Studies On Antitumor Effects And Mechanisms Of Sasanquasaponin A

Object: Sasanquasaponin A is an anti-tumor active compound which has been extracted and separated from the roots of Camellia oleifera Abel, but its role mechanism of action is unclear. This study was designed to research the anti-liver cancer and the anti-lung cancer mechanisms of sasanquasaponin A in vitro and in vivo. Methods: MTT assay was used to evaluate proliferational inhibition of sasanquasaponin A on human hepatoma cells BEL-7402 and lung cancer A549; At the same time, the morphology of BEL-7402 and A549 tumour cells were also observed by Giemsa staining; ICR mice were used to establish H22 hepatoma carcinoma cell tumor model. The effects of sasanquasaponin A was examined on anti-tumor in vivo; The H22 tumor tissue morphologic were observed by HE staining; The electron microscopy was used to examine tumor structural changes; Hoechst33342 assay kits were used to observe the apoptotic morphology of A549 and BEL-7402 cells; Flow cytometry was used to analyze the apoptosis and the cell cycle of A549, BEL-7402 cells; The electron microscopy was used to examine autophagosome and apoptotic. Western blot method was employed to determine Bcl-2 、 Caspase-3 and autophagy-related protein LC3 etc. expression in BEL-7402 and A549 cells. Results: Sasanquasaponin A could inhibit proliferation of tumor cells by MTT assay in concentration-dependent and time manners; Giemsa staining results showed that sasanquasaponin A induced tumor cells smaller, concentrated nucleus, cell fragmentation; Sasanquasaponin A(1, 2mg/kg) could significantly inhibit the growth of H22 weight(P<0.05, P<0.001), also showed toxicity; Hoechest33342 staining was found with increasing dose, to send a strong blue fluorescence of cells, more and more cells large number of apotosisi; Electron microscopy results there are increasing autophagic cell; Sasanquasaponin A(6.25, 12.50, 18.75μM) could significantly induced apoptosis of A549 and BEL-7402 cells(P<0.01, P<0.001); Sasanquasapon A can block the cell cycle G2 phase in A549 cells and arrest BEL-7402 cells in S phase;Sasanquasaponin A could significantly increased the caspase-3 expression(P<0.01), down-regulated the expression of Bcl-2 in BEL-7402 and A549 cells; Sasanquasaponin A(6.25, 12.50μM) can significantly upregulated autophagy marker proteins LC3-Ⅱ, caspase-3, P53 expression(P<0.05) in BEL-7402 and A549 cells. Conclusion: Sasanquasaponin A shows a favourable anti-tumor effects in vivo, and upregulate LC3-Ⅱ, caspase-3, P53 expression, and its anti-tumor effect and p-53-m TOR mediated autophagy pathway and caspase-3, bcl-2 apoptosis pathway.