Alkali hydrolysate of total saponins from Pulsatilla chinensis inhibits hepatic carcinoma cell growth by inducing apoptosis through mitochondrial pathway / 中草药

ABSTRACT

Objective: To investigate the anti-hepatocarcinoma effect and mechanisms of the alkali hydrolysate of total saponins from Pulsatilla chinensis (PAHS). Methods: MTT assay was used to evaluate the effect of PAHS on proliferation of human liver cancer cell line SMMC-7721 in vitro; Cell morphology was observed by Giemsa staining. The effects of PAHS on apoptosis, cell cycle, and mitochondrial membrane potential of SMMC-7721 were detected by Hoechst 33258 staining and flow cytometry assay; Western blotting was employed to detect the protein expression of Cytochrome C, Caspase-3, cleaved Caspase-3, and Bcl-2 in SMMC-7721 cells. An in vivo liver cancer model was established using ICR mice subcutaneously received H22 hepatoma carcinoma cells to detect tumor growth inhibitory rate. Morphological changes of the tumor samples were observed by HE staining and transmission electron microscope. Results: MTT assay showed that PAHS could inhibit proliferation and increase apoptosis of SMMC-7721 cells in dose-and time-dependent manners in vitro, block the cell cycle in S phase, and decrease mitochondria membrane potential; PAHS could significantly increase the expression of Cytochrome C and cleaved Caspase-3 and decrease the expression of Bcl-2 and Caspase-3. PAHS dramatically decreased the weight of tumor tissue in a dose-dependent manner. Histopathological examination showed that large necrosis area and apoptotic cells were found in tumor tissues of mice in PAHS administrated group. Conclusion: PAHS exerts antitumor activity in vitro and in vivo by inducing apoptosis, and its mechanism is related to regulation of the mitochondrial pathway.