Main Article Content
Epipremnum pinnatum (L.) Engl. Chloroform extract produced significant growth inhibition against T-47D breast carcinoma cells and analysis of cell death mechanism indicated that the extract elicited both apoptotic and non-apoptotic programmed cell deaths. T-47D cells exposed to the extract produced a significant up-regulation of c-myc and caspase-3 mRNA expression levels as compared to untreated cells. The up-regulation of caspase-3 mRNA expression appeared to be mediated mainly via both protein kinase C and tyrosine kinases pathways. T-47D cells exposed to the extract at EC50 concentration (72 h) for 24 h demonstrated typical DNA fragmentation associated with apoptosis, as carried out using a DNA fragmentation detection assay. However, ultrastructural analysis using transmission electron microscope demonstrated distinct vacuolated cells, which indicated a Type II non-apoptotic cell death although the presence of cell and nuclear blebbing, apoptotic bodies and chromatin changes associated with apoptosis were also detected. The presence of non-apoptotic programmed cell death was also detected with annexin-V and propidium iodide staining. These findings suggested that up-regulation of caspase-3 and c-myc mRNA expression may have contributed to both apoptotic and non-apoptotic programmed cell death, respectively in the Epipremnum pinnatum (L.) Engl. Chloroform extract-treated T-47D cells.
Keywords: Medicinal Plant, Epipremnum pinnatum (L.) Engl., apoptotic, non-apoptotic
Corresponding author: E-mail: firstname.lastname@example.org
Copyright Transfer Statement
The copyright to this article is transferred to Current Applied Science and Technology journal (CAST) with effect if and when the article is accepted for publication. The copyright transfer covers the exclusive right to reproduce and distribute the article, including reprints, translations, photographic reproductions, microform, electronic form (offline, online) or any other reproductions of similar nature.
The author warrants that this contribution is original and that he/she has full power to make this grant. The author signs for and accepts responsibility for releasing this material on behalf of any and all co-authors.
 Wong, K.T., Tan, B.K.H., Sim, K.Y., Goh, S.H. 1996. A cytotoxic melanin precursor, 5,6-Dihydroxyindole, from the folkloric anti-cancer plant; Rhaphidophora korthalsii, Natural Product Letters, 9(2), 137-140.
 Chan, M.J., Turner, I.M. 1998. The use of Epipremnum pinnatum (Araceae) in Singapore in the treatmeant of cancer: an unreported application of a herbal medicine, Economic Botany, 52(1), 108.
 Goh, S.H. 1999. Bioactive principles from folkloric anti-neoplastic plants and from bio-prospecting the Malaysian Forest. In: The International Conference on Biodiversity and Bioresources: Conservation and Utilization, Phuket, Thailand, November, 23-27 (http://www.iupac.org/symposia/proceedings/phuket/goh.html)
 Tan, M.L.,Najimudin, N., Sulaiman, S.F., Tengku Muhammad, T.S. 2005. Growth arrest and non-apoptotic programmed cell death associated with the up-regulation of c-myc mRNA expression in T-47D breast tumor cells following exposure to Epipremnum pinnatum (L.) Engl. Hexane extract, Journal of Ethnopharmacology., 96, 375-383.
 Raskin, I., Ribnicky, D.M., Komarnytsky, S., Ilic, N., Poulev, A., Borisjuk, N., Brinker, A., Moreno, D.A., Ripoll, C., Yakoby, N., O’ Neal, J.M., Cornwell, T., Pastor, I., Fridlender, B. 2002. Plants and human health in the twenty first century, Trends in Biotechnology, 20(12), 522-531.
 Kibertis, P., Roberts, L. 2002. It’s not just the genes. Science, 296, 685.
 Gryfe, R., Swallow, C., Bapat, B., Redston, M., Gallinger, S., Couture, J. 1997. Molecular biology of colorectal cancer, Current Problems in Cancer, 21, 233-300.
 Potten, C.S. 1997. Epithelial cell growth and differentiation. American Journal of Physiology, 36, G253-G257.
 Kamesaki, H. 1998. Mechanisms involved in chemotherapy-induced apoptosis and their implications in cancer chemotherapy, International Journal of Hematology, 68, 29-43.
 Arends, M.J., Wyllie, A.H. 1991. Apoptosis mechanisms and roles in pathology, International Review of Experimental Pathology, 32, 223-54.
 Kerr, J.F.R.,Winterford, C.M., Harmon, B.V. 1994. Apoptosis; its significance in cancer and cancer therapy, Cancer, 73(8), 2013-26.
 Gavrieli, Y., Sherman, Y., Ben-Sasson, S.A. 1992. Identification of programmed cell death in-situ via specific labeling of nuclear DNA fragmentation, Journal of Cell Biology, 119(3), 493-501.
 Fadok, V.A., Savill, J.S., Haslett, C., Bratton, D.L., Doherty, D.E., Campbell, P.A., Henson, P.M. 1992(a). Different populations of macrophages use either the vitronectin receptor or the phosphatidylserine receptor to recognize and remove apoptosis, Journal of Immunology, 149(12), 4029-35.
 Fadok, V.A., Voelker, D.R., Campbell, P.A., Cohen, J.J., Bratton, D.L., Henson, P.M. 1992b. Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages, Journal of Immunology, 148, 2207-16.
 Verhoven, B., Schlegel, A.R., Williamson, P. 1995. Mechanism of phosphatidylserine exposure, a phagocyte recognition signal, on apoptotic T-lymphocytes, Journal of Experimental Medicine, 182, 1597-1601.
 Vermes, I., Haanem, C., Steffens-Nakken, H., Reutelingsperger, C. 1995. A novel assay for apoptotic cells using fluorescein labelled annexin V, Journal of Immunological Methods, 184, 39-51.
 Adams, J.M., Cory, S 2002. Apoptosomes: engines for caspase activation, Current Opinion in Cell Biological, 14, 715-720.
 Beauparlant, P., Shore, G.C. 2003. Therapeutic activation of caspases in cancer: a question of selectivity, Current Opinion in Drug Discovery and Development, 6, 179-187.
 Kastan, M.B., Onyekwere, O., Sidransky, D., Vogelstein, B., Craig, R.W. 1991. Participation of p53 protein in the cellular response to DNA damage, Cancer Research, 51(23), 6304-11.
 Nelson, W.G., Kastan, M.B. 1994. DNA strand breaks: the DNA template alterations that trigger p53-dependent DNA damage response pathways, Molecular Cell Biology, 14(3), 1815-23.
 Kim, R., Ohi, Y., Inoue, H., Toge, T. 1999. Taxotere activates transcription factor AP-1 in association with apoptotic cell death in gastric cancer cell lines, Anticancer Research, 19(16B), 5399-405.
 Janus, F., Albrechtsen, N., Dornreiter, I., Wiesmuller, L., Grosse, F., Deppert, W. 1999. The dual role model for p53 in maintaining genomic intergrity, Cellular and Molecular Life Sciences, 55, 12-27.
 Wurm, F.M., Gwinn, K.A., Kingston, R.E. 1986. Inducible overproduction of mouse c-myc protein in mammalian cells, Proceedings of the National Academy of Sciences USA, 83, 5414-8.
 Wyllie, A.H., Rose, K.A., Morris, R.G., Steel, C.M., Foster, E., Spandidos, D.A. 1987. Rodent fibroblast tumours expressing human myc and ras genes; growth, metastasis and endogenous oncogene expression, British Journal of Cancer, 56, 251-9.
 Evan, G.I., Wyllie, A.H., Gilbert, C.S., Littlewood, T.D., Land, H., Brooks, M., Waters, C.M., Penn, L.Z., Hancock, D.C. 1992. Induction of apoptosis in fibroblasts by c-myc protein, Cell, 69, 119-28.
 Askew, O.S., Ashmun, R.A., Simmons, B.C., Cleveland, J.L. 1991. Constitutive c-myc expression in an IL-3-dependent myeloid cell line suppresses cell cycle arrest and accelerates apoptosis, Oncogene, 6, 1915-22.
 Kuida, K., Zheng, T.S., Na, S.Q., Kuan, C.Y., Yang, D., Karasuyama, H., Rakic, P., Flavell, R.A. 1996. Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice, Nature, 384, 368-72.
 Schlegel, J., Peter, I., Orrenius, S., Miller, D.K., Thornberry, N.A., Yamin, T.T., Nicholson, D.W. 1996. CPP-32/apopain is the key interleukin-1β-converting enzyme-like protease involved in Fas-mediated apoptosis, Journal Biological Chemistry, 271(4), 1841-4.
 Janicke, R.U., Sprengart, M.L., Wati, M.R., Porter, A.G. 1998. Caspase-3 is required for DNA fragmentation and morphological changes associated with apoptosis, Journal of Biological Chemistry, 273(16), 9357-60.
 Kousteni, S., Tura-Kockar, F., Ramji, D.P. 1999. Sequence and expression analysis of a novel Xenpus laevis cDNA that encodes a protein similar to bacterial and chloroplast ribosomal protein L24, Gene, 235, 13-18.
 Tengku Muhammad, T.S., Hughes, T.R., Ranki, H., Cryer, A., Ramji, D.P. 2000. Differential regulation of macrophage CCAAT-Enhancer binding protein isoforms by lipopolysaccharide and cytokines, Cytokine, 12, 1430-1436.
 Ahn, C.H., Kong, J.Y., Choi, W.C., Hwang, M.S. 1996. Selective inhibition of the effects of phorbol ester on doxorubicin resistance and p-glycoprotein by the protein kinase C inhibitor 1-(5-isoquinolinesulfonyl)-2-methylpiperazine (H7) in multidrug-resistant MCF-7/Dox human breast carcinoma cells, Biochemical Pharmacology, 52, 393-9.
 Lin, C.H., Kuan, I.H., Wang, C.H., Lee, H.M., Lee, W.S., Sheu, J.R., Hsiao, G., Wu, C.H., Kuo, H.P. 2002. Lipoteichoic acid-induced cyclooxygenase-2 expression requires activations of p44/42 and p38 mitogen-activated protein kinase signal pathways, European Journal of Pharmacology, 450, 1-9.
 Fujihara, M., Muroi, M., Muroi, Y., Ito, N., Suzuki, T. 1993. Mechanism of lipopolysaccharide-triggered junB activation in a mouse macrophage-like cell line (J774), Journal of Biological Chemistry, 268(20), 14898-905.
 Eason, S., Martin, W. 1995. Involvement of tyrosine kinase and protein kinase C in the induction of nitric oxide synthase by lipopolysaccharide and interferon-gamma in J774 macrophages, Archives International Pharmacodynamic Therapy, 330(2), 225-40.
 Glass, D.B. 1983. Differential responses of cyclic GMP-dependent and cyclic AMP-dependent protein kinases to synthetic peptide inhibitors, Biochemical Journal, 213, 159-64.
 Tengku-Muhammad, T.S., Hughes, T.R., Cryer, A., Ramji, D.P. 1999. Involvement of both the tyrosine kinase and the phosphatidylinositol-3’ kinase signal transduction pathways in the regulation of lipoprotein lipase expression in J774.2 macrophages by cytokines and lipopolysaccharide, Cytokine, 11(7), 463-8.
 Geran, R.I., Greenberg, N.H., Macdonald, M.M., Schumacher, A.M., Abbott, B.J. 1972. Protocols for screening chemical agents and natural products against animal tumours and other biological systems, Cancer Chemotherapy Reports, 3, 59-61.
 Hoffman, B., Liebermann, D.A. 1998. The proto-oncogene c-myc and apoptosis, Oncogene, 17, 3351-7.
 Leng, Y., Gu, Z.P., Cao, L. 2000. Apoptosis induced by droloxifene and c-myc, bax and bcl-2 mRNA expression in cultured luteal cells of rats, European Journal of Pharmacology, 409(2), 123-31.
 Thornberry, N.A., Lazebnik, Y. 1998. Caspases: enemies within, Science, 281, 1312-1316.
 Budihardjo, I., Oliver, H., Lutter, M., Luo, X., Wang, X. 1999. Biochemical pathways of caspase activation during apoptosis, Annual Review of Cell and Development Biology, 15, 260-290.
 Wolf, B.B., Green, D.R. 1999. Suicidal tendencies: apoptotic cell death by caspase family proteinases, Journal Biological Chemistry, 274, 20049-20052.
 Earnshaw, W.C., Martins, L.M., Kaufmann, S.H. 1999. Mammalian caspases: Structure, activation, substates and functions during apoptosis, Annual Review of Biochemistry, 68, 383-424.
 Blatt, N.B., Glick, G.D. 2001. Signaling pathways and effector mechanisms pre-programmed cell death, Bioorganic and Medicinal Chemistry, 9, 1371-84.
 Chang, Q., Tepperman, B.L. 2001. The role of protein kinase C isozymes in TNF-alpha-induced cytotoxicity to a rat intestinal cell line, American Journal of Physiology – Gastrointestinal and Liver Physiology, 280(4), G572-83.
 Basu, A., Akkaraju, G.R. 1999. Regulation of caspase activation and cis-diammine-dichloroplatinum(II)-induced cell death by protein kinase C, Biochemical Journal, 38(14), 4245-51.
 Park, I.C., Park, M.J., Rhee, C.H., Lee, J.I., Choe, T.B., Jang, J.J., Lee, S.H., Hong, S.I. 2001. Protein kinase C activation by PMA rapidly induces apoptosis through caspase-3/CPP32 and serine protease(s) in a gastric cancer cell line, International Journal of Oncology, 18(5), 1077-83.
 Cataldi, A., Miscia, S., Centurione, L., Rapino, M., Bosco, D., Grifone, G., Valerio, V.D., Garaci, F., Rana, R. 2002. Role of nuclear PKC delta inmediating caspase-3 up-regulation in Jurkat T leukemic cells exposed to ionizing radiation, Journal of Cellular Biochemistry, 86(3), 553-60.
 Koriyama, H., Kouchi, Z., Umeda, T., Saido, T.C., Momoi, T., Ishiura, S., Suzuki, K. 1999. Proteolytic activation of protein kinase C δ and ε by caspase-3 in U937 cells during chemotherapeutic agent-induced apoptosis, Cell Signaling, 11(11), 831-38.
 Sato, N., Sakamaki, K., Terada, N., Arai, K., Miyajima, A. 1993. Signal transduction by the high-affinity GM-CSF receptor: two distinct cytoplasmic regions of the common beta subunit responsible for different signaling, EMBO Journal, 12(11), 4181-9.
 Bowen, I.D., Bowen, S.M. 1990. Programmed cell death in tumours and tissues, 1st edition, Chapman and Hall, London.
 Bowen, I.D., Mullarkey, K., Morgan, S.M. 1996. Programmed cell death in the salivary glands of blow fly Calliphora vomitoria, Microscopy Research and Techniques, 34, 202-7.
 Bowen, I.D., Bowen, S.M., Jones, A.H. 1998. Mitosis and Apoptosis: Matters of life and death, Chapman and Hall, London.
 Bowen, I.D., Amin, F. 2000. Tumor cell death. In: Programmed cell death in animals and plants (eds Bryant et al), BIOS Scientific Publishers, United Kingdom.
 Amin, F., Bowen, I.D., Szegedi, Z., Mihalik, R., Szende, B. 2000. Apoptotic and non-apoptotic modes of programmed cell death in MCF-7 human breast carcinoma cells, Cell Biology International, 24(4), 253-60.
 Szende, B., Keri, G.Y., Szegedi, Z.S., Benedeczky, I., Csikos, A., Orfi, L., Gazit, A. 1995. Tryphostin induces non-apoptotic programmed cell death in colon tumor cells, Cell Biology International, 19, 903-11.
 Ou, D., Bonomi, P., Jao, W., Jadko, S., Harris, J.E., Anderson, K.M. 2001. The mode of cell death in H-358 lung cancer cells cultured with inhibitors of 5-lipooxygenase or the free radical spin trap, NTBN. Cancer Letters, 166(2), 223-31.
 Ellington, A.A., Berhow, M., Singletary, K.W. 2005. Induction of macroautophagy in human colon cancer cells by soybean B-group triterpenoid saponins, Carcinogenesis, 26(1), 159-67.
 Tanida, I., Ueno, T., Kominami, E. 2004. LC3 conjugation system in mammalian autophagy, The International Journal of Biochemistry and Cell Biology, 36, 2503-2518.
 Meijer, A.J., Dubbelhuis, P.F. 2004. Amino acid signaling and the integration of metabolism, Biochemical and Biophysical Research Communications, 313, 397-403.
 Gozuacik, D., Kimchi, A. 2004. Autophagy as a cell death and tumor suppressor mechanism, Oncogene, 23(16), 2891-2906.
 Marino, G., Lopez-Otin, C. 2004. Autophagy: molecular mechanisms, physiological functions and relevance in human pathology, Cellular and Molecular Life Sciences, 61(12), 1439-1454.
 Meijer, A.J., Codogno, P. 2004. Regulation and role of autophagy in mammalian cells, International Journal Biochemistry and Cell Biology, 36(12), 2445-2462.
 Gutierrez, M.G., Munafo, D.B., Beron, W., Colombo, M.I. 2004. Rab 7 is required for the normal progression of the autophagic pathway in mammalian cells, Journal Cell Science, 117(13), 2687-2697.