Inhibitory Effects of Luteolin Isolated from Ixeris sonchifolia Hance on the Proliferation of HepG2 Human Hepatocellular Carcinoma Cells

  • Yee, Su-Bog (Department of Pharmacy, Pusan National University) ;
  • Lee, Jung-Hwa (Department of Pharmacy, Pusan National University) ;
  • Chung, Hae-Young (Department of Pharmacy, Pusan National University) ;
  • Im, Kwang-Sik (Department of Pharmacy, Pusan National University) ;
  • Bae, Song-Ja (Department of Food and Nutrition, Silla University) ;
  • Choi, Jae-Soo (Faculty of Food Science and Biotechnology, Pukyong National University) ;
  • Kim, Nam-Deuk (Department of Pharmacy, Pusan National University)
  • Published : 2003.02.01

Abstract

We investigated the anti-proliferative effects of luteolin and apigenin, isolated from Ixeris sonchifolia Hance, on HepG2 human hepatocellular carcinoma cells. In MTT assay luteolin showed more efficient anti-proliferative effects on cells than apigenin did. According to propidium iodide staining and flow cytometry studies, we postulated that these effects might be a result of cell cycle arrest. Hence we examined the changes of protein expressions related to cell cycle arrest. Western blotting data demonstrated that the down-regulated expression of CDK4 was correlated to the increase of p53 and CDK inhibitor $p21^{WAF1/CIP1}$ protein. These data suggest that luteolin may have potential as an anti-cancer agent.

Keywords

References

  1. Bae, S. J., Kim, N. H., Koh, J. B., Roh, S. B., and Jung, B. M., Effects of godulbaegi (lxeris sonchifolia H.) diets on enzyme activities of CC$I_{4}$ induced hepatotoxicity in rats. J. Korean Nutr. Soc., 30, 19-24 (1997a)
  2. Bae, S. J., Kim, N. H., Ha, B. J., Jung, B. M., and Roh, S. B., Effects of godulbaegi leaf extracts on CC$I_{4}$ -induced hepatotoxicity in rats. J. Korean Soc. Food Sci. Nutr., 26, 137-143 (1997b)
  3. Bae, S. J., Kim, N. H., Roh, S. B., and Jung, B. M., The effects of godulbaegi extracts on the fluidity of phospholipid liposomes by DSC. J. Korean Soc. Food Sci. Nutr., 27, 518-524 (1998a)
  4. Bae, S. J., Roh, S. B., and Jung, B. M., Effects of godulbaegi extracts on the stability and fluidity of phospholipid liposomal membranes. J. Korean Soc. Food Sci. Nutr., 27, 508-517 (1998b)
  5. Casagrande F. and Darbon J. M., Effects of structurally related flavonoids on cell cycle progression of human melanoma cells: regulation of cyclin-dependent kinases CDK2 and CDK1. Biochemical Pharmacology, 61, 1205-1215 (2000) https://doi.org/10.1016/S0006-2952(01)00583-4
  6. Chiu, H. F., Lin, C. C., Yang, C. C., and Yang, R., The pharmacological and pathological studies, on several hepatic protective crude drugs from Taiwan (II). Am. J. Chin. Med., 17, 17-30 (1989) https://doi.org/10.1142/S0192415X89000048
  7. Chung, H. S., Cytotoxic triterpenoids on human cancer cell lines from Ixeris sonchifolia. Food Sci. Biotechnol., 9, 364-367 (2000)
  8. Chung, H. S., Guaianolide sesquiterpene lactone from Ixeris sonchifolia Hance with cytotoxicity in cultured human stomach and colon cancer cell lines. Food Sci. Biotechnol., 10, 433-436 (2001)
  9. El-Deiry, W. S., Tokino, T., Velculescu, V. E., Levy, D. B., Parsons, R., Trent, J. M., Lin, D., Mercer, W. E., Kinzler, K. W., and Vogelstein, B., WAF1, a potential mediator of p53 tumor suppression. Cell, 75, 817-825 (1993) https://doi.org/10.1016/0092-8674(93)90500-P
  10. Elledge, S. J., Cell cycle checkpoints: preventing an identity crisis. Science, 274, 1664-1672 (1996) https://doi.org/10.1126/science.274.5293.1664
  11. Gupta, S., Afaq, F., and Mukhtar, H., Selective growth-inhibitory, cell-cycle deregulatory and apoptotic response of apigenin in normal versus human prostate carcinoma cells. Biochem. Biophys. Res. Cammun., 287, 914-920 (2001) https://doi.org/10.1006/bbrc.2001.5672
  12. Hartwell, L. H. and Kastan, M. B., Cell cycle control and cancer. Science, 266, 1821-1828 (1994) https://doi.org/10.1126/science.7997877
  13. Hirano, T., Oka, K., and Akiba, M., Antiproliferative effects of synthetic and naturally occurring flavonoids on tumor cells of the human breast carcinoma cell line, ZR-75-1. Res. Commun. Chem. Pathol. Pharmacol., 64, 69-78 (1989)
  14. Im, E. O., Choi, Y. H., Paik, K. J., Suh, H., Jin, Y., Kim, K. W., Yoo, Y. H., and Kim, N. D., Novel bile acid derivatives induce apoptosis via a p53-independent pathway in human breast carcinoma cells. Cancer Lett., 163, 83-93 (2001) https://doi.org/10.1016/S0304-3835(00)00671-6
  15. Knowles, L. M., Zigross, D. A., Tauber, R. A., Hightower, C., and Milne, J. A., Flavonoids suppress androgen-independent human prostate tumor proliferation. Nutr. Cancer, 38, 116-122 (2000) https://doi.org/10.1207/S15327914NC381_16
  16. Lin, S. C., Lin, C. C., Lin, Y. H., and Yao, C. J., Hepatoprotective effects of Taiwan folk medicine: Ixeris chinensis (Thunb.) Nak. on experimental liver injuries. Am. J. Chin. Med., 22, 243-254 (1994) https://doi.org/10.1142/S0192415X94000309
  17. Li, Y. C., Hung, C. F., Yeh, F. T., Lin, J. P., and Chung, J. G., Luteolin-inhibited arylamine N-acetyltransferase activity and DNA-2-aminofluorene adduct in human and mouse leukemia cells. Food Chem. Toxicol., 39, 641-647 (2001) https://doi.org/10.1016/S0278-6915(01)00009-6
  18. Lu, K. L., Chang, Y. S., Ho, L. K., Lin, C. C., and Tsai, C. C., The evaluation of the therapeutic effect of tao-shang-tsao on alpha-naphthylisothiocyanate and carbon tetrachloride-induced acute liver damage in rats. Am. J. Chin. Med., 28, 361-370 (2000) https://doi.org/10.1142/S0192415X00000428
  19. Morgan, D. O., Principles of CDK regulation. Nature, 374, 131-134 (1995) https://doi.org/10.1038/374131a0
  20. Park, S. S., Studies on the constituents with biological activities of Ixeris sonchifolia Hance. Korean Biochem. J., 10, 241-252 (1977)
  21. Pettit, G. R., Hoard, M. S., Doubek, D. L., Schmidt, J. M., Pettit, R. K., Tackett, L. P., and Chapuis, J. C., Antineoplastic agents 338. The cancer cell growth inhibitory. Constistituents of Terminalia arjuna (Combretaceae). J. Ethnopharmacol., 53, 57-63 (1996) https://doi.org/10.1016/S0378-8741(96)01421-3
  22. Pike, B. R., Zhao, X., Newcomb, J. K., Wang, K. K., Posmantur, R. M., and Hayes, R. L., Temporal relationships between de novo protein synthesis, calpain and caspase 3-like protease activation, and DNA fragmentation during apoptosis in septohippocampal cultures. J. Neurosci. Res., 52, 505-520 (1998) https://doi.org/10.1002/(SICI)1097-4547(19980601)52:5<505::AID-JNR3>3.0.CO;2-G
  23. Post, J. F. and Varma, R. S., Growth inhibitory effects of bioflavonoids and related compounds on human leukemic CEMC1 and CEM-C7 cells. Cancer Lett., 24, 207-213 (1992)
  24. Sherr, C. J. and Roberts, J. M., Inhibitors of mammalian G1 cyclin-dependent kinases. Genes Dev., 9, 1149-1163 (1995) https://doi.org/10.1101/gad.9.10.1149
  25. Sherr, C. J., Cancer cell cycles. Science, 274, 1672-1677 (1996) https://doi.org/10.1126/science.274.5293.1672
  26. Shimoi, K., Saka, N., Kaji, K., Nazawa, R., and Kinae, N., Metabolic fate of luteolin and its functional activity at focal site. Biofactors, 12, 181-186 (2000) https://doi.org/10.1002/biof.5520120129
  27. Shin, S. C., Studies on the chemical components of wild Korean lettuce (Youngia sonchifolia Max.). J. Korean Agric. Chem. Soc., 31, 134-137 (1988)
  28. Shin, S. C., Exploitation of the biologically active components in Youngia sonchifolia Max. J. Korean Agric. Chem. Soc., 36, 261-266 (1993)
  29. Suh, J., Jo, Y., Kim, N. D., Bae, S. J., and Im, K. S. (2002) Cytotoxic constituents of the leaves of Ixeris sonchifolia. Arch. Pharm. Res., 25, 289-292 (2002) https://doi.org/10.1007/BF02976628
  30. Tada, H., Shiho, O., Kuroshima, K., Koyama, M., and Tsukamoto, K., An improved colorimetric assay for interleukin 2. J. Immunol. Methods, 93, 157-165 (1986) https://doi.org/10.1016/0022-1759(86)90183-3
  31. Wang, I. K., Lin-Shiau, S. Y., and Lin, J. K., Induction of apoptosis by apigenin and related flavonoids through cytochrome c release and activation of caspase-9 and caspase-3 in leukemia HL-60 cells. Eur. J. Cancer, 35, 1517-1525 (1999) https://doi.org/10.1016/S0959-8049(99)00168-9
  32. Wang, W., Heideman, L., Chung, C. S., Pelling J. C., Koehler K. J., and Birt, D. F., Cell-cycle arrest at G2/M and growth inhibition by apigenin in human colon carcinoma cell lines. Mol. Carcinog., 28, 102-110 (2000) https://doi.org/10.1002/1098-2744(200006)28:2<102::AID-MC6>3.0.CO;2-2
  33. Yin, F., Giuliano, A. E., and Van Herle, A. J., Growth inhibitory effects of flavonoids in human thyroid cancer cell lines. Thyroid, 9, 369-376 (1999a) https://doi.org/10.1089/thy.1999.9.369
  34. Yin, F., Giuliano, A. E., and Van Herle, A, J., Signal pathways involved in apigenin inhibition of growth and induction of apoptosis of human anaplastic thyroid cancer cells (ARO). Anticancer Res., 19, 4297-4303 (1999b)
  35. Yin, F., Giuliano, A. E., Law, R. E., and Van Herle, A. J., Apigenin inhibits growth and induces G2/M arrest by modulating cyclin-CDK regulators and ERK MAP kinase activation in breast carcinoma cells. Anticancer Res., 21, 413-420 (2001)
  36. Young, H. S., Choi, J. S., and Lee, J. H., Further study on the anti-hypercholesterolemic effect of Ixeris sonchifolia. Korean J. Pharmacog., 23, 73-76 (1992a)
  37. Young, H. S., Im, K. S., and Choi, J. S., The pharmaco-chemical study on the plant of Ixeris spp. 2. Flavonoids and free amino acid composition of Ixeris sonchifolia. J. Korean Soc. Food Nutr., 21, 296-301 (1992b)
  38. Young, H. S., Seo, S. S., Lee, K. H., Lee, J. H., and Choi, J. S., The pharmacochemical study on the plant of Ixeris spp. 1. Anti-hypercholesterolemic effect of Ixeris sonchifolia. J. Korean Soc. Food Nutr., 21, 291-295 (1992c)