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. 2012 Aug 22:12:134.
doi: 10.1186/1472-6882-12-134.

In vitro evaluation of Pandanus amaryllifolius ethanol extract for induction of cell death on non-hormone dependent human breast adenocarcinoma MDA-MB-231 cell via apoptosis

Hueh Zan Chong  1 , Swee Keong Yeap, Asmah Rahmat, Abdah Md Akim, Noorjahan Banu Alitheen, Fauziah Othman, Cheng Lian Gwendoline-Ee
Affiliations

In vitro evaluation of Pandanus amaryllifolius ethanol extract for induction of cell death on non-hormone dependent human breast adenocarcinoma MDA-MB-231 cell via apoptosis

Hueh Zan Chong et al. BMC Complement Altern Med. .

Abstract

Background: Our previous study had shown that P. amaryllifolius was able to selectively inhibit cell proliferation of hormone independent breast cancer cell line MDA-MB-231. To understand the mode of killing and mechanism of action for P. amaryllifolius, the ethanol extract was evaluated for their alteration of cell cycle progression, PS externalization, DNA fragmentation and expression of anti/pro-apoptotic related protein.

Results: Cell cycle progression analysis, Annexin V and Tunel assays suggested that IC50 of P. amaryllifolius ethanol extract induced G0/G1 cell cycle arrest, PS externalization and DNA fragmentation. On the other hand, ELISA for cytochrome c, caspase-3/7, 8 and 9 indicated that apoptosis was contributed by mitochondrial cytochrome c release via induction of caspase 3/7, 9, and p53 was associated with the suppression of XIAP in P. amaryllifolius treated MDA-MB-231 cells.

Conclusion: Our findings suggest that P. amaryllifolius ethanol extract induced apoptosis on hormone independent breast cancer cell line MDA-MB-231.

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Figures

Figure 1
Figure 1
Effects of P. amaryllifolius extract on MDA-MB-231 cell DNA fragmentation. ( a) P. amaryllifolius altered cell cycle distributions (%) but not sub-G0/G1 population in treated breast cancer MDA-MB-231 cells as compared to controls after 24 hr of treatment in cell cycle PI study. ( b),( c) P. amaryllifolius induced subG0/G1 population of MDA-MB-231 cell after 48 and 72 hr treatment exposure detected by cell cycle PI staining assay. ( d) P. amaryllifolius increased cell population of early and secondary apoptosis after 48 hr of treatment in AnnexinV/PI study. ( e), ( f) P. amaryllifolius induced DNA fragmentation of MDA-MB-231 cell after 48 and 72 hr treatment exposure detected by TUNEL assay. Values are presented as means (n = 3) ± S.E. *signified (p < 0.05).
Figure 2
Figure 2
Pandanus amaryllifolius extract induced translocation of cytochrome c into the cytosol, activation of initiator caspase 8, 9 and effector caspase 3/7 (fold change) in treated MDA-MB-231 cells as compared to control cells. Values are presented as means (n = 3) ± S.E. *signified (p < 0.05).
Figure 3
Figure 3
P. amaryllifolius extract induced the population of MDA-MB-231 cell that expressed tumour suppressor p53 protein while downregulated the population that expressed apoptosis inhibitor protein XIAP as compared to control cells. Values are presented as means (n = 3) ± S.E. *signified (p < 0.05).
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