Morphological studies of apoptotic HeLa cells death induced by eurycomanone

Nurkhasanah ., Azimahtol Hawariah Lope Pihie, Jalifah Latip


Eurycomanone is a cytotoxic ingredient found in Eurycoma longifolia Jack. Previous studies have noted its activity against many epithelial cell lines. In this study, eurycomanone had obvious cytotoxic effect on HeLa cells by methylene blue staining assays. After HeLa cells were treated with eurycomanone, typical morphological changes, including cytoplasm shrinkage and decrease of cell volume were observed by light microscope. Chromatin condensation and nuclear fragmentation could be observed by fluorescence microscope after staining with Hoechst 33258 nuclear staining. DNA fragmentation and apoptotic body formation that is characteristic of apoptosis could be determined after treated cells were assayed with TUNEL (terminal deoxynucleotidyl transferase mediated d-UTP nick end labeling). The externalization of phosphatidyl serine (PS) could be determined by flow cytometry analysis using Annexin-V/PI double staining. The result suggested that eurycomanone exerted antiproliferative activity on HeLa cells by inducing apoptosis.

Key words : eurycomanone, cytotoxic, apoptosis, HeLa


Allen, R. T., Hunter III, W. J., and Agrawal, D. K., 1997, Morphological and biochemical characterization and analysis of apoptosis. J. of Pharmacological and Toxicological Methods 37, 215-228.

Brown, L.R. 2003. Laboratory exercise: A laboratory exercise in comparative DNA analysis. Biochem. Mol. Biol. Edu. 31, 177-179.

Chan, K. L., O’Neill, M. J., Phollopson, J. D., and Warhurst, D. C. 1986. Plant as source of antimalarial drugs. Planta Medica 52: 105-107.

Cheah, S. C., and Azimahtol, H. L. P. 2004. Eurycomanone exert antiproliferative activity via apoptosis upon MCF-7 cells. Prosiding symposium Biologi Kebangsaan Malaysia ke-7. 73-77.

Darise, M., Kohda, H., Mizutani, K., and Tanaka, O. 1982. Eurycomanone and eurycomanol, quassinoids from the roots of Eurycoma longifolia. Phytochemistry, 21, 2091-2093.

Dibartolomeis, S. M., and Monė, J. P. 2003. Apoptosis: a four week laboratory investigation for advanced molecular and cellular biology students. Cell Bio.l Edu. 7242F, 275-295.

Fadok, V. A., and Henson, P. M. 2003. Apoptosis: Giving phosphatidylserine recognition an assist-with a twist. Current Biology 13, R655-R657.

Franco, E. L., Schlecht, N. F., and Saslow, D. 2003. The epidemiology of cervical cancer. Cancer Journal 9, 348-359.

Frankfurt, O. S., and Krishan, A. 2003. Apoptosis-based drug screening and detection of selective toxicity to cancer cells. Anticancer Drugs 14, 555-561.

Gewies, A. 2003. Introduction to Apoptosis. Apo.Review. 1-26.

Hishikawa, K., Oemar, B. S., Tanner, F. C., Nakaki, T., Luscher, T. F., and Fujii, T. 1999. Connective tissue growth factor induces apoptosis in human breast cancer cell line MCF-7. J. Biol. Chem. 274, 37461-37466.

Kardono, L. B. S., Angerhofer, C. K., Tsauri, S., Padmawinata, K., Pezzuto, J. M., and Kinghorn, A.D. 1991. Cytotoxic and antimalarial constituents of the roots of Eurycoma longifolia, J. Nat. Prod. 54, 1360-1367.

Kerr, J. F. R., Winterford, C. M., and Harmon, B. V. 1994. Apoptosis its significance in cancer and cancer therapy. Cancer 73, 2013-2026.

Kuo, P. C., Damu, A. G., Lee, K. H., and Wu, T. S. 2004. Cytotoxic and antimalarial constituents from the roots of Eurycoma longifolia. Bioorg. Med. Chem. 12, 537-544.

Lawen, A. 2003. Apoptosis-an introduction. BioEssays 25, 888-896.

Lee, A. T. C., and Azimahtol, H. L. P. 2003. Stryrylpyrone derivative induces apoptosis through the up- regulation of Bax in the human breast cancer cell lines. J. Biochem. Mol. Biol. 36, 269-274.

Lin, L., and Hwang, P. L. 1991. Antiproliferative effect of oxygenated sterols: positife correlation with binding affinities for the antiestrogen binding sites. Biochim. Biophys. Act. 1082, 177-184.

Oka, Y., Naomoto, Y., Yasuoka, Y., Hatano, H., Haisa, M., Tanaka, N., and Orita, K. 1997. Apoptosis in cultured human colon cancer cells induced by combined treatments with 5-fluorouracil, tumor necrosis factor-α and interferon-α. Jpn. J. Clin. Oncol. 27, 231-235.

Reed, J. C. 2001. Apoptosis-regulating proteins as targets for drug discovery. Trends in Molecular Medicine 7, 314-319.

Savill, J., and Fadok, V. 2000. Corpse clearance defines the meaning of cell death. Nature 407, 784-788.

Song, X. K., and Zhang, W. 2006. Growth inhibition effect of parthenolide on human hepatocellular carcinoma cell line BEL-7402. Chem. Res. Chinese U. 22, 173-176.

Tee, T. T., and Azimahtol, H.L.P. 2005. Induction of apoptosis by Eurycoma longifolia Jack extracts. Anticancer Res. 25, 2205-2214.

Wall, M. E., Taylor, H., and Wani, M. C. R. 1987. Plants antitumor agents 24 rapids 9-KB assay. J. Nat. Prod. 50, 764-766.



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