have beenimplicated in the damage brought about by ionizing radiation,especially in the presence of oxygen, and in the effects ofseveral cytostatic compounds. CuZn Superoxide dismutase,Mn Superoxide dismutase, catalase, and glutathione peroxidase form the primary enzymic defense against toxic oxygenreduction products. The enzymes might conceivably be ofimportance both for the response to ionizing radiation and forthe response to some cytostatics. No general difference in anyof these enzymes between normal cells and tissues and neoplastic cell lines could be demonstrated in the present investigation, nor with certainty could the activities be correlated withresistance towards ionizing radiation and potentially radical-producing drugs.

ACKNOWLEDGMENTS

We wish to thank Professor Bo Littbrand for his advice and stimulatingdiscussions. The skillful technical assistance of Margareta Bäckström,KjerstinLindmark, and Agneta Oberg is gratefully acknowledged.

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1982;42:1955-1961. Cancer Res   Stefan L. Marklund, N. Gunnar Westman, Erik Lundgren, et al.   Lines and Normal Human TissuesGlutathione Peroxidase in Normal and Neoplastic Human CellManganese-containing Superoxide Dismutase, Catalase, and Copper- and Zinc-containing Superoxide Dismutase,

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