[CANCER RESEARCH 47, 718-722, February 1, 1987]

cis-Diamminedichloroplatinum(II)-DNA Adduct Formation in Renal, Gonadal, and

Tumor Tissues of Male and Female RatsEddie Reed, Charles L. Litterst, Curtis C. Thill, Stuart H. Yuspa, and Miriam C. Poirier1

Laboratory of Cellular Carcinogenesis and Tumor Promotion, Division of Cancer Etiology [E. R., C. C. T., S. H. Y., M. C. P.] and Developmental Therapeutics Program,Division of Cancer Treatment [C. L, L,], National Cancer Institute, Bethesda, Maryland 20892

ABSTRACT

m-l)ianiniim-dichloroplatinum(II) (cisplatin), a potent anticanceragent, is thought to exert its cytotoxic effects through DNA damage.Usinga polyclonalrabbit antisera which recognizesintrastrand bidentatedeoxy(ApG)- and deo\y(Gp(;)-/V7-diammineplatinumadducts, an enzyme-linkedimmunosorbentassay has been developedto quantitate thisadduct in cisplatin-exposedDNA.Cisplatin-DNAadductsweremeasuredin renal, gonadal, and tumor (sarcoma) tissues of Sprague-Dawleyratsfollowingi.v. or i.p. administration of cisplatin. When drug was administered i.v.to animalsfedad libitum adduct levelswerehighest inkidneys,50% lower in s.c. sarcoma, and substantially lower in gonads. Underthese experimentalconditions,a large interindividualvariabilityin adductformationwas observed in renal and tumor tissues, and adduct levels insomesamples were too lowto measure. Higher valuesamongindividualswere obtained using tissues of animals fasted overnight and treated i.p.Adductlevelsfollowingi.p. injectionsof drug were higher in kidneysandgonadsof male rats than in kidneys and gonads of female rats. Analysisof tissue platinumcontentdemonstratedhigher platinumlevelsin kidneysof male rats than in kidneys of female rats, but the magnitude of thisgenderdifferencein total tissue platinumwasnot as great as that observedfor adduct formation.When the influenceof castration on adduct formation was investigated,adduct levels in kidneys of castrated females werehigher than those in sham-operated females, but adduct levelsin kidneysof the castrated male animals were not substantivelydifferent from thoseseen in sham-operated male controls. We concludethat the route of drugadministration, diet, and hormonal status of the animal are factors thatmay influencecisplatin-DNAadduct formationin the rat.

INTRODUCTION

C(s-Diamminedichloroplatinum(ll) (cisplatin) was found tohave cytotoxic activity in bacterial systems 20 years ago (1) andhas since become an effective chemotherapeutic agent beingparticularly active against testicular and ovarian malignancies(2,3). The antitumor activity of cisplatin appears to be mediatedthrough modification of DNA bases by forming multiple typesof interstrand cross-links, intrastrand adducts, DNA monoad-ducts, and DNA-protein cross-links (4-10). Together, the bidentate intrastrand deoxy(ApG)- and the deoxy(GpG)-A^-diammineplatinum adducts comprise more than 80% of totalDNA-bound platinum in LI210 and Chinese hamster ovarycells (9, 10) and are also formed in peripheral blood cell DNAof human testicular and ovarian cancer patients receiving cisplatin chemotherapy (11, 12).2 In these latter investigations,

the bidentate intrastrand adducts were quantitated by anELISA3 utilizing a polyclonal rabbit antiserum. The datashowed an accumulation of cisplatin-DNA adducts in humanDNA with increasing monthly cycles of drug treatment, whichsuggested slow removal processes and adduct persistence (12).

Received 3/31/86; revised 7/18/86, 10/16/86; accepted 10/23/86.The costs of publication of this article were defrayed in part by the payment

of page charges. This article must therefore be hereby marked advertisement inaccordance with 18 U.S.C. Section 1734 solely to indicate this fact.

1To whom requests for reprints should be addressed, at LCCTP, DCE,National Cancer Institute, NIH, Bldg 37, Room 3A23, Bethesda, MD 20892.

'!•:.Reed, S. J. Lippard, W. Sundquist, and M. C. Poirier, unpublished

observations.3The abbreviations used are: ELISA, enzyme-linked immunosorbent assay;

GSH, glutathione.

In addition, these studies indicated that two-thirds of ovariancancer patients with >200 amol of adduct/Vg DNA in theirperipheral blood cell DNA achieved a complete response tochemotherapy whereas patients who did not form adducts hada high rate of therapy failure (12, 13). Thus, high adduct levelsin human blood cell DNA are directly related to patient response. However, these studies could not be designed to learnwhat factors may play roles in the modulation of the extent ofadduct formation.

The rat has been a good model for predicting both qualitativeand quantitative toxicity of cisplatin in humans (14). In thepresent study, this model was chosen to evaluate the relationship between DNA adduct formation and tumor response andtoxicity and to analyze the relative distribution of cisplatin-DNA adducts in different tissues. In humans, kidney and gonadal tissues are targets for two major toxicities associated withcisplatin use, renal failure and sterility (15, 16). These initialstudies have focused on identifying factors which may influencecisplatin-DNA adduct formation in tumor, kidney, and gonadaltissues of the rat.

MATERIALS AND METHODS

Rats and Cisplatin Administration. Male or female Sprague-Dawleyrats (175-200 g) were housed 4-5/cage in clear plastic cages withhardwood bedding. In all but the first experiment (Table 1), animalswere fasted overnight prior to drug exposure. At all other times, theywere given free access to Purina laboratory chow and tap water. Cisplatin injections were given either i.v. or i.p. in 0.9% NaCl solution.Animals receiving different doses but treated concurrently were allgiven the same volume of solution. Injections were given i.v. with lightether anesthesia. Both i.v. and i.p. injections were given within 30 minof the time of preparation of the drug. Animals were killed by etheroverdose 4 h after injection, and kidneys, tumors, and gonads wereexcised and frozen at -20V until DNA isolation.

Studies with tumored animals were conducted with female Sprague-Dawley rats carrying a s.c. Walker 256 carcinosarcoma. Tumor stockwas maintained as an asdics in 4-6-week-old rats. When solid tumorswere needed for experiments, female rats were given injections in theflank. Tumors were palpated every second day, and when the majorityof tumors were 2-3 cm in diameter, animals with larger or smallertumors were culled and the remainder used for experiments.

Castrated and sham-operated male (6 weeks) and female (7 weeks)Sprague-Dawleyrats were obtained from Charles River Breeding Laboratories, Kingston, NY. Surgical removal of the ovarieswas performedby means of bilateral incisions through the dorsal wall of the peritonealcavity,which were closed with sutures. The testes were excised throughan incision in the scrotum, which was closed with wound clips. In bothcases, sham operations were conducted on control animals from thesame litter.

The animals were allowed to recover 6 days before shipping and wereacclimatized in our animal facility another 7 days before i.p. cisplatininjection. At the time of cisplatin injection, blood samples were takenfor analysis of testosterone, estrogen, progesterone, and lutein-stimu-lating hormone. These hormone levels were determined by routinedouble-antibody radioimmunoassays. In addition, wet weights of seminal vesiclesand uteri were recorded at autopsy.

RenalClearanceStudies. Renal clearance of platinum wasdeterminedon pentobarbital-anesthetized male and female Sprague-Dawley rats

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CISPLATIN-DNA ADDUCTS IN RAT TISSUES

Table 1 Adduci levels in cisplatin-exposed female rats bearing a s.c. Walker

Miiuiladduct/Mg DNA

( ¡splatin Tissue Rat 1 Rat 2 Rat 3 Rat 4 Rat 5 Mean ±SE

8mg/kgi.v.30

mg/kgi.p.Kidney

TumorKidney

Tumor579

252957

461118

511429

517173

851224

546ND"

22929131016704021288

117635

±360204±631038

t 202390±80

°ND, not detectable, or below 25 amol/^g DNA.

(270-300 g). Body temperature was maintained at 37 ±l'C by means

of IR heat lamps. Polyethylene cannulae were inserted into a femoralvein and artery of anesthetized animals, and a midline abdominalincision was made to permit insertion of a cannula into one ureter toallow for urine collection. The incisions were lightly closed with woundclips. After a 60-min stabilization period, 8 mg/kg of cisplatin in 0.9%NaCl solution was administered, one-third as a bolus and the remainderas a 90-min infusion (1.2 ml/h), through the venous cannula. Six urinecollections of 10 min each were then taken. At the midpoint of eachurine collection period, a 200-^1 arterial blood sample was collected ina heparinized syringe. The platinum content of the urine samples andof an uIira till rate of plasma (Centrifree filters; Amicon Corp., Danvers,MA) was determined by atomic absorption spectroscopy. Urine volumewas determined by weighing the amount of urine collected during eachperiod and assuming a specific gravity of 1.0 g/ml. Plasma was preparedfrom blood by centrifugation. Clearance (ml/min) was calculated byaveraging the quotient of platinum excreted in the urine Gig/min)divided by the plasma platinum concentration dig/nil).

DNA Preparation, ELISA, and Data Analysis. DNA was extractedfrom all tissues by CsCl gradient centrifugation (17) within 1 monthafter the tissue was harvested. The DNA samples were dialyzed againstwater for 24-36 h and the DNA was quantified by spectrophotometricabsorbance at 260 nm. Each sample was assayed for cisplatin-DNAadducts by an ELISA as described previously (7, 11). Data for 4-6animals per point are plotted as mean ±SE. Control DNAs run oneach ELISA plate were unmodified calf thymus DNA. The standardcurve consisted of calf thymus DNA of known cisplatin modification(determined by atomic absorption spectroscopy by S. J. Lippard, MIT),assayed in serial dilution, and the 50% inhibition was at 10.0 ±2.0fmol of platinum per well.

Measurement of GSH, Blood Urea Nitrogen, and Total Tissue Platinum. GSH concentrations were estimated by determining total nonpro-tein sulfhydryl content of blood and tissue. Samples were analyzedimmediately upon removal from the animal by homogenizing tissue in,or mixing blood with, ice cold 4% sulfosalicylic acid. After centrifugation at 1000 x g, the supernatant was analyzed for nonprotein sulfhy-dryls by the method of Ellman (18), using a standard curve preparedfrom reduced glutathione.

Blood urea nitrogen was determined in fresh plasma by the diacetyloxime method, using reagent kits (Sigma Chemical Company, St.Louis, MO).

Total tissue platinum concentrations were determined in homoge-nates (5 volumes of 0.25% Triton X-100 in distilled water) of tissuefragments. Homogenates were injected directly into a HGA-2100graphite furnace of a Perkin-Elmer model 403 atomic absorptionspectrophotometer. The platinum concentrations were extrapolatedfrom a standard curve prepared from hexachloroplatinic acid in distilledwater (19).

results are shown in Table 1. An overall dose-response effectwas observed with respect to adduci formation in kidney andtumor. Regardless of route, there was a large interindividualvariability in both kidney and tumor, and the ratio of tumoradduct level to kidney adduct level varied within both treatmentgroups. In almost every case, levels in kidney were higher thanin tumor. However in the 8-mg/kg group, only one value (1670amol//ig) changes the mean kidney value to being different fromthe mean tumor value. This high degree of interindividualvariability made statistical analysis of the data difficult andnecessitated the use of large numbers (5-6) of animals pergroup. A series of experiments using only normal tissues wasinitiated to determine what factors may cause the variabilityobserved.

Comparison of DNA Adduct Formation and Renal Toxicity inKidneys of Female Rats Fasted or Fed before Cisplatin Exposure.Experiments were performed to determine the effects of overnight fasting on adduct formation in the kidney. In all of threeexperiments, adduct levels were higher in kidneys of rats fastedovernight before cisplatin exposure than in renal tissue of ratsfed ad libitum. The results of a representative experiment areshown in Table 2. In this experiment the kidney platinum levels,the percentage of cisplatin dose excreted, and plasma creatininewere similar in the two groups. In addition the plasma creatinine levels were not different from the untreated controls,indicating that neither regimen resulted in a high degree ofkidney toxicity. In subsequent experiments the overnight fastingregimen was used because the overall adduct levels were higher,which maximized the probability that adducts would be measurable in tissues from all members of a group (data not shown).

Cisplatin-DNA Adduct Formation in Kidney and Gonads ofMale and Female Rats as a Function of Dose. Initially, dose-response curves for cisplatin-DNA adduct formation in kidneysand gonads were determined for male and female rats in separate experiments (Fig. I.-/). Kidney DNA adduct quantities in

males were higher at all doses than those observed in testiculartissue or in female renal or ovarian tissue. The dose-responsecurve for kidney DNA adducts was clearly nonlinear, with a 4-7-fold increase in kidney DNA adduct level associated with anincrease in dose between 20 and 30 mg/kg. The curves forkidney adduct levels, shown in Fig. 1^4,did not change significantly when expressed as semilog plots (data not shown). Inmale testicular tissues, adduct quantities were substantiallylower than in male kidney at all doses. In female ovarian tissuesadducts were lower than in female kidney, and could only bemeasured at the highest dose. Cisplatin-DNA adduct levels inmale kidneys were approximately 4-fold higher than those offemale kidney at each dose. Fig. IB shows the correspondingtotal tissue platinum values for male and female kidneys, wherethe magnitude of the male:female difference was only 2-fold.

A study was then performed in which male and female ratswere given i.p. injections from the same solution of cisplatin atdoses of 10 and 30 mg/kg body weight (Table 3). Cisplatin-DNA adduct levels determined in renal and gonadal tissues

RESULTS

Cisplatin-DNA Adduct Levels in Female Rats Bearing a s.c.Walker Sarcoma. These studies were designed to compare thelevels of cisplatin-DNA adducts in kidney and tumor tissue ofindividual tumor-bearing rats. Two experiments were performed using different routes of administration (i.v. and i.p.injections) and, in both cases, feeding of rats was ad libitum.Different doses of cisplatin were used (8 and 30 mg/kg). The

Table 2 Comparison of renal cisplatin-DNA adduct levels and kidney toxicity infemale rats fed ad libitum or fasted overnight before i.v. exposure to 8 mg/kg

cisplatin

No. ofanimals

amol adduct/MgDNA

Kidneyplatinum

Oig/g tissue)

% of cisplatin Plasmadose excreted creatinine

(4 h) (mg/dl)Fasted (5) 903 ±138" 51.4 ±4.8 24.3 ±3.5 0.4 ±0.2

Fed (5) 521 ±62 45.2 ±3.4 19.1 ±3.4 0.4 ±0.1Fasted controls (2) O O O 0.3 ±0.2" All values are expressed as mean ±SE, except the data for plasma creatinine

which are expressed as mean ±range.

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CISPLATIN-DNA ADDUCTS IN RAT TISSUES

Fig. 1. Cisplatin-DNA adduct formation inkidney and gonadal tissues ( I )and total kidneytissue platinum levels (/() in male and femalerats sacrificed 4 h subsequent to i.p. injectionsof cisplatin. I. adduct levels in kidneys (O)and testes (•)of male rats (both organs fromthe same animals). Adduct levels in kidney (A)and ovary (A) of female rats (both organs fromthe same animals). Males and females weredosed on different occasions with differentbatches of drug. H. total cisplatin tissue levelsin kidney of male (O) and female (A) rats,determined by atomic absorption spectros-copy. Samples are the same as those used inA. Each point represents mean ±SE (bars) for4-5 animals. cis-DDP, cu-diamminedichloro-platinum(II).

5000

S

O

ÕÕoo

1500

1000

500

290

200-

íS 150

g

100

50

5 10 20 30

mg c/s-DDP/Kg BODY WEIGHT

5 10 20 30

mg c/s-DOP/Kg BODY WEIGHT

Table 3 Dose-responsefor cisplatin-DNA adductformation in kidneys and gonads of male and female rats dosed simultaneously by i.p. injection

Cisplatin Kidney plat- amol adduct/Vg DNA Blood ureainum Oig/g nitrogen Renal GSH

Genderdose

(mg/kg) tissue) Kidney Gonads (mg/dl) Gig/g tissue)

FemaleMale0103001030024.8±5.0"65.9

±9.0030.0

±3.078.8±5.001522

±2063903±46802552

±5656497±4080ND*84

±330145

±34161±4922.0

±2.022.6±2.021.6±1.017.5

±2.015.0±3.017.5

±2.0766

±45771±19750±341054

±821081±301098±41

" All values are expressed as mean ±SE for 4-5 animals.* ND, not detectable or below 25 amol/Vg DNA.

were compared to total kidney tissue platinum levels. In addition, data for blood urea nitrogen were included as a controlfor kidney function, and kidney GSH levels were monitored todetermine if differences in GSH content varied with adductlevel. Adduct levels were almost 2-fold higher in kidneys ofmale rats as compared to females, whereas the total kidneyplatinum in males was only slightly higher. The magnitude ofthe adduct differences was less than in the previous experiment(Fig. 1), and overall adducts levels were higher, but the genderdifferences were again present. Male rats formed more adductin kidney tissue than female rats, even when adduct values arecorrected for differences in total tissue platinum level. Adductquantities measured in testes were higher than those observedin ovaries at both doses. Differences in blood urea nitrogen andGSH levels between male and female animals were observed,but the values for the cisplatin-treated animals were similar totheir respective untreated controls. If kidney GSH levels werea major influence on adduct formation, an inverse correlationmight be predicted between GSH and adducts; however, inthese experiments, males had more GSH and more DNAadducts than females.

At the highest dose of cisplatin, 30 mg/kg, plasma platinumvalues were consistently higher in female rats than in males(not shown). For this reason renal clearance of the drug wasstudied in six female and two male rats by the steady-stateinfusion method (20). Renal drug clearance in the female ratswas 0.39 ±0.14 nil/in in, as compared to 1.1 ±0.2 nil/in in inmale rats. The value for male rats was similar to that previouslyfound (20). Therefore, cisplatin clearance in male rats is morethan twice as rapid as in females. In summary, female rats have

slower renal clearance of cisplatin than males, show higherplasma platinum levels as compared to males, and form feweradducts in kidney DNA.

Comparison of Kidney DNA Adduct Formation in Castratedand Sham-Operated Male and Female Rats. Castrated andsham-operated male and female Sprague-Dawley rats weretreated with cisplatin from the same stock drug solution. Beforetreatment, animals were held for 2 weeks postoperatively andhormone levels were monitored before injections of 20 mg/kgof cisplatin were given i.p. The results, shown in Table 4, includecisplatin-DNA adduct formation, kidney tissue total platinumlevels, testosterone, estrogen, and progesterone levels. Althoughtestosterone was virtually absent in the castrated male rats, thequantities of cisplatin-DNA adduct in kidney tissue remainedsimilar to those in the sham-operated controls. In females,castrated animals showed somewhat higher adduct levels inkidney DNA than in the sham-operated controls, but the differences were not significant by Student's t test. Progesterone

concentrations in the castrated female rats were double that inboth groups of males, but only about 30% of that in the sham-operated females. Estrogen levels were similar in castratedmales and females and also similar in sham-operated males andfemales. Other parameters studied in this experiment includeplatinum levels in blood, and GSH levels in blood, liver, andkidney. There were no differences in these values which couldaccount for differences seen in adduct levels.

DISCUSSION

Profiles of cisplatin-DNA adduct formation and removalhave been investigated in tissues of normal, tumor-bearing, and

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CISPLATIN-DNA ADDUCTS IN RAT TISSUES

Table 4 Comparison of kidney DNA adduct formation in castrated and sham-operated male and female rats given i.p. injections of 20 mg/kg cisplatin

GenderMaleFemaleConditionSham-operated

CastratedSham-operated

CastratedKidney

amoladduct/pg

DNA1448±262*

1267±202279

±26402 ±50Kidney

platinum (¿tg/gtissue)35.2

±0.529.1±4.023.3

±3.027.1 ±3.7Tissue

wtTestosterone

(ng/ml)7.5

±1.20.05 ±0.040.9

±0.240.6 ±0.16Progesterone

(ng/ml)5.1

±0.65.3±1.127.8

±4.39.6 ±1.4Estrogen

(PR/ml)89

±5.754±3.783

±8.665 ±2.4Seminal

vesicles747

±3460 ±4Uterus287

±16106 ±5

* All values are expressed as mean •SE for 5 or 6 animals.

castrated male and female rats. These studies were designed toinvestigate cisplatin-DNA interactions in an animal that hasbeen shown to be a good model for predicting cisplatin toxicityin humans (14). The data show that the intrastrand bidentateadducts [deoxy(ApG)- and deoxyiGpGJ-A^-diammineplatinum]

are formed in kidney, gonadal, and tumor tissues of male andfemale Sprague-Dawley rats dosed with cisplatin. The highestadduct quantities were observed in the kidneys, which are aprimary site for cisplatin toxicity in both rats and humans.Adduct levels in heterologous sarcoma DNA were about one-half of those in kidney DNA in the same animals, while adductlevels in gonads were generally substantially lower than thoseobserved in kidneys. Although tumor-bearing animals were usedin our early studies, subsequent experiments were performedwith normal animals. Gonads were chosen for these studiesbecause these organs are subject to substantial toxicity in cancerpatients (16). These data show that substantial adduct quantitiesare formed in all three tissues studied. Thus, these bidentateintrastrand adducts have a wide systemic distribution, eventhough the extent of adduct formation between individuals andwithin specific tissues is variable and appears to be controlledby a number of factors. In these studies, the variability observedin absolute adduct level among different experiments performedin an identical fashion may be partially a function of the sourceof drug utilized. We have observed differences in the quantityof kidney DNA adducts when comparing identical doses ofcisplatin obtained from various sources.4

Male rats consistently formed 2- to S-fold higher quantitiesof cisplatin-DNA adducts in kidneys and gonads than femalerats. Both kidney total tissue-bound platinum and DNA adductswere suppressed in kidneys of female rats as compared to males,but they were suppressed to different extents. These differencesexisted even though plasma platinum levels were higher infemales at all doses studied and females showed slower rates ofdrug clearance by the kidney. Experiments with castrated andsham-operated male and female rats indicated that themale:female differences in adduct formation were not relatedto the presence or absence of estrogen or testosterone. However,since the progesterone levels in castrated females were 2-foldhigher than those in males and adduct levels in male kidneyswere 3-fold higher than in that of females, one could speculatethat an inhibitory influence of progesterone on the formationof DNA adducts in the kidney is possible. Identification offactors which modulate cisplatin-DNA adduct formation in rattissues will be the focus of a continuing investigation, since theability to modulate DNA adducts may lead to enhancement ofthe therapeutic index of the drug.

One goal of these studies was to search for parallels betweenadduct profiles in cancer patients and those in the rat model. Anumber of interesting parallels have emerged. Two lines ofevidence suggest that the adduct is very persistent in humancancer patients: (a) there is an accumulation of adducts in DNA

4C. L. Litterst, E. Reed, and M. C. Poirier, unpublished observations.

of nucleated peripheral blood cells with increasing dose of druggiven over a period of months (11, 12), which suggests thatadduct removal is slow and that there are long-lived adductsformed in the granulocyte precursor cells; (b) adducts have beenfound in human tissue obtained at autopsy months after thelast dose of drug was given (21). Persistence of this adduct inrat tissue has been demonstrated previously. Two weeks afteran i.v. injection of cisplatin in male rats virtually the sameadduct levels remained in kidney DNA as were found initially(11). This is in contrast to adducts from most other DNA-damaging chemicals which are substantially removed fromDNA during the first 2 weeks after dosing (22, 23).

Other parallels can be drawn between adduct profiles incancer patients and the rat animal model. In both humans andrats, the quantity of adduct formed is proportional to dose, thedistribution of adduct encompasses a variety of tissues, andthere is interindividual variation in the extent of adduct formation. Since these rats are of an inbred strain, exogenousfactors may be partially responsible. One such factor is diet,and we have shown that overnight fasting increases the adductlevels by about 2-fold.

The studies reported here constitute an initial attempt tocharacterize cisplatin-DNA adduct formation in an animalmodel. It is clear that factors such as diet, gender, and hormonalstatus can modulate the extent of adduct formation in targettissues. Other factors, presumably exerting a major influencein this area, remain to be elucidated. Future studies will continue to investigate the nature of the sex-related differences inadduct formation and will explore the relationship betweenadduct quantity, adduct removal kinetics, and tumoricidal activity. By understanding the factors which modulate adductformation in animal models we may be able to design regimensfor human cancer patients that may improve the therapeuticindex of the drug.

ACKNOWLEDGMENTS

We are grateful for the technical assistance of Elroy Patterson, NgaNguyen, Sam Eng, Huong Cao, Dung Nguyen, Martina McLane, andBrenda Phillips.

REFERENCES

1. Rosenberg, B., Van Camp, L., and Krigas, T. Inhibition of cell division inEscherichia coli by electrolysis products from a platinum electrode. Nature(Lond.), 205:698-699, 1965.

2. Einhorn, L. II. and Donohue, J. cu-Diamminedichloroplatinum, vinblastine,and bleomycin combination chemotherapy in disseminated testicular cancer.Ann. Intern. Med., 87: 293-298, 1977.

3. Young, R. C. Gynecologic malignancies. In: H. Pinedo (ed.). Cancer Chemotherapy, pp. 340-375. Amsterdam: Excerpta Medica, 1979.

4. Zwelling, L. A., Anderson, T., and Kohn, K. W. DNA-protein and DNAinterstrand cross-linking by cis- and mjn.s-platinum(II)diamminedichloridein LI210 mouse leukemia cells and its relation to cytotoxicity. Cancer Res.,39:365-369, 1979.

5. Roberts, J. J., and Pera, Jr., M. F. DNA as a target for anticancer coordination compounds. //;. S. J. Lippard (ed.), Platinum, Gold, and Other Metal

721

on May 5, 2021. © 1987 American Association for Cancer Research. cancerres.aacrjournals.org Downloaded from

CISPLATIN-DNA ADDUCTS IN RAT TISSUES

Chemotherapeutic Agents, pp. 3-25. Washington, DC: American ChemicalSociety, 1983.

6. Plooy, A. C. M , van Dijk, M., and Lohman, P. H. M. Induction and repairof DNA cross-links in Chinese hamster ovary cells treated with variousplatinum coordination compounds, in relation to platinum binding to DNA,cytotoxicity, mutagenicity, and antitumor activity. Cancer Res., 44: 2043-2051, 1984.

7. Poirier, M. C., Lippard, S. .1.. Zwelling, L. A., Ushay, M., Kerrigan, D.,Thill, C. C, Santella, R. M., Grunberger, D., and Yuspa, S. H. Antibodieselicited against cú-diamminedichloroplatinum (II)-DNA adducts formed invivo and in vitro. Proc. Nati. Acad. Sci. USA, 79:6443-6447, 1982.

8. Lippard, S. J., Ushay, H. M., Merkel, C. M., and Poirier, M. C. Use ofantibodies to probe the stereochemistry of antitumor platinum drug bindingto DNA. Biochemistry 22: 5165-5168, 1983.

9. Plooy, A. C. M., Fichtinger-Schepman, A. M. J., Schutte, H. H., van Dijk,M., and Lohman, P. H. M. The quantitative detection of various Pt-DNA-adducts in Chinese hamster ovary cells treated with cisplatin: application ofimmunochemical techniques. Carcinogenesis (Lond.), 6: 561-566, 1985.

10. Eastman, A. Multiple mechanisms of ru-diamminedichloroplatinum(II) resistance in a murine leukemia 1.1210 line. Proc. Am. Assoc. Cancer Res., 27:291, 1986.

11. Poirier, M. C., Reed, E., Zwelling, L. A., Ozols, R. F., Litterst, C. L., andYuspa, S. H. The use of polyclonal antibodies to quantitate nVdiamminedichloroplatinum (II) ON \ adducts in cancer patients and animal models.Environ. Health Perspect., 62:89-94, 1985.

12. Reed, E., Yuspa, S. II., Zwelling, L. A., Ozols, R. F., and Poirier, M. C.Quantitäten of cisplatin-DNA-intrastrand adducts in testicular and ovariancancer patients receiving cisplatin chemotherapy. J. Clin. Invest., 77: 545-550, 1986.

13. Poirier, M. C., Reed, E., Ozols, R. F., and Yuspa, S. H. DNA adduciformation and removal in human cancer patients. In: C. C. Harris (ed.),

Biochemical and Molecular Epidemiology of Cancer, pp. 303-311. NewYork: Alan R. Liss, 1986.

14. Guarino, A. M., Miller, D. S., Arnold, S. Y., Pritchard, J. B., Davis, R. D.,Urbanek, M. A., Miller, T. J., and Litterst, C. L. Platinate toxicity: past,present, and prospects. Cancer Treat. Rep., 63:1475-1483,1979.

15. Ozols. R. F., Corden, B. J., Jacob, J., Wesley, M. N., Ostchega, Y., andYoung, R. C. High-dose cisplatin in hypertoxic saline. Ann. Intern. Mai..¡00:19-24, 1984.

16. Drasga, R. E., Einhorn, L. H., Williams, D. S., Patel, D. N., and Stevens, E.E. Fertility after chemotherapy for testicular cancer. J. Clin. Oncol., 1:179-183, 1983.

17. Flamm, W. G., Binisi id. M. L., and Walker, P. M. B. Preparation andfractionation, and isolation of single strands of DNA by isopycnic ultracen-trifugation in fixed-angle rotors. In: G. Birnie and S. M. Fox (eds.), Subcellular Compounds: Preparation and Fractionation, pp. 129-155. London:Butterworth, 1967.

18. Fllniiin. G. L. Tissue MIHin dry 1groups. Arch. Hint hem. Biophys., 82: 76-77, 1959.

19. Vozumi, J., and Litterst, C. L. Effect of cisplatin on renal ATPases in vivoand in vitro. Cancer Chemother. Pharmacol., in press, 1986.

20. Osman, N. M., and Litterst, C. L. Effect of probenecid and W-methylnico-tinamide on renal handling of cu-dichlorodiammineplatinum-II in rats. Cancer Lett., 19:107-111, 1983.

21. Reed, E., Ozols, R. F., Fasy, T., Yuspa, S. H., and Poirier, M. C. Biomonitoring of cisplatin-DNA adducts in cancer patients receiving cisplatin chemotherapy. In: C. Ramel, B. Lambert, J. Magnusson (eds.). Genetic Toxicologyof Environmental Chemicals, Pact B: Genetic Effects and Applied Mutagen-esis, pp. 247-252. New York: Alan R. Liss, 1986.

22. Beland, F. A., and Kadlubar, F. F. Formation and persistence of arylamineDNA adducts in vivo. Environ. Health Perspect., 62:19-30, 1985.

23. Stowers, S. J., and Anderson, M. W. Formation and persistence of benzo(a)-pyrene metabolite-DNA adducts. Environ. Health Perspect., 62:31-19,1985.

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1987;47:718-722. Cancer Res   Eddie Reed, Charles L. Litterst, Curtis C. Thill, et al.   Renal, Gonadal, and Tumor Tissues of Male and Female Rats

-Diamminedichloroplatinum(II)-DNA Adduct Formation incis

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