The centella asiatica juice effects on DNA damage, apoptosis and gene expression in hepatocellular carcinoma (HCC)

BMC Complementary and Alternative Medicine

Abdah Akim¹  Fauziah Othman¹  Asmah Rahmat¹  Sima Ataollahi Eshkoor²  Faridah Hussin¹ 

[1] Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia;Institute of Gerontology, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia

关键词: Hepatocellular carcinoma (HCC);    Gene expression;    Apoptosis;    DNA damage;    Centella asiatica juice;   

Others  :  1220288
DOI  :  10.1186/1472-6882-14-32

 received in 2013-06-27, accepted in 2013-12-09,  发布年份 2014

【 摘 要 】

Background

This paper is to investigate the effects of Centella asiatica on HepG2 (human hepatocellular liver carcinoma cell line). Centella asiatica is native to the Southeast Asia that is used as a traditional medicine. This study aims to determine the chemopreventive effects of the Centella asiatica juice on human HepG2 cell line.

Methods

Different methods including flow cytometry, comet assay and reverse transcription-polymerase chain reaction (RT-PCR) were used to show the effects of juice exposure on the level of DNA damage and the reduction of cancerous cells. MTT assay is a colorimetric method applied to measure the toxic effects of juice on cells.

Results

The Centella asiatica juice was not toxic to normal cells. It showed cytotoxic effects on tumor cells in a dose dependent manner. Apoptosis in cells was started after being exposed for 72 hr of dose dependent. It was found that the higher percentage of apoptotic cell death and DNA damage was at the concentration above 0.1%. In addition, the juice exposure caused the reduction of c-myc gene expression and the enhancement of c-fos and c-erbB2 gene expressions in tumor cells.

Conclusions

It was concluded that the Centella asiatica juice reduced liver tumor cells. Thus, it has the potential to be used as a chemopreventive agent to prevent and treat liver cancer.

【 授权许可】

   
2014 Hussin et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 5.

Figure 6.

Figure 7.

【 参考文献 】

• [1]Parikh S, Hyman D: Hepatocellular cancer: a guide for the internist. Am J Med 2007, 120(3):194-202.
• [2]Olsen SK, Brown RS, Siegel AB: Review: hepatocellular carcinoma: review of current treatment with a focus on targeted molecular therapies. Therap Adv Gastroenterol 2010, 3(1):55-66.
• [3]Oliveria SA: The role of epidemiology in cancer prevention. Soc Exp Biol Med 1997, 216(2):142-150.
• [4]Firenzuoli F, Gori L: Herbal medicine today: clinical and research issues. eCAM 2007, 4(Suppl 1):37-40.
• [5]Yoo HH, Park JH, Kwon SW: In vitro cytotoxic activity of some Korean medicinal plants on human cancer cell lines: enhancement in cytotoxicity by heat processing. Phytother Res 2007, 21(9):900-903.
• [6]Rosalizan M, Rohani M, Khatijah I, Shukri M: Physical characteristics, nutrient contents and triterpene compounds of ratoon crops of Centella asiatica at three different stages of maturity. J Trop Agric Food Sci 2008, 36(1):43-51.
• [7]Mahanom H, Azizah A, Suhaila M, Nazamid S, Maznah I, Hair BM: Effect of Centella asiatica extract and powder on hydrogen peroxide-treated Sprague-Dawley rats. J Trop Agric Food Sci 2011, 39(1):000-000.
• [8]Pittella F, Dutra RC, Junior DD, Lopes MT, Barbosa NR: Antioxidant and cytotoxic activities of Centella asiatica (L) Urb. Int J Mol Sci 2009, 10(9):3713-3721.
• [9]Coldren CD, Hashim P, Ali JM, Oh S-K, Sinskey AJ, Rha C: Gene expression changes in the human fibroblast induced by Centella asiatica triterpenoids. Planta Med 2003, 69(8):725-732.
• [10]Amati B, Frank SR, Donjerkovic D, Taubert S: Function of the c-myc oncoprotein in chromatin remodeling and transcription. Biochim Biophys Acta 2001, 1471(3):135-145.
• [11]Jonas AN, Kirsteen HM, Ulrich BK, Helene P, Troy AB, Cleveland JL: Mnt loss triggers c-myc transcription targets proliferation on apoptosis and transformation. Mol Cell Biol 2004, 24(4):1560-1569.
• [12]Tesarik J, Garrigosa L, Mendoza C: Estradiol modulates breast cancer cell apoptosis: a novel nongenomic steroid action relevant to carcinogenesis. Steroids 1999, 64(1-2):22-27.
• [13]Scott GK, Dodson JM, Montgomery PA, Johnson RM, Sartup JC: p185HER2 signal transduction in breast cancer cells. J Biol Chem 1991, 266(22):14300-14305.
• [14]Jiang Y, Zhang Y, Luan J, Duan H, Zhang F, Yagasaki K, Zhang G: Effects of bufalin on the proliferation of human lung cancer cells and its molecular mechanisms of action. Cytotechnology 2010, 62(6):573-583.
• [15]McKelvey-Martin VJ, Green MHL, Schmezer P, de Mjo BMP, Collinns M: The single cell gel electro (comet assay): a European review. Mutat Res 1993, 288(1):47-63.
• [16]Eshkoor SA, Ismail P, Abd. Rahman S, Moin S, Adon MY: The association of DNA damage level with early age at the occupational exposure in the mechanical workshops workers. Asian J Biotechnol 2012, 4(2):83-91.
• [17]Coucouvanis EC, Jones PP: Changes in protooncogene expression correlated with general and sex specific differentiation in murino primordial germ cells. Mech Dev 1993, 42(1-2):49-58.
• [18]Cragg GM, Newman DJ: Plants as a source of anti-cancer agents. J Ethnopharmacol 2005, 100(1):72-79.
• [19]Babu TD, Kuttan G, Padikkala J: Cytotoxic and anti-tumor properties of certain types of Umbelliferae with special reference to Centella asiatica (L.) Urban. J Ethnopharmacol 1995, 48(1):53-57.
• [20]Huang YH, Zhang SH, Zhen RX: Asiaticoside inducing apoptosis of tumor cells and enhancing anti-tumor activity of vincristine. Display Settings 2004, 23(12):1599-1604.
• [21]Park YK, Kim Y, Park E, Kim JS, Kang M-H: Estimated flavonoids intake in Korean adults using semiquantitative food frecuency questionnaire. The Korean Journal of Nutrition 2002, 35(10):1081-1088.
• [22]Konoshima T, Takasaki M, Tokuda H, Masuda K, Masuda Y, Arai Y, Shiojima K, Ageta H: Anti-tumor-promoting activities of triterpenoids from ferns. I. Biol Pharm Bull 1996, 19(7):962-965.
• [23]Cha HJ, Bae SK, Lee HY, Lee OH, Sato H, Seiki M: Anti-invasive activity of ursolic acid correlates with the reduced expression of matrix metalloproteinase-9 (MMP-9) in HT1080 human fibrosarcoma cell. Cancer Res 1996, 56(10):2281-2284.
• [24]Jew SS, Yoo CH, Lim DY, Kim H, Mook-Jung I, Jung MW: Structure activity relationship study of Asiatic acid derivatives against beta amyloid (A beta)-induced neurotoxity. Bioorg Med Chem Lett 2000, 10(2):119-121.
• [25]Lee MK, Kim SR, Sung SH, Lim D, Kim H, Choi H: Asiatic acid derivatives protect cultured cortical neurons from glutamate-induced excitotoxicity. Res Commun Mol Pathol Pharmacol 2000, 108(1-2):701-708.
• [26]Babykutty S, Padikkala J, Sathiadevan PP, Vijayakurup V, Azis TKA, Srinivas P, Gopala S: Apoptosis induction of Centella asiatica on human breast cancer cells. Afr J Tradit Complement Altern Med 2009, 6(1):9.
• [27]Cai Y, Lua Q, Sun M, Corke H: Antioxidant activity and phenolic compounds of 112 traditional Chinese medicinal plants associated with anticancer. Life Sci 2000, 74(17):2157-2184.
• [28]Park YK, Park E, Kim J-S, Kang M-H: Daily grape juice consumption reduces oxidative DNA damage and plasma free radical levels in healthy Koreans. Mutation Research/Fundamental and Molelcular Mechanism in Mutagenesis 2003, 529(1-2):77-86.
• [29]Geran R: Protocols for screening chemical agents and natural products against animal tumors and other biological systems. Cancer Chemother Rep 1972, 3:51-61.
• [30]Magnet KJ, Orr MS, Cleveland JL, Rodriguez-Galindo C, Yang H, Yang C, Di Y-M, Jain PT, Gewirtz DA: Suppression of c-myc expression and c-Myc function in response to sustained DNA damage in MCF-7 breast tumor cells. Biochem Pharmacol 2001, 62(5):593-602.
• [31]Fornari FA Jr, Jarvis WD, Grant S, Orr MS, Randolph JK, White F, Mumaw VR, Lovings ET, Freeman RH, Gewirtz DA: Induction of differentiation and growth arrest associated with nascent (nonoligosomal) DNA fragmentation and reduced c-myc expression in MCF-7 human breast tumor cells after continuous exposure to a sublethal concentration of doxorubicin. Cell Growth Differ 1994, 5(7):723.
• [32]Landay M, Oster SK, Khosravi F, Grove LE, Yin X, Sedivy J: Promotion of growth and apoptosis in c-myc nullizygous fibroblasts by other members of the myc oncoprotein family. Cell Death Differ 2000, 7(8):697-705.
• [33]Li D, Tonya M, Shinji S: AFP-L3: a new generation of tumor marker for hepatocellular carcinoma. Clinica Chemica Acta 2001, 313(1-2):15-19.
• [34]Kalra N, Kumar V: C-fos is a mediator of the c-myc-induced appoptitic signaling in serum-deprived hepatoma cells via the p38 mitogen-activated protein kinase pathway. J Biol Chem 2004, 279(24):25313-25319.
• [35]Thompson EB: The many roles of c-Myc in apoptosis. Annu Rev Physiol 1998, 60(1):575-600.
• [36]Dong JM, Naito Y, Tsuruo T: C-myc play a role in cellular susceptibility to death receptor-mediated and chemotherapy-induced apoptosis in human monocytic leukemia U937 cells. Oncogene 1997, 15(6):639-647.
• [37]Tiberio L, Maier JAM, Schiaffonati L: Down-modulation of c-myc expression by phorbol ester protects CEM T leukaemia cells from starvation-induced apotosis: role of ornithine decarboxlase and polyamines. Cell Death Differ 2001, 8(10):967-976.
• [38]Lackinger D, Kaina B: Primary mouse fibroblasts deficient for c-fos, p53 or for both proteins are hypersensitive to UV light and alkylating agent-induced chromosomal breakage and apoptosis. Mutat Res 2000, 457(1-2):113-123.
• [39]Hu L, Hatano M, Ruther U, Tokuhisa T: Overexpression of c-Fos induces apoptosis of CD43+ pro-B cells. J Immunol 1996, 157(9):3804-3811.
• [40]Inada K, Okada S, Phuchareon J, Hatano J, Sugimoto T, Moriya H, Tokuhisa T: C-Fos induces apoptosis in germinal center B cells. J Immunol 1998, 161(8):3853-3861.
• [41]Kaina B, Haas S, Kappes H: A general role for c-fos in cellular protection against DNA-damaging carcinogens and cytostatic drugs. Cancer Res 1997, 57(13):2721-2731.
• [42]Robson S, Ward L, Brown H, Turner H, Hunter E, Pelengaris S, Khan M: Deciphering c-MYC-regulated genes in two distinct tissues. BMC Genomics 2011, 12(1):476. BioMed Central Full Text