积雪草酸联合奥沙利铂对结肠癌HCT116细胞凋亡、自噬和焦亡的调控作用研究

目的:探讨积雪草酸(asiatic acid,AA)联合奥沙利铂(oxaliplatin,OXP)对结肠癌HCT116细胞增殖、凋亡、自噬和焦亡的调控作用.方法:采用CCK8法检测AA、OXP、AA与OXP联用对HCT116细胞增殖的影响;采用Hoechst染色法和Annexin V/PI双染法检测细胞凋亡;JC-1染...

Effects of asiatic acid combined with oxaliplatin on the apoptosis,autophagy and pyroptosis of colon cancer HCT116 cells

OBJECTIVE To investigate the regulatory effects of asiatic acid(AA) combined with oxaliplatin(OXP) on the proliferation, apoptosis, autophagy and pyroptosis of colon cancer HCT116 cells.METHODS The effects of AA, OXP and the combination of them on HCT116 cell proliferation were detected by CCK8 assay. Apoptosis was detected by Hoechst staining and Annexin V/PI double staining. Mitochondrial membrane potential was detected by JC-1 staining. Cleaved Caspase-3, Bax, Bcl-2, GSDME, Beclin-1, LC3 and p62 were detected by Western blot.RESULTS AA and OXP inhibited the proliferation of colon cancer HCT116 cells, and the IC₅₀ values after 24 h were 40.80 μmol·L^-1 and 40.64 μmol·L^-1, respectively. AA significantly enhanced the inhibition of HCT116 cell proliferation of OXP compared with OXP alone(P<0.01). AA also significantly enhanced the apoptotic effect of OXP(P<0.01). AA could promote the change of mitochondrial membrane potential induced by OXP. AA significantly enhanced such abilities of OXP as down-regulating the expression of p62(P<0.01), and up-regulating the expression of Cleaved Caspase-3, GSDME and Beclin-1(P<0.01) as well as the ratios of LC3-Ⅱ/LC3-Ⅰ and Bax/Bcl-2(P<0.01).CONCLUSION AA can synergically increase the effects of OXP on the proliferation inhibition, apoptosis, autophagy and pyroptosis of HCT116 cells, and the synergistic mechanism might be related to the down-regulation of p62 expression, and up-regulation of Cleaved Caspase-3, GSDME and Beclin-1 expression as well as the ratios of LC3-Ⅱ/LC3-Ⅰ and Bax/Bcl-2. The results of this study provide experimental basis for the clinical application of AA combined with OXP in anti-tumor treatment.