CYTOTOXICITY, ANTIPROLIFERATIVE ASSAYS, AND EXPRESION OF P53 AND BCl2 OF ETHANOLIC FRACTION FROM TEA (Camellia sinensis (L.) O.K.) LEAVES INFUSE TO HeLa CELLS

https://doi.org/10.22146/tradmedj.8017

Laela Hayu Nurani(1*)

(1) Faculty of Pharmacy, University of Ahmad Dahlan
(*) Corresponding Author

Abstract


Tea (Camellia sinensis  (L.) O.K.)  is one of medical plant traditionally used by society as anticancer. The aim of this research is to evaluate the citotoxic and antiproliferative effect of ethanolic fraction of tea (Camellia sinensis  (L.) O.K.) and its effect on P53 and bcl-2. Tea were collected and extracted by infundation and fractioned with ethanol as solvent. Citotoxicity test was done by incubating HeLa cell at a density of 2.104 with treatment using extract in concentration of 250; 125 ; 62,5; 31,25; 15,63; and 7,81 µg/mL during 24 hours.  Antiproliferative test were done by calculating doubling time which was equaling a life cell on treatment sample concentration 31,25 µg/mL; 15,63 µg/mL; 7,81 µg/mL; and 3,91 µg/mL with cell control at 24, 48 and 72 hours.  Imunohistochemistry assay was used in LC50  with p53 and bcl-2 antibody. P53 and bcl-2 expression is compared to control. The result indicated that ethanol fraction of tea (Camellia sinensis  (L.) O.K.) leaf infuse has citotoxicity effect on HeLa cell with LC50 of 24,45 µg/mL. The result of  doubling time test indicated that doubling time value for a solvent  control is 74,11 hours and a cell control is 78,22 hours. While, with treatment 31,25 µg/mL; 15,63 µg/mL; 7,81 µg/mL; and 3,91 µg/mL resulted on negative slope then did not produce doubling time. The extract can induce p53 expression and inhibit bcl-2  expression compared to control.


Keywords


Camellia sinensis, HeLa, p53, bcl-2



References

Akroum, S., Haddi, M.L., Lalaoui, 2009, Fungal tannase degrading condensed tannins of Camellia sinensis and measure of the enzyme activity on quebracho, J. of Scientific Research 4(4):237-241

Akroum, S., Satta, D., Lalaoui, K., 2009, Antimicrobial, Antioxidant, Cytotoxic Activities and Phytochemical Screening of Some Algerian Plants, Eur. J. of Scientific Research, 31(2):289-295

Antonsson, B., and Martinou, J.C., 2000, The Bcl-2 Protein Family, Experimental Cell Research, 256, 50-57

Azis, 2009, Gynecological cancer in Indonesia, J Gynecol Oncol , 20(1):8-10

Canavan TP, Doshi NR. 2000, Cervical cancer. Am Fam Physician, 61:1369-76

Chen, L., Zhang, H.Y., 2007, Cancer preventive mechanisms of teh green tea polyphenol-epigallocatechin-3-gallate. Molecules, 3:12 (5) :946-57

Field, S.J., F.Y., Kuo, F., Zubiaga, A.M., Kaelin, Jr, W.G., Livingston, D.M., Orkins, S.H., Greenberg, M.E., 1996, E2F-1, Function in Mice to Promote Apoptosis and Supress Proliferation, Cell, 85: 549-561

Hemann, M.T., Lowe S.W., 2006, The p53-Bcl-2 connection, Cell Death and Differentiation 13, 1256-1259

Hongda Z., 1994, The anticancer effect and anti-DNA topoisomerase II effect of extracts of camellia ptilophylla chang and camellia sinesis, Medicine Chinese Journal of Cancer Research, 6(3):184-190

Jerry, D., J., Minter, M.L., Becker, K.A., Blackburn, A.C., 2002, Hormonal control of p53 and chemoprevention, Breast Cancer Res, 4: 91-94

Li W.G., Li, Q.H., Tan, Z., 2005, Epigallocatechin gallate induces telomere fragmentation in HeLa and 293 but not in MRC-5 cells, Life Sc., 76(15):1735-1746

Nishimura, A., Nakahara, T., Ueno, T., Sasaki, K., Yoshida, S., Kyo, S., Howley, P.M., and Sakai, H., 2006, Requirement of E7 oncoprotein for viability of HeLa cells, , 8: 984-993

Nurrochmad, A., 2001, Sintesis Kurkumin, Bisdemetoksi Kurkumin, Bisdemetoksi- dehidroksikurkumin dan Pentagamavunon- O serta Uji Ketoksikannya terhadap Sel Meiloma dan Sel Mononuklear Normal secara In Vitro, Tesis, Program Pasca Sarjana Universitas Gadjah Mada, Yogyakarta.2004, Flavonoid biosyntehsis in teh tea plant Camellia sinensis: properties of enzymes of teh prominent epicatechin and catechin pathways, Arch Biochem Biophys, 431(1):22-30

Ravindranath, M.H, Saravanan, T.S., Monteclaro1, C.C., Presser N., Xing Ye, X., Selvan, S.R., and Brosman, S, 2006, Epicatechins Purified from Green Tea (Camellia sinensis), Differentially Suppress Growth of Gender-Dependent Human Cancer Cell Lines, Advance Access Publication, 3(2)237-47

Sigma, 1999, Biochemicals and Reagents for Life Science Research, Sigma Aldrich CO, Singapore, 1873

Teissier, S., Pang, C.L., and Thierry, F., 2010, The E2F5 repressor is an activator of E6/E7 transcription and of the S-phase entry in HPV18-associated cells, Oncogene, 29(36):5061-70

Yao, L., Caffin, N., Darcy, B., Jiang, Y., Shi, J., Singanusong, R., Liu, X., Datta, N., Kakuda, Y., Xua, Y., 2005, Seasonal Variations of Phenolic Compounds in Australia-Grown Tea (Camellia sinensis), J. Agric. Food Chem., 53 (16), pp 6477–6483



DOI: https://doi.org/10.22146/tradmedj.8017

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