Exceptional Activity of Tannic Acid Among Naturally Occurring Plant

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(CANCER RESEARCH 48, 2361-2365, May 1, 1988) Exceptional Activity of Tannic Acid among Naturally Occurring Plant Phenols in Protecting against 7,12-Dimethylbenz(a)anthracene-, Benzo(a)pyrene-, 3-Methylcholanthrene-, and TV-Methyl-TV-nitrosourea-induced Skin Tumorigenesis in Mice Hasan Mukhtar,1 Mukul Das,2 Wasiuddin A. Khan, Zhi Y. Wang, Daniel P. Bik, and David R. Bickers

Departments of Dermatology, University Hospitals of Cleveland and Case Western Reserve University, and the Veterans Administration Medical Center, Cleveland, Ohio 44106

ABSTRACT cells (8,9,12). Our recent studies have shown that plant phenols such as tannic acid, quercetin, myricetin, and anthraflavic acid Our recent studies nave shown that naturally occurring dietary plant were potent inhibitors of epidermal monooxygenases, BP me phenols such as tannic acid, quern-tin, myricetin, and anthraflavic acid tabolism, and the binding of [3H]BP, [3H]BP-7,8-diol, and are capable of inhibiting polycyclic aromatic hydrocarbon (PAH) metab [3H]-DMBA to epidermal DNA in SENCAR mice (13, 14). olism and subsequent PAH-DNA adduci formation in epidermis of SENCAR mice (M. Das, et al.. Cancer Res., 47:760-766,1987, and 47: Our studies also indicate that topical application of each of 767-773, 1987). In this study these plant phenols were tested for their these plant phenols inhibits in vivo epidermal and pulmonary effects against PAHs and yV-methylWV-nitrosourea-induced skin tumori- BPDE-I-deoxyguanosine adduct formation in [3H]BP-treated genesis in mice. Each plant phenol was evaluated as a possible anticar- SENCAR mice (14). It was therefore of interest to evaluate the cinogen in an initiation and promotion and a complete skin tumorigenesis ant Â¡carcinogenicactivity of these polyphenols. We report that protocol. In the two-stage tumor protocol in SENCAR mice using 7,12- tannic acid, quercetin, myricetin, and anthraflavic acid also dimethylbenz(a)anthracene, benzo(a)pyrene, and JV-methyl-./V-nitroso- exhibit variable degrees of protection against DMBA-, BP-, urea as the initiating agent followed by twice weekly applications of 12- MCA-, and MNU-induced skin tumorigenesis in mice. O-tetradecanoylphorbol-13-acetate as tumor promoter each plant phenol afforded significant protection against skin tumorigenicity. The protective effects were verified both by prolongation of latency period and by MATERIALS AND METHODS subsequent tumor development. In the complete carcinogenesis protocol in BALB/c mice using 3-methylcholanthrene as a tumorigen the appli Chemicals. Tannic acid, myricetin, anthraflavic acid, DMBA, and cations of each of the plant phenols 30 min prior to each PAH application BP were obtained from Aldrich Chemical Co., Milwaukee, WI. Quer afforded significant protection by delaying the onset and the subsequent cetin, MCA, and TPA were purchased from Sigma Chemical Co., St. development of skin tumors. Our results suggest that these plant phenols Louis, MO. MNU was a product of K&K Rare and Fine Chemicals. have substantial though variable potential for modifying the risk of skin All other chemicals were obtained in the purest form commercially tumorigenicity induced by a wide variety of chemicals and of these tannic available. acid was shown to have maximal chemoprotective effects. Animals. Six-wk-old female SENCAR mice were obtained from National Cancer Institute-Frederick Cancer Research Facility, Be- thesda, MD. Female BALB/c mice, aged 6 wk, were obtained from INTRODUCTION Charles River Laboratory (Wilmington, MA). The mice were shaved with electric clippers and Nair depilatory was applied 1 day prior to It is now widely accepted that the initiation of carcinogenesis by PAHs3 relates to the metabolic activation of the parent the beginning of the experiment. Only those mice that were not in the hair regrowth cycle were selected for studies. compound into highly reactive ultimate carcinogenic metabo Effect of Plant Phenols in an Initiation and Promotion Protocol. Four lites which eovalently bind to DNA (1-5). This knowledge has hundred SENCAR mice were divided into four groups of 100 animals led to a search for nontoxic inhibitors of these metabolic each and 20 animals from each group were topically treated with a processes. For example there has been growing interest in the single application of acetone, tannic acid, quercetin, myricetin, or identification of naturally occurring dietary factors as potential anthraflavic acid (200 Â«Â¿g/animalin0.2 ml acetone) daily for 7 consec anticarcinogens (6-9). The identification and characterization utive days. At these test concentrations each plant phenol was soluble of such dietary compounds and the definition of their antitumor and the solutions were prepared fresh daily and were applied within 2 h of preparation. Twenty-four h after the last treatment with the plant effects could lead to important strategies for reducing the risk phenols animals from each group received a single topical application of human cancer (10, 11). of either DMBA (40 nmol), BP (400 nmol), MNU (20 *Â¿mol),oracetone In the recent past several naturally occurring plant phenols (control) as the initiating agent. The dosages of PAHs used are standard including tannic acid were shown to inhibit the mutagenicity of for skin tumor induction in SENCAR mice as described earlier (IS) bay-region diol-epoxides of PAHs in the Ames mutagen assay while the MNU dose was chosen based on preliminary experiments using Salmonella typhimurium and in Chinese hamster V79 and as described by Waynforth and Magee (16). Seven days following the single initiating dose of PAHs or MNU each animal then received Received 9/14/87; revised 1/15/88; accepted 1/27/88. The costs of publication of this article were defrayed in part by the payment TPA (3.24 nmol) twice weekly. The experiment was terminated only of page charges. This article must therefore be hereby marked advertisement in after 100% of the animals in each treatment group developed neo accordance with 18 U.S.C. Section 1734 solely to indicate this fact. plasms. Skin tumor formation was recovered weekly and tumors greater 1To whom correspondence should be addressed, at Veterans Administration than 1 mm in diameter were included in the cumulative total only if Medical Center, 10701 East Boulevard, Cleveland, OH 44106. 'Present address: Industrial Toxicology Research Centre, P. O. Box 80, they persisted for 2 wk or more. No skin neoplasms occurred in any Lucknow 226001, India. mice treated with acetone alone or with plant phenol alone. The latent 3The abbreviations used are: PAH, polycyclic aromatic hydrocarbon; BP, periods were computed by the method of Shimkin and Andervont (17) benzo(a)pyrene; BP-7,8-diol, (Â±)-70,8a-dihydroxy-7,8-dihydrobenzo(a)pyrene; as described earlier (18). Statistical significance was determined by a DMBA, 7,12-dimethylbenz(a)anthracene; BPDE-I, (+)-70,8a-dihydroxy-9a,10a- single-tailed Student's r test. epoxy-7,8,9,10-tetrahydrobenzo(a)pyrene or (+)-anti-BPDE; TPA, 12-O-tetradecanoylphorbol-13-acetate; MNU, A'-methyl-A'-nitrosourea; MCA, 3-methyl Effect of Plant Phenols in Complete Carcinogenesis Protocol. Two cholanthrene. hundred BALB/c mice were divided into five groups of 40 each. 2361

Animals in group 1 received skin applications of 0.2 ml of acetone daily for seven days whereas the mice of the remaining group received daily skin applications of 3.0 ^mol per mouse of either tannic acid, quercetin, NONE anthraflavic acid, or 0.75 Â¿tinolper mouse of myricetin, in 0.2 ml of TANNIC ACID â€¢QUERCETIN acetone. After this pretreatment regimen 20 animals from each group Â° MYRICETIN received twice-weekly applications of the plant phenol followed 30 min * ANTHRAFLAVIC ACID later by 1.5 Â¿Â¿molofMCA in 0.2 ml of acetone whereas the remaining mice of each group received only plant phenol in acetone. This treat ment was repeated in each group for 16 wk at which time the experiment was terminated. Skin tumor formation was recorded weekly, and tumors greater than 1 mm in diameter were included in the cumulative total only if they persisted for 2 wk or more. No skin neoplasm occurred in any mice treated with acetone alone or plant phenols alone.

RESULTS Effect of Topical Applications of Plant Phenols on DMBA- induced Skin Tumorigenesis. The effect of pretreatment with 12 plant phenols on skin tumor induction in DMBA-initiated and TPA-promoted SENCAR mice is shown in Fig. 1. The animals pretreated with each plant phenol showed considerable prolon 100 - gation of the latent period prior to the onset of tumor devel opment. The first tumor appeared after 6, 5, 5, and 4 wk of OC treatment with DMBA in the tannic acid-, quercetin-, myrice o tin-, and anthraflavic acid-pretreated animals, respectively, as 80 - compared to 3 wk in the corresponding control group without plant phenol pretreatment. At the termination of the experi ment after 12 wk the cumulative number of tumors in 20 mice â€¢o- in the non-plant phenol-pretreated group was 680, whereas LU only 351, 368, 302, and 409 tumors were recorded in the groups O pretreated with tannic acid, quercetin, myricetin, and anthra 40 - flavic acid, respectively (Fig. 1/1). After 6 wk of topical appli cation of DMBA only 30, 55, 25, and 55% of the mice in the tannic acid-, quercetin-, myricetin-, and anthraflavic acid-pre ut treated groups demonstrated any neoplasm as compared to O 100% of the animals in the control group without plant phenol OC pretreatment (Fig. IB). The data in Table 1 show skin tumors per mouse as a function of the number of weeks on test. At each time point highly significant protection against skin tumor WEEKS ON TEST formation was evident in animals pretreated with each plant Fig. 1. Effect of tannic acid, quercetin, myricetin, and anthraflavic acid on phenol. By 12 wk of testing, the number of tumors per mouse DMBA-induced skin tumorigenesis in SENCAR mice. Female (6-wk-old) SEN- in the non-plant phenol-pretreated group was 34.00, whereas CAR mice were treated with different plant phenols as described in "Materials and Methods." Twenty-four h after the last application of the plant phenol the corresponding numbers in the groups pretreated with tannic animals received topical application of a single initiating dose of DMBA. One wk acid, quercetin, myricetin, and anthraflavic acid were 17.55, later tumors were promoted by twice-weekly application of TPA. The cumulative 18.40, 15.10, and 20.45, respectively. number of tumors (.-I) and the percentage of mice with tumors (II) were plotted Effect of Topical Applications of Plant Phenols on BP-induced as a function of the number of weeks on test. Skin Tumorigenesis. The data shown in Fig. 2 represent the effect of pretreatment with plant phenols on the cumulative 100% of the animals in control group without plant phenol number of tumors and percentage of mice with tumors in BP- pretreatment (Fig. IB). The data in Table 1 represent skin initiated and TPA-promoted SENCAR mice. The animals pre tumors per mouse as a function of the number of weeks on test. treated with each plant phenol developed significantly fewer At each time point highly significant protection against BP- skin tumors and the latent period prior to the onset of tumor induced skin tumor formation was evident in animals pretreated development was also considerably prolonged in these animals. with each plant phenol. By 12 wk of testing, the number of The first tumor appeared after 9, 8, 8, and 7 wk of treatment tumors per mouse in the non-plant phenol-pretreated group with BP in the tannic acid-, quercetin-, myricetin-, and anthra was 10.90, whereas the corresponding numbers in the groups flavic acid-pretreated animals, respectively, as compared to 5 pretreated with tannic acid, quercetin, myricetin, and anthra wk in the corresponding control group without plant phenol flavic acid were 2.90, 4.75, 5.25, and 5.60, respectively. pretreatment. At the termination of the experiment after 14 wk Effect of Topical Applications of Plant Phenols on MNU- the cumulative number of tumors in 20 mice in the non-plant induced Skin Tumorigenesis. Fig. 3 shows the effect of pretreat phenol-pretreated group was 243, whereas only 81, 124, 127, ment with plant phenols on cumulative number of tumors and and 136 tumors were registered in the tannic acid-, quercetin-, percentage of mice with tumors in MNU-initiated and TPA- myricetin-, and anthraflavic acid-pretreated groups, respectively promoted SENCAR mice. The animals pretreated with each (Fig. 2A). After 10 wk of test 40, 65, 70, and 95% of the mice plant phenol developed significantly fewer skin tumors and the in the tannic acid-, quercetin-, myricetin-, and anthraflavic acid- latent period prior to the onset of tumor development was also pretreated groups demonstrated neoplasms as compared to considerably prolonged in these animals. The first tumor ap- 2362

Table 1 Effect of topical application of plant phenols on DMBA-, BP-, and 280 - MNV-induced skin tumorigenesis in an initiation and promotion protocol in SENCAR mice en â€¢NONE Female SENCAR mice were maintained on food and water ad libitum. Plant OC 240 o TANNIC ACID phenols were topically applied as described in "Materials and Methods." Twenty- O â€¢QUERCEâ„¢ four h after the last plant phenol application animals received DMBA, BP, or o MYRICETIN MNU as the initiating agent. For other details see text. Data are the mean Â±SE * ANTHRAFLAVIC of 20 animals in each group. ACID

O 160 PretreatmentExperiment usedDMBADMBADMBADMBADMBABPBPBPBPBPMNUMNUMNUMNUMNU16wk12.25wk32.10 wk34.00 O 120 - 1SolventTannic Â±2.101.50 Â±3.1811. Â±2.7317.55 LU acidQuercetinMyricetinAnthraflavicÂ±0.55"1.90 50Â±2.38Â°14.20 Â±1.45Â°18.40Â± Â±0.49Â°0.85 Â±2.40Â°10.95 1.45Â°15.10Â± 80 - Â±0.35Â°4.05 Â±2.16Â°16.95 2.04Â°20.45 < acidExperiment Â±1.17Â°0.70 Â±2.60Â°3.70 Â±1.99Â°10.90Â± Z 2SolventTannic Â±0.18NTF*NTFNTFNTFNTFNTFNTFNTFNTFTumors/mouse9Â±0.550.35 1.072.90 O acidQuercetinMyricetinAnthraflavic Â±0.15Â°1.20 Â±0.67Â°6.05 Â±0.39Â°0.65 Â±0.90Â°5.25 Â±0.24Â°1.20 Â±1.07Â°5.60 acidExperiment Â±0.36Â°2.00 Â±0.73Â°4.80

3SolventTannic V) ' Â±0.530.35 Â±1.16Â°1.00 acidQuercetinMyricetinAnthraflavic Â±0.13Â°0.60 Â±0.32Â°1.55 OC Â±0.18Â°0.65 Â±0.35Â°2.30 o Â±0.20Â°0.65 Â±0.56Â°2.25 3 so acidCarcinoger Â±0.20Â°12 Â±0.46Â° Â°Statistically significant from the control (/' < 0.01) (Student's t test). * NTF, no tumor found. peared after 9, 8, 8, and 7 wk of treatment with MNU in the u tannic acid-, quercetin-, myricetin-, and anthraflavic acid-pre- u treated animals, respectively, as compared to 7 wk in the i corresponding control group without plant phenol pretreat ment. At the termination of the experiment after 17 wk the cumulative number of tumors in 20 mice in the non-plant LJJ phenol-pretreated group was 155, whereas only 53, 61, 89, and Ote. 91 tumors were recorded in the tannic acid-, quercetin-, myri W cetin-, and anthraflavic acid-pretreated groups, respectively a â€”râ€” (Fig. 3/4). The data in Table 1 also show skin tumors per mouse io 12 as a function of the number of weeks on test. At each time WEEKS ON TEST point highly significant protection against MNU-induced skin Fig. 2. Effect of tannic acid, quercetin, myricetin, and anthraflavic acid on Bl> indiimi skin tumorigenesis in SENCAR mice. The treatment schedule was tumor formation was evident in animals pretreated with each identical to that described in Fig. 1 with the exception that BP was used as a plant phenol. By 12 wk of testing, the number of tumors per tumorigen. The cumulative number of tumors (.1) and the percentage of mice mouse in the non-plant phenol-pretreated group was 4.80, with tumors (It) were plotted as a function of the number of weeks on test. whereas corresponding numbers in the groups pretreated with tannic acid, quercetin, myricetin, and anthraflavic acid were numbers of weeks on test. At each time point statistically 1.00, 1.55, 2.30, and 2.25, respectively. Effect of Topical Applications of Plant Phenols on MCA- significant protection against skin tumor formation was evident in animals treated with tannic acid, quercetin, and myricetin induced Skin Tumorigenesis. The tumor induction studies were whereas the protective effects were only modest with anthra conducted in groups of BALB/c mice receiving MCA simulta flavic acid. At the termination of the experiment at 16 wk the neously with or without the topical application of plant phenols tumors per mouse were found to be inhibited 62, 57, 47, and in a complete carcinogenesis protocol and the results are shown 28% by tannic acid, quercetin, myricetin, and anthraflavic acid, in Fig. 4. Tumor data are presented as cumulative number of tumors (Fig. 4/4) and the percentage of mice with tumors (Fig. respectively. 4B) as a function of weeks on test. In each case each of the plant phenols was found to afford significant protection against DISCUSSION skin tumor induction. For example, it can be seen that the first skin tumor appeared 8 wk after MCA administration in animals The naturally occurring plant phenols tannic acid, quercetin, receiving MCA alone, but appeared after 10, 10, 10, and 9 wk myricetin, and anthraflavic acid are widely distributed in the of treatment with the carcinogen in the tannic acid-, querce plant kingdom (19-22) and many of these compounds are tin, myricetin-, and anthraflavic acid-treated animals, respec ingested in the human diet. It has been estimated that some tively. At the termination of the experiment after 16 wk the individuals ingest as much as l g of the plant phenols per day cumulative number of tumors in 20 mice without plant phenol in their diet (23). The data in this study demonstrate that the pretreatment was 232, whereas only 99, 100, 123, and 166 topical application of tannic acid, quercetin, myricetin, and tumors were recorded in the tannic acid-, quercetin-, myrice anthraflavic acid affords significant protection against BP-, tin-, and anthraflavic acid-treated groups, respectively. The data DMBA-, MNU-, and MCA-induced skin tumorigenesis. In in Table 2 show skin tumors per mouse as a function of the general the order of anticarcinogenic potency of these plant 2363 Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1988 American Association for Cancer Research. PROTECTION OF SKIN TUMORIGENESIS BY PLANT PHENOLS

â€¢NONE <0 o TANNIC ACÂ» o 27Â° â€¢NONE â€¢QUERCETIN o TANNIC ACID = 240 â€¢QUERCETIN o MYRICETIN 210 Li. â€¢ANTHRAFLAVIC ACID O 180

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t 4 â€¢ â€¢ 10 li 14 1Â« It WEEKS ON TEST Fig. 3. Effect of tannic acid, quercetin, myricetin, and anthraflavic acid on U! 20 - MNU-induced skin tumorigenesis in SENCAR mice. The treatment schedule was O identical to that described in Fig. 1 except MNU was used as the initiating agent. OC The cumulative number of tumors (A) and the percentage of mice with tumors LU (B) were plotted as a function of the number of weeks on test. O. o 10 11 12 14 15 10 phenols was found to be tannic acid > myricetin > quercetin > WEEKS ON TEST anthraflavic acid. Fig. 4. Effect of tannic acid, quercetin, myricetin, and anthraflavic acid on Phenolic plant constituents have been shown to possess sev MCA-induced skin tumorigenesis in BALB/c mice. Female (6-wk-old) BALB/c eral activities which may influence the tumorigenic effect of mice were treated with different plant phenols as described in "Materials and certain chemical carcinogens. Some plant phenols have been Methods." The cumulative number of tumors (.â€¢()andthe percentage of mice shown to inhibit the activity of cytochrome P-450-dependent with tumors (B) were plotted as a function of the number of weeks on test. enzymes that metabolize drugs and carcinogens (24, 25). For Table 2 Effect of topical application of plant phenols in a complete skin example our studies (13) indicated that topical applications of carcinogenesis protocol using MCA in BALB/c mice tannic acid, quercetin, myricetin, and anthraflavic acid to SEN- Female Balb/c mice were maintained on food and water ad libitum. Plant CAR mice inhibit epidermal PAH metabolism. Some of the phenols were topically applied as described in "Materials and Methods." For plant phenols have been shown to possess antioxidant activity other details see text. Data are the mean Â±SE of 20 animals in each group. and inhibit the formation of nitrosamines (26, 27). Our recent studies have also shown that tannic acid, quercetin, myricetin, PretreatmentSolvent wk1.15 wk11.61 and anthraflavic acid inhibit the binding of [3H]BP, [3H]BP- Â±0.05 Â±0.28 Â±0.57 7,8-diol, and [3H]DMBA to epidermal DNA in SENCAR mice Tannic acid NTF" 0.55 Â±0.20* 4.47 Â±0.39* Quercetin NTF 0.55 Â±0.18* 5.00 Â±0.32* 6.15 Â±0.32' (14). In these studies it was shown that tannic acid was the Myricetin NTF 0.40 Â±0.20* most potent inhibitor of BPDE-I-deoxyguanosine adduct for Anthraflavic acid8wk0.05 NTFTumors/mouse121.25 Â±0.1016 8.30 Â±0.40" mation in epidermis and lungs of SENCAR mice (14). " NTF, no tumor found. * Statistically significant from the control (/' < 0.01) (Student's t test). Very few studies have assessed the ability of these plant phenols to protect against the development of tumors in exper imental animals. Chang et al. (28) have shown that myricetin, caffeic acid have been reported to inhibit the formation of BP- quercetin, and ellagic acid were moderately effective inhibitors induced forestomach tumors in the mouse (31). Recent studies of the skin tumorigenic activity of BPDE-I but could not have indicated that quercetin inhibits arachidonate-5-lipoxy- demonstrate statistically significant effects on the tumorigenic- genase activity in vitro (32) and the tumor-promoting activity ity of BP in CD-I mouse skin. Prior studies have shown that of TPA on mouse skin (33). Tannic acid, quercetin, myricetin, quercetin inhibits the tumorigenicity of BP in mouse skin (29, and anthraflavic acid have been shown to be highly effective 30) while some other plant phenols including ferrulic acid and inhibitors of the mutagenicity of BP-7,8-diol-9,10-epoxide-2 (8, 2364 Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1988 American Association for Cancer Research. PROTECTION OF SKIN TUMORIGENESIS BY PLANT PHENOLS

9) while several other flavonoids are much less effective in this of bay-region diol-epoxides of polycyclic aromatic hydrocarbons by tannic acid, hydroxylated anthraquinones and hydroxylated cinnamic acid deriva regard. Huang et al. (8,9) have proposed that the an timu tagen ic tives. Carcinogenesis (Lond.), 6:237-242, 1985. activity of these compounds is due to the disappearance of bay- 10. Workshop Conference on Nutrition in Cancer Causation and Prevention. region diol-epoxides of PAHs as a result of their interaction Cancer Res., 43: 2385s-2519s, 1983. 11. Correa, P. EpidemiolÃ³gica! correlations between diet and cancer frequency. with the reactive metabolites. This may explain the antitu- Cancer Res., 41: 3685-3689, 1981. 12. Wood, A. W., Huang, M. T., Chang, R. L., Newmark, H. L., Lehr, R. E., morigenic activity of these plant phenols as observed in this Yagi, H. Sayer, J. M., Jerina, D. M., and Conney, A. H. Inhibition of the study. mutagenicity of bay-region diol epoxides of polycyclic aromatic hydrocarbons by naturally occurring plant phenols: exceptional activity of ellagic acid. If the protective effect of these plant phenols against skin Proc. Nati. Acad. Sci. USA, 79: 5513-5517, 1982. tumorigenicity by PAHs is entirely related to their inhibitory 13. Das, M., Mukhtar, H., Bik, D. P., and Bickers, D. R. Inhibition of epidermal effect on metabolism of carcinogens and their subsequent bind xenobiotic metabolism in SENCAR mice by naturally occurring plant phen ing to DNA, then these plant phenols should have no antitu- ols. Cancer Res., 47: 760-766, 1987. 14. Das, M., Khan, W. A., Asokan, P., Bickers, D. R., and Mukhtar, H. Inhibition morigenic effect against skin tumorigenicity by chemicals like of polycyclic aromatic hydrocarbon-DNA adduci formation in epidermis and lungs of SENCAR mice by naturally occurring plant phenols. Cancer Res., MNU which do not require metabolism to elicit tumorigenicity. 47:167-113, 1987. However, our results indicate that all of the plant phenols tested 15. Mukhtar, H., Das, M., and Bickers, D. R. Skin tumor initiating activity of showed significant protection against MNU-induced skin tu therapeutic crude coal tar as compared to other polycyclic aromatic hydro morigenicity. MNU is a direct-acting carcinogen whose cancer- carbons in SENCAR mice. Cancer Lett., 31:147-151, 1986. 16. Waynforth, H. B., and Magee, P. N. The effect of various doses and schedules causing effect is related to alkylation of DNA and single strand of administration of W-methylWV-nitrosourea, with and without croton oil breaks (34-36). Thus, the possible mechanism of protective promotion on skin papilloma production in BALB/c mice. Gann, / 7: 439- 448, 1975. effect of these plant phenols may be due to an inhibitory effect 17. Shimkin, M. B., and Andervont, H. B. Comparative carcinogenicity of three on the binding of the ultimate carcinogen to target tissue DNA. carcinogenic hydrocarbons. J. Nati. Cancer Inst., A-57-62, 1940. 18. Mukhtar, H., DelTito, B. J., Jr., Das, M., Charniack, E. P., Charniack, A. Teel (37) has shown that the anticarcinogenic and antimuta D., and Bickers, D. R. Clotrimazole, an inhibitor of epidermal genie activity of plant phenols, for example, ellagic acid, is due benzo(a)pyrene metabolism and DNA binding and carcinogenicity of the to an interaction of the compound with target tissue DNA hydrocarbon. Cancer Res., 44:4233-4240, 1984. 19. Harborne, J. B., Mabry, T. J., and Mabry, H. The Flavonoid, Vols. 1 and 2. which in turn blocks the site(s) of DNA to electrophilic attack New York: Academic Press, 1975. by reactive carcinogenic moieties. The studies of Dixit and Gold 20. Bete-Smith, E. C. Detection and determination of ellagitannins. Phytochem- istry, 11:1153-1156, 1972. (38) have shown that ellagic acid can inhibit the activity of the 21. Haslam, E. Vegetable tannins. Recent Adv. Phytochem., 12:475-523,1979. direct-acting mutagen MNU in S. typhimurium TA 100. Fur 22. Fairbairn, J. W. The Pharmacology of Plant Phenolics. New York: Academic thermore, these workers have reported that the antimutagenic Press, 1959. 23. Brown, J. P. A review of the genetic effects of naturally occurring flavonoids, activity of ellagic acid against MNU is due to inhibition in anthraquinones and related compounds. MutÃ¢t.Res., 75: 243-277, 1980. methylation at the O6 position of guanine in double stranded 24. Buening, M. K., Chang, R. L., Huang, M. T., Fortner, J. G., Wood, A. W., and Conney, A. H. Activation and inhibition of benzo(a)pyrene and aflatoxin DNA (38). H, metabolism in human liver microsomes by naturally occurring flavonoids. In summary, our results indicate that plant phenols, partic Cancer Res., 41:67-72, 1981. 25. Lasker, J. M., Huang, M. T., and Conney, A. H. In vitro and in vivoactivation ularly tannic acid, have antitumorigenic activity against PAHs of oxidative drug metabolism by flavonoids. J. Pharmacol. Exp. I her., 229: and MNU when applied in vivo and one possible mechanism 162-170, 1984. may be an alteration in target tissue macromolecular binding 26. Kuenzig, W., Chau, J., Norkus, E., Holowaschenko, H., Newmark, H., Mergens, W., and Conney, A. H. Caffeic acid and femilic acid as blockers of reactive carcinogenic species. of nitrosamine formation. Carcinogenesis (Lond.), 5: 309-313, 1984. 27. Pignatelli, B., Bereziat, J. C., Descotes, G., and Bartsch, H. Catalysis of nitrosamine in vitro and in vivo rats in by catechin and resorcinol and ACKNOWLEDGMENTS inhibition by chlorogenic acid. Carcinogenesis (Lond.), I: 1045-1049, 1982. 28. Chang, R. L., Huang, M. T., Wood, A. W., Wong, C. Q., Newmark, H. L., Supported in part by NIH Grants ES-1900, CA-38028, and AM Yagi, H., Sayer, J. M., Jerina, D. M., and Conney, A. H. 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Downloaded from cancerres.aacrjournals.org on September 29, 2021. © 1988 American Association for Cancer Research. Exceptional Activity of Tannic Acid among Naturally Occurring Plant Phenols in Protecting against 7,12-Dimethylbenz( a )anthracene-, Benzo(a)pyrene-, 3-Methylcholanthrene-, and N -Methyl-N-nitrosourea-induced Skin Tumorigenesis in Mice

Hasan Mukhtar, Mukul Das, Wasiuddin A. Khan, et al.

Cancer Res 1988;48:2361-2365.

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