Study on Apoptosis Mechanism of Human Colon Cancer Cell Lines HCT-116 Induced by Solasodine

OBJECTIVE To explore the effects of Solasodine on the apoptosis of human colon cancer cell line HCT-116 and the expression of associated genes，and to investigate the mechnism of the inhibition of HCT-116 cells proliferation. METHODS After treatment， the inhibitory effect of HCT-116 cell proliferation was determined by Real time non-label cell analyzer. Flow cytometry was employed to detect cell apoptosis. The qPCR method was used to determine the mRNA levels of Bax and Survivin. Protein levels of Bcl-2， Bax， Bcl-xl， Caspase-3， Caspase-8， Caspase-9， PARP， PI3K and Akt were determined by Western blot . RESULTS Real time non-label cell analysis result showed that Solasodine inhibited the growth of HCT-116 in vitro， and the potency enhanced with the increase of drug dose and treat time. Flow cytometry found that the cell apoptosis increased with the increase of drug concentration. The qPCR showed that the mRNA levels of apoptosis-related genes Bax and Survivin changed. Western blot results revealed that Bax protein levels increased， while Bcl-2 and Bcl-xl protein levels decreased， and the ratio of Bax/Bcl-2 was also increased. It showed that Solasodine also promoted the Caspase-3， Caspase-8， Caspase-9 and PARP activities， and inhibited the activation of PI3K and Akt . CONCLUSION Solasodine can inhibit HCT-116 cells proliferation and induce the cell apoptosis via blocking PI3K/Akt signaling pathway and regulating Caspase family， the Bcl-2 family， and the expression of Survivin and PARP. It can regulate these two apoptotic pathways， mitochondrial pathway and death receptor pathway.