Apoptotic Activity of Aporphine from Stephania venosa on Human Ovarian Cancer CellsK. Montririttigri, P. Moongkarndi,* S. Joongsomboonkusol, B. Chitkul and K. Pattanapanyasat
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Stephania venosa is a plant rich of alkaloids of family Menispermaceae. In Thailand, the study on this plant revealed a wild variety of phytochemical constituents. Those including flavonoids, alkaloids, terpenoids, sulfides, and polyphenolics. In this study, we analyzed the anticancer activity according to the antiproliferation and apoptotic activity of the pure constituent isolated from S. venosa tuber on human ovarian cancer cells (SKOV3). During phytochemical processes, we concomitantly tested the activity of the isolates and selected the effective fractions for further purification. Crude ethanolic extract and pure constituent were obtained from the tuber of S. venosa and evaluated the cytotoxic activity against SKOV3 by MTT assay. Crude ethanolic extract was performed various fractionations and further isolated to obtain pure constituent. Purified constituent and crude extract were demonstrated the significant effect on antiproliferation in a dose dependent manner. Apoptotic effect was confirmed by morphological changes, DNA fragmentation and caspase activation assay. The cytotoxic activity of crude ethanolic extract showed a potent inhibition with an ED50 value at 31 µg/ml. Structure of the pure constituent was determined by NMR technique and identified as aporphine. Aporphine from S. venosa showed the antiproliferation at ED50 value at 6 µg/ml against SKOV3. Results from the morphological changes, DNA fragmentation and caspase activation assay, in this study demonstrated that aporphine could significantly inhibit the treated tumor cell proliferation and cause cells death via apoptosis, whereas those were not found in the untreated cells. We concluded that the pure constituent from S. venosa could lead to the further study on the detail mechanisms of action of this active substance for future application as new drug for ovarian cancer.
Keyword:
alkaloids, apoptosis, aporphine, cancer, caspase, DNA fragmentation, MTT assay, NMR, Stephania venosa, Ovarian Cancer
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