Experimental Study On Apoptosis-promoting Effect Of BNIP-3Induced By HIF-1in A172Human Glioblastoma Cell Lines

Objective The purpose of this research was to detect the expression of HIF-1and BNIP-3in A172human glioblastoma cells under hypoxia environment and find the relationshipsbetween them,and to investigate the roles of inhibiting proliferation and inducing ofapoptosis to glioblastoma cells.Methods1. We cultured A172human glioblastoma cell lines and created hypoxia boxmodel,then randomly divided the cells into four groups.They are group A (control group),group B (hypoxia group), group C (hypoxia+HIF’s agonist) and groupD(hypoxia+HIF’s agonist+BNIP-3’s inhibitor). The cells of each group were observed byphase contrast microscope at24h,48h,72h time points to learn about morphologicalchanges.The inhibition of cell proliferation in each group was detected with MTT method.2. Apoptosis and proliferation of A172cells was detected with Tunel-fluorescence method.3. The expression of protein HIF-1and BNIP-3in each group was detected usingimmunohistochemistry.Results1.Phase contrast microscope detection: Compared with group A, cell survivals ofgroup B,C and D were significantly reduced and proliferation was inhibited which wasmore significant in group B and C. MTT method detection:cell proliferation of group B,Cand D was inhibited which was more significant in group B and C.2.Tunel fluorescencedetection: Compared with group A, cell survival and proliferation of group B and C werestatistically significant (p <0.05); Compared with group B, cell proliferation and survivalof group C were statistically significant(p<0.05). Compared with group C, cellproliferation and survival of group D were statistically significant(p<0.05).3Immunohistochemical detection:(1) Protein HIF-1expression: Compared with group A,HIF-1expression of group B,C and D was significantly increased,and the difference wasstatistically significant (p <0.05); Expression of HIF-1in group C and D was increasedcompared with group B,and the difference was statistically significant (p <0.05).(2)Protein BNIP-3expression: BNIP-3expression of group D was significantly reducedcompared with that of group B and C.BNIP-3expression of the other three groups at eachtime point was positively correlated with protein HIF-1expression which had statisticalsignificance. Conclusion1.A172human glioma cell proliferation could be inhibited under hypoxicenvironment, and cell apoptosis was caused.2.HIF-1expression increased underhypoxic environment, and could induce BNIP-3generation, which were positivelycorrelated.3.HIF-1could induce apoptosis of A172cells,and the possible mechanismmight be related to the BNIP-3generation induced by HIF-1.