Anti-apoptosis of hepatocyte growth factor on CML K562 cells / 中国药学杂志

ABSTRACT

OBJECTIVE: To investigate the protection of hepatocyte growth factor (HGF) on CML cell line K562 from apoptosis induced by etoposide (VP-16) and its molecular mechanism. METHODS: Quantitative and qualitative analyses on cell morphological change of apoptosis were performed through acridine orange (AO) staining and HE staining, and fluorescent flow cytometry.The test analyzes membrane on the surface of the PS evagination and integrity of cell membrane surface and mitochondrial membrane potential changes were performed through Annexin V-FITC/PI double dyeing and JC-1 cell dyeing tests, and apoptotic factors such as Bcl-2, Bax, Caspase-3 and Caspase-9 were measured by SYBR Green (Takara) qRT-PCR. RESULTS: The HE and AO staining revealed that apoptotic rates in HGF+VP-16 groups were significantly lower than those in VP-16 groups (P<0.05, P<0.05), HGF can inhibit the apoptosis of cells induced by VP-16; FCM (Annexin V-FITC/ PI and JC-1) tests showed that cells apoptotic rates in HGF+VP-16 groups were significantly lower than those in VP-16 groups (P<0.05, P<0.001), indicating that HGF has the anti-apoptosis function. Apoptosis related gene mRNA expression tests found that the Bcl-2 mRNA expression in HGF+VP group was obviously higher than that in the VP-16 group (P<0.001), while Bax mRNA, Caspase-3 mRNA, and Caspase-9 mRNA expressions were significantly lower than those in the VP-16 group (P<0.05, P<0.001, P<0.001), suggesting that HGF possesses antiapoptotic effect through inhibiting apoptosis gene expression and promoting the antiapoptotic gene expression simultaneously. CONCLUSION: HGF can significantly protect K562 cells from apoptosis induced by VP-16 through the HGF/c-Met way to regulate PI3K/AKT pathway.