Enhancement by Organosulfur Compounds from Garlic and Onions

Home , Diallyl disulfide

[CANCER RESEARCH54, 2895-2899, June 1, 1994@ Enhancement by Organosulfur Compounds from Garlic and Onions of Diethylnitrosamine-induced Glutathione S@TransferasePositive Foci in the Rat Liver' NobuyasuTakada,2TsutomuMatsuda,TatsuyukiOtoshi,YoshihisaYano, ShuzoOtani,TohruHasegawa,Dai Nakae, Yoichi Konishi, and Shoji Fukushinia

First Department of Pathology (N. T., T. M., 7'. 0., S. F.] and Second Department of Biochemistry [V. Y, S. 0.], Osaka City University Medical Schoo4 Asahi-machi Abeno-kis, Osaka 545, Japan; Department of Community Health Science, Saga Medical Schoo4 Saga 849, Japan [T. H.]; and Department of Oncological Pathology, Cancer Center, Nara Medical University, Nara 634, Japan ID. N., Y. K]

ABSTRACT cancer, esophageal cancer, pulmonary adenoma, and forestomach tumors in rodents when administered prior to administration of several Ten organosulfur compounds from garlic and onions were studied for carcinogens (13â€”16).Moreover, DAS inhibited hepatocarcinogenesis their modifying effects on diethylnitrosamine-induced neoplasia ofliver in when administered after an initiating procedure, but this has not been male F344 rats using the medium-term bioassay system oflto based on the two-step model of hepatocarcinogenesis. Carcinogenic potential was consistently observed (17, 18). scored by comparing the number and area per cm2 ofinduced glutathione DAS, diallyl disulfide, diallyl trisulfide, allyl methyl sulfide, allyl S-transferase placental form-positive foci in the liver with those of the methyl trisulfide, and propylene sulfide are richly contained in garlic; corresponding control group given diethylaitrosamine alone. In experi and dipropyl sulfide, dipropyl disulfide, methyl propyl disulfide, and meats 1 and 2, high doses of diallyl sulfide, diallyl trisulfide, allyl methyl dimethyl disuffide are contained abundantly in onion (7, 11, 19, 20). sulfide, allyl methyl trisulfide, and dipropyl sulfide had enhancing effects In the present study, the modifying potential in the promoting stage on focus formation. In contrast, high doses ofmethyl propyl disulfide and of ten OSCS contained in garlic and onion was examined in a rat liver propylene sulfide significantly decreased the number of glutathione S medium-term bioassay system as defined by Ito et aL (5). Moreover, transferase placental form-positive foci. In the third experiment, corn the enhancement by combined treatment with OSCS was evaluated in biasedtreatment with the five chemicals that had enhancing activity were the same system. To begin investigating possible mechanisms that fed at low doses and increased the induction of glutathione S-transferase placental form-positive foci. To investigate the mechanism of the modify might be involved, formation of 8-OHdGuo, a DNA adduct generated big effect on hepatocarcinogenesis, ornithine decarboxylase activity was by activated oxygen species, and ThA-reacting substances produced measured In diallyl sulfide-, allyl methyl sulfide-, and dipropyl sulfide in rat liver were measured (21, 22). In addition, ODC and SAT, which treated liver tissue without prior initiation with diethylnitrosarnine, and are rate-limiting enzymes of polyamine biosynthesis and were re its activity was increased compared to controk Spermidine/spermine ported to be increased in the promoting stage of skin and bladder N'-acetyltransferase activity was not significantly changed. Formation of carcinogenesis (23, 24), were also measured. 8-hydroxydeoxyguanosine,aDNAadduct generated by activated oxygen species, and lipid peroxidation (2-thiobarbituric acid-reacting substance MATERIALS AND METhODS production) were also not changed. These results suggest that the promot lag effect could be caused by increased cell proliferation with increased Chemicals. Organosulfur compounds used are DAS, DDS, DAT, AMS, polyamine biosynthesis. In evaluating relationships between diet and can AMT, DPS, DPD, MPD, PS; and DMD (19, 20). DAT and AMT were obtained car, it is appropriate to consider not only the possible protective role of from Oxford Chemicals Co., Northhamptonshire, United Kingdom; MPD was garlic and onions but also their enhancing effects. from Aldrich Chemical Co., Milwaukee, WI; and the other OSCs and DEN from Tokyo Chemical industry Co., Tokyo, Japan. Animals and Treatments. Male F344 rats obtained at 5 weeks of age INTRODUCTION (Charles River Japan, Inc., Hino, Japan) were housed in an air-conditioned room at 23 Â±1Â°C(SD),a relative humidity of 36 Â±6%,with a 12-h light/12-h Environmental compounds are known to be involved in the gener dark cycle, and were given diet (Oriental MF; Oriental Yeast Co., Tokyo, ation of many human cancers. Elimination of carcinogenic corn Japan) and tap water ad libitum. The diet is a commercial diet, which is a pounds from our environment would be expected to help prevent modification of the NIH open formula diet (NIH-07). Rats were acclimatized human cancer, but this is not a practical proposition. Therefore, it is for 1 week before use. important to discover naturally occurring or synthetic compounds Experiments 1 and 2 were performed to investigate modifying effects of which can suppress or prevent the process of carcinogenesis (1â€”12). each OSC on hepatocarcinogenesis. In experiment 1, a total of 150 rats were We have focused on some OSCs3 which are contained in garlic and divided into 12 groups. The rats in groups 1 to 5 were given a single i.p. onion. Some OSCS have been proved to be chemopreventive in animal injection of DEN (200 mg/kg body weight) dissolved in saline to initiate hepatocarcinogenesis. After 2 weeks on basal diet, they received DAS (100 models. For example, diallyl sulfide inhibits development of colon mg/kg body weight; group 1), DDS (25 mg/kg body weight; group 2), AMS (150 mg/kg body weight; group 3), DPS (150 mg/kg body weight; group 4), Received8/18/93;accepted4/5/94. and DPD (150 mg/kg body weight; group 5) dissolved in corn oil (1 mI/kg) by The costs of publication of this article were defrayed in part by the payment of page i.g. gavage 5 times/week for 6 weeks. Animals were subjected to PH at week charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. 3 to maximize any interaction between proliferation and the effects of the I This study was supported by Grant-in-Aid of Cancer Research from the Ministry of compounds tested. Group 6 was given DEN and PH without administration of Health and Welfare. any test compounds. Animals in groups 7 to 11 received saline instead of DEN 2 To whom requests for reprints should be addressed. solution but were subjected to administration of test compounds and PH. 3 The abbreviations used are: OSCs, organosulfur compounds; DAS, diallyl sulfide; DDS, diallyl disulfide; DAT, diallyl trisulfide; AMS, allyl methyl sulfide; AMD, allyl Group 12 animals were given saline injections and then subjected to admin methyl disulfide; AMT, allyl methyl trisulfide; PS, propylene sulfide; DPS, dipropyl istration of corn oil instead of test compounds; they were also subjected to PH. sulfide; DPD, dipropyl disulflde; MPD, methyl propyl disulfide; DMD, dimethyl disul Rats in each group were killed for examination at week 8 (Fig. 1). The livers fide; DEN, N-diethylnitrosamine; 8-OHdGuo, 8-hydroxydeoxyguanosine; ODC, ornithine were examined immunohistochemically for GST-P expression. decarboxylase; SAT, spermidine/spermine N1-acetyltransferase; TBA, 2-thiobarbituric acid; i.g., intragastrically; PH, two-thirds partial hepatectomy; GST-P, glutathione 5- In experiment 2, a total of 150 rats were divided into 12 groups. Rats in transferase placental form. each group were treated by the same medium-term bioassay system as 2895

@3 8weeks Groups

1â€”5 I Fig. 1. Liver medium-term bioassay protocol. V, @ DEN, 200 mg/kg body weight (i.p.); V, saline, 5 I @iiIIâ€”@â€”I mI/kg body weight (i.p.), â€¢,partial hepatectomy; 5, 6 sacrifice; @,testchemicals (i.g.) 5 times/week; 0, corn oil, 1 mI/kg body weight (i.g.). 7 @.11 [

12:______I

experiment 1. They received DAT (150 mg/kg body weight; group 1), AMT Measurement of SAT and ODC Activity and Polyamine Biosynthesis. (100 mg/kg body weight; group 2), MPD (100 mg/kg body weight; group 3), We measured SAT and ODC activity by the method of Matsui et aL (36) and PS (50 mg/kg body weight; group 4), and DMD (50 mg/kg body weight; group Otani et a!. (37), respectively. The frozen piece of rat liver was suspended in 5) dissolved in corn oil by i.g. gavage 5 times/week. All rats were killed at 0.5 ml of 50 mM Tris (pH 7.5) containing 0.25 M sucrose and disrupted by a week 8, and livers were examined immunohistochemically for GST-P expres homogenizer for 30 seconds. The homogenized suspensions were centrifuged sion, formation of 8-OHdGuo, and production of ThA-reacting substances. at 100,000 x g for 30 mm, and the supernatant was assayed for ODC and SAT The dose of DAS and DDS were one-half of the doses (200 and 50 mg/kg activity by measurement of the amount of radioactive putrescine produced body weight) used in a previous experiment by us (18). The doses of the other from [5-'4C]ornithine and the amount of acetyl moiety transferred from chemicals tested were the same or nearly equimolar concentration as DAS or [1.'4C]acetyl-CoA to spermidine, respectively. DDS. Each dose is less than the maximum tolerated dose. Statistical Analysis. Statistical analysis of the observed values was per In experiment 3, the modifying effects with combined treatment of OSCs on formed using Student's t test or Dunnett's t test. rat hepatocarcinogenesis modification were examined. A total of 105 rats were divided into 7 groups. Rats in each group were given DEN and PH with or RESULTS without combined treatment of 5 test compounds. The test compounds which increased the areas and numbers of GST-P-positive foci from experiments 1 In DEN-initiated groups of experiments 1 and 2, final body weights and 2 were DAS, DAT, AMS, AMT, and DPS. The doses of the combined were significantly decreased in rats treated with DPD, DAT, and PS treatment of five OSCs are shown in Table 1. The determination of doses were compared to rats treated without test chemicals. Relative liver weights based on the ratio of their content in garlic (groups 1â€”3)orusing equimolar were significantly increased in DEN-initiated rats treated with DDS, concentrations (groups 4â€”6). For these two combinations, three dose levels were used. In experiment 4, ODC and SAT activity were measured in the liver follow ing simple administration of the OSCs. A total of 20 rats were divided into 4 differentsynergisticcombinationsDoseGroup(mg/kg)DATAMTDASAMSDPS112032122221/2ofgroupl601661131/4Table 1 Treatment of 0SCs with two groups. From the beginning of the experiment, the rats received DAS (100 mg/kg body weight; group 1), AMS (150 mg/kg body weight; group 2), or DPS (150 mg/kg body weight; group 3) in corn oil (1 mI/kg) by i.g. gavage 5 times/week. Group 4 animals received corn oil alone. All rats were killed at week 3. I30830.50.54302020303051/2ofgroup4151010151561/4of group Tissue Processing. At autopsy, livers were excised and sections 2â€”3mm thick were cut with a razor blade. Three slices, one each from the right posterior, anterior, and caudate lobes, were fixed in ice cold acetone for 47.5557.57.5700000of group immunohistochemical examination of GST-P in experiments 1â€”3.To measure the formation of 8-OHdGuo and lipid peroxidation of rat liver in experiment 2 and ODC and SAT activity of rat liver in experiment 4, rat livers were frozen by liquid nitrogen. GST-P Immunohistochernistry. The avidin-biotin-peroxidase complex Table 2Final body and liver weights of rats initiated with DEN in experiment 1 OSCsTestfollowed by treatment with various method was used to demonstrate GST-P-positive liver foci, a putative preneo plastic lesion (5, 25, 26). After deparaffinization, liver sections were treated wtGroupchemical EffectiveRelative liver sequentially with normal goat serum, rabbit anti-GST-P (1:8000) (provided by wt)1 no. of rats Final wi (g)(g/100 g body Dr. K. Sato, Hirosaki University) biotin-labeled goat anti-rabbit IgG (1:400) 14 273 Â±13â€• Â±0.20 and avidin-biotin-peroxidase complex. The sites of peroxidase binding were 014b32DAS DDS 14 263Â±14333 3.65Â± 14 268Â±19 demonstrated by the diaminobenzidine method. Sections were then counter 4 DPS 14 268Â±13 3.92Â±O.2l@' stained with hematoxylin for microscopic examination. As a negative control 018b6145AMS DPD 14 243 Â±19â€•3.32Â±0.163.04 Â± for the specificity of anti-GST-P antibody binding, preimmune rabbit serum 274 Â±143.25 Â±0.17 was used instead of antiserum. The numbers and the areas of GST-P-positive a Mean Â± SD. @ foci >0.2 mm in diameter and the total areas of the liver sections examined b < 0.01 (versus group 6, Student's t test). were measured using a color video image processor (VIP-21C). Determination of 8-OHdGuo Formation in DNA. Liver sample portions Table 3Final body and liver weights of initiated with DEN in experiment 2 â€”2g, wet weight, were used. Liver DNA was isolated and digested as OSCsTestfollowed by treatmentratswith various described by Takagi et aL (27). The level of 8-OHdGuo formation in resulting wtGroupchemical EffectiveRelative liver samples was determined by high performance liquid chromatography by an wt)1DAT no. of ratsFinal wt (g)(g/100 g body adaptation of the methods of Floyd et a!. (28, 29) and Kasai et a!. (30â€”32)as 016b2AMT 15232 Â±14a.b3.36 Â± described previously (33). The level of 8-OHdGuo formation was expressed as Â±3MPD 14256 Â±103.04 the number of 8-OHdGuo residues formed/10@ of total deoxyguanosine. 0.18c4PS 15261 Â±122.69 Â± Determination of Lipid Peroxidation (TBA-reacting Substance Produc 0.I4'@5DMD 14245 Â±12b2.81 Â± 0.23c615261 14262 Â± tion). Peroxidationof liver lipidswas quantitatedbymeasuringaccumulation Â±102.74 Â±142.56 Â±0.17 of TBA (Sigma)-reacting substances and expressed as nmol value equivalent aMean Â± SD. @ (eq) to a standard amount of malondialdehyde (dimethyl acetal; Aldrich) by an b < o.oi (versus group 6, Student's t test). adaptation of the method of Yagi (34) as described previously (35). Cp < 0.05 (versus group 6, Student's t test). 2896

DPS, DAT, AMT, MPD, PS, and DMD and decreased in DPD-treated Table 6 Fina1 body and liver weights of rats initiated with DEN in experiment 3 group (Tables 2 and 3). OSCsEffectiveRelativefollowed by treatment with various Tables 4 and 5 show the numbers and areas of GST-P-positive foci WIGroupno. liver per unit area of liver sections after DEN initiation. In experiment 1, WI)1 of rats Final WI (g)(g/l00 g body values for both parameters in groups given DAS, AMS, and DPS were zs6Â±lla.@@ 2 14 268Â±9 3.77Â±0.10â€•' significantly increased over control levels. In particular, DPS was 3 14 266Â± 11 3.60Â±0.l6@'' most effective. In experiment 2, in groups given DAT or AMT, the 4 14 263Â±11â€• 3.69Â±0.12â€• 5 14 269Â±11 3.59Â±0.21â€• number and area of GST-P-positive foci were significantly increased. 6133.47Â±0.17â€•â€•71514 278Â± 103.94Â±0.17â€• In contrast, in groups given MPD or PS, values for the number of foci 287 Â±132.91 Â±0.16

were significantly decreased. In experiments 1 and 2, GST-P-positive a Mean Â± SD. @ foci were not seen, and the histological findings of livers were normal b < o.oi (versus group 7, Student's t test). Cp < o.os (versus group 1, Student's t test). @ in groups without DEN. d < 0.01 (versus group 4, Student's t test). For experiment 3, final body and relative liver weights are sum marized in Table 6. Final body weights were significantly decreased in rats treated concomitantly with high doses oftest chemicals (groups Number (treated/control ratIo) Area (treated/control ratIo) 1 and 4). Relative liver weights were significantly increased in all of the treated groups, and a dose dependency could be observed. The proportion of numbers and areas of GST-P-positive foci in the differ 2

ent groups compared to group 7 (control) are summarized in Fig. 2. 3 They were significantly increased by the combined treatments corn 4 pared to the control group. In each group treated with test chemicals, no significant differences were observed, but there was a tendency 5 toward a dose-dependent response. 6

Results for determinations of 8-OHdGuo formation in DNA and 7 ThA-reacting substances produced in the livers of rats initiated by :::j , I I 3 2 1 u 0 1 2 3 DEN in experiment 2 are shown in Table 7. Formation of 8-OHdGuo Fig. 2. Numbers and areas ofGST-P-positive foci in the liver ofrats initiated with DEN tended to decrease in groups treated with MPD or PS. Results for followed by treatment with various OSCs. Significantly different from group 7 at @ ThA-reacting substances displayed significant decreases in rat livers *P < 0.05, **@ < Dunnett's t test. with MPD and PS treatment. In groups treated with DAT or AMT, formation of 8-OHdGuo and ThA-reacting substances hardly in creased compared to control group. DISCUSSION For experiment 4, the results of ODC and SAT activity in the liver are shown in Table 8. ODC activity increased in DAS (42.9 Â± 12.5 Although most OSCs have been shown to be chemopreventive in pmol/mg)-, AMS (137.5 Â± 157.6 pmol/mg)-, and DPS (38.7 Â± previous studies, high doses of DAS, DAT, AMS, AMT, and DPS 34.5 pmol/mg)-treated rats over the control level (13.5 Â± 6.6 pmol/ were shown to promote preneoplastic lesion development in the rat mg) and was especially elevated significantly in the group treated with liver in the present study. Their combined treatment enhanced GST DAS. SAT activity was not increased in any of the groups. P-positive focus development. Their effects indicated a remarkable dose dependency in the two different combined groups. Therefore, their combined treatment may exert promoting activities on hepato Table 4 N and areas of GST-P positive foci in the liver of rats initiated with carcinogenesis of rats. DumbersEN inOSCsTest experiment 1 followed by treatment with various In previous studies, the inhibiting effects of OSCS were mainly No.Groupchemical examined in the initiation stage (13, 38â€”42).AMT, AMD, DAT, and (mm2/cm2)IDAS of rats No/cm2 Area DAS administered 96 and 48 h prior to the carcinogen inhibited 0.17'2DDS 14 6.58 Â±2.12'@b 0.52 Â± 0.173AMS 14 4.39 Â±1.81 0.33 Â± benzopyrene-induced tumors of the forestomach in female A/i mice. 018b4DPS 14 6.14 Â±1.71c 0.56 Â± DAS and AMD inhibited pulmonary adenoma formation in female 0.29â€•5DPD 14 8.45 Â±2.52â€• 0.88 Â± A/i mice, but neither DAT nor AMT did (41). In those experiments, 0.20614 14 5.53Â±1.39 0.43Â± 4.69 Â±1.61 0.38 Â±0.16 the administration of OSCs during the initiation stage inhibited the aMean Â±SD. metabolic activation of the carcinogens (43, 44); but in our experi b Significantly different from group 6 at P < 0.05 (Student's t test). ment, since we administered OSCs during the promotion stage, our C Significantly different from group 6 at P < 0.01 (Student's t test). results imply the involvement of a different mechanism. d Significantly different from group 6 at P < 0.001 (Student's t test). The selection of the bioassay system in this study was based on examining the modifying potential of OSCs on the promoting stage of Table 5 Nit and areas of GST-P positive foci in the liver of rats initiated with hepatocarcinogenesis. The data obtained from the present investiga OSCsGroupTestDmbersEN in experiment 2 followed by treatment with various tions unexpectedly showed enhancing effects of five OSCs, DAS, No. DAT, AMS, AMT, and DPS, several of which inhibited carcinogen (mmicm2)1DATchemical of rats No./cm2 Area esis by administration during the initiation stage. This bioassay system Â±2 15 6.62 Â±1.89â€• 0.58 has been utilized to detect modifying effects of many chemicals on 14 8.24 Â±2.71c 0.59 Â±0.15' liver carcinogenesis (5). Moreover Ogiso et a!. (45, 46) indicated that 3 MPD 15 3.28 Â±155b 0.26 Â±0.17 4AMT0.175DMD PS 14 3.17 Â±172b 0.26 Â± the degree of induction of GST-P-positive foci and nodules in this 0.16615 14 4.09 Â±1.81 0.35 Â± bioassay system for liver carcinogens corresponds with the incidence 4.71 Â±1.38 0.35 Â±0.14 of hepatocellular carcinomas revealed in a long-term in vivo assay. aMean Â± SD. Consequently the present study strongly suggests that these OSCs b Significantly different from group 6 at P < 0.05 (Student's t test). C Significantly different from group 6 at P < 0.01 (Student's t test). promote hepatocarcinogenesis. 2897

Table 7 Induction of oxidative DNA damage and ThA-reacting substance production 8-OHdGuo correlates with development of putative preneoplastic in rat livers initiated with DEN followed by treatment with various OSCs â€˜y-glutamyltransferase-positive focal lesions using a rat hepatocarci substance nogenesis model by a choline-deficient L-amlno acid-defined diet. production (@mol Takahashi et aL (55) reported that lipid peroxidation was increased in GroupTestmg)1DAT4.5Â±0.9â€•8.7Â±chemical8-OHdGuo/10@deoxyguanosineThA-reactingmalondialdehyde/l00 gastric mucosa by NaCl, a stomach cancer promoter. That study 1.52 Â±0.8 clearly demonstrated that the gastric tumor promoter NaCl dose 3 MPD 3.1 Â±1.4 8.0Â±3.0â€• dependently increased ThA-reacting substances production levels in 6.9Â±l.l@'5DMD4.7Â±2.08.2Â±4AMT PS4.1 3.6Â±1.59.4Â±0.5 1.164.7 both gastric mucosa and urine, supporting a conclusion that there is Â±2.211.2 Â±1.8 enhanced lipid peroxidation in the target tissue. Although increased aMean Â±SD. formation of 8-OHdGuo in DNA and lipid peroxidation were cx b Significantly different from group 6 at P < 0.05 (Student's t test). pected to participate in the promoting effects of OSCs, they were not significantly involved as determined in the present study. Table 8 ODC and SAT activity in liver of rats treated with OSCs In this study, the number but not the area of GST-P-positive foci Test activity activity was significantly decreased in rats given PS or MPD. This suggests chemicalODCprotein)DAS42.9 (pmol/hlmg protein)SAT (pmol/10 min/mg that these compounds might have inhibitory effects on hepatocarci 1.20AMS137.5 Â±12.5a0.24 Â± 0.90DPS38.7 Â±1576b0.82 Â± nogenesis. In addition, the reason why induction of 8-OHdGuo and Â±34.5 Â±0.72 lipid peroxidation were decreased in the liver should be clarified with 13.5 Â±6.60.45 0.77 Â±1.42 further experiments. aM@ Â± SD. The test chemicals are sulfur-containing, volatile compounds, and b Significantly different from the nontreated group at P < 0.05 (Student's t test). especially the monosulfides, except PS, and the trisulfides have a tendency to enhance GST-P-positive focus formation. The number of Most of OSCs used may tend to decrease body weights and increase sulfur atoms may relate to the mechanism of promotion of hepatocar relative liver weights. In further studies, we must examine the dose cinogenesis by OSCS (56). response for each OSC which enhanced GST-P positive focus forma Humans are exposed to numerous environmental chemicals during tion in order to evaluate the possibility of a practical threshold. In their life span and the chemicals may act in combination, positively or addition, we must dissociate the effects of enhancing focus formation antagonistically, to affect cancer production. Moreover synergistic from the toxicity of OSCS. effects of chemicals on development of tumors have been repeatedly A modifying response of DAS in the promoting stage was exam observed in many organs (57, 58). We observed a dose-dependent med in a rat multiorgan carcinogenesis model by Jang et a!. (17) and tendency in the promoting effects by combined treatment with five Takahashi et a!. (18), with different results. Jang et al. reported that OSCs (DAS, DAT, AMS, AMT, and DPS) in two different combi DAS administered in the diet significantly decreased the incidence of nation groups. Since we also found two inhibitors, we need additional hepatic hyperplastic nodules in the liver. However, Takahashi et a!. studies to examine the effects of combined treatment with enhancers reported that DAS demonstrated clear enhancing effects on hepato and inhibitors. It is possible that low doses of carcinogenic substances carcinogenesis. In the experiments of Takahashi et aL and in our in combination are of critical importance in determining effects. This study, rats received DAS by i.g. intubation, which was quite different may contribute to the correlation between hepatocarcinogenesis and from the method of Jang et a!. Moreover, Takahashi et a!. reconfirmed intake of garlic and onion. their results in the DEN-initiated liver medium-term bioassay con In conclusion, many OSCs which exerted promoting effects in cerning promoting effects of DAS. Our present findings are in agree hepatocarcinogenesis exist in garlic and onion. ODC activity in ment with the latter. The reason for this discrepancy with Jang et a!. creased in the treated groups. Moreover, these data suggest that is unclear. cell proliferation in liver tissue is important for the promoting Cell proliferation has long been suspected of enhancing the fre effects of OSCs. quency of tumor initiation in chemical carcinogenesis. In addition, cell proliferation appears to play an important role in the effects of tumor promoters or nongenotoxic carcinogens (47, 48). Polyamines ACKNOWLEDGMENTS are involved in epithelial cell proliferation (49). ODC, which is a We extend our gratitude to the late Professor Kiyomi Sato of the Second biomarker of cell proliferation, increased with epithelial cell prolifer Department of Biochemistry, Hirosaki University, for the generous provision ation of skin or bladder when a promoting agent was administered (23, of purified antibody against GST-P. 24). ODC activity is high in carcinomas induced experimentally by chemical carcinogens (50, 51) and in carcinomas obtained from pa tients (52, 53). SAT is a newly established rate-limiting enzyme of REFERENCES biodegradation of polyamines (36) and it also was found to be a 1. Ames, B. N. Dietary carcinogens and anticarcinogens. Science (Washington DC), biochemical marker of cell proliferation (49). 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Downloaded from cancerres.aacrjournals.org on September 27, 2021. © 1994 American Association for Cancer Research. Enhancement by Organosulfur Compounds from Garlic and Onions of Diethylnitrosamine-induced Glutathione S -Transferase Positive Foci in the Rat Liver

Nobuyasu Takada, Tsutomu Matsuda, Tatsuyuki Otoshi, et al.

Cancer Res 1994;54:2895-2899.

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