Influence Of Knocking Down HIF-1α Gene With SiRNA On The Biological Behavior Of Human Prostate Cancer PC3 Cell

Objective To investigate influence of knocking down HIF-1αgene with small interference RNA(siRNA) on biological behavior of human prostate cancer PC3 cell, including cell proliferation, migration force, invasiveness, apoptosis, expression of VEGF and GLUT-1, et al.Methods PC3 cell were transfected with HIF-1αsiRNA and NC siRNA confirmed by the preceding study using cationic liposomes Lipofectamine 2000. PC3 cell was divided into three groups: PC3 group, PC3+ NC siRNA group and PC3 + HIF-lαsiRNA group; CCK-8 test was used to measure the proliferation of PC3 cell; Scarification test was used to observe migration; Transwell test was used to observe invasiveness; Flow cytometer was used to measure cell apoptosis; ELISA test was used to detect protein expression of VEGF and GLUT-1.Results①.Cell proliferation was estimated at 0h, 12h, 24h, 48h, 72h and 96h after transfection by CCK-8 test. Comparing the survival rate (according to the measured OD values of cells) of three groups of cells, We found ,at 24h, 48h, 72h and 96h after transfection, survival rate of PC3+HIF-lαsiRNA group was different to the others(P<0.05), and the interference effect in 48h was the most efficient of all(F=53.77,P<0.01).②. Scarification test was used to observe migration capacity after transfection, Our results demonstrated that cell migrate of PC3 + HIFl-αsiRNA group was slow; while cell migrate of PC3 group was faster. The difference between PC3 + HIF-lαsiRNA group and PC3 group was great(F=134.02,P<0.01), and the difference between PC3 +NC siRNA group and PC3 group was not obviously(P>0.05);③.Transwell method was used to measure the invasiveness after transfection. Results showed that: the cell number of PC3 + HIF-lα siRNA group going through the Matrigel gel were significantly less than the number of PC3 group and PC3 + NC siRNA group (F=28.00,P<0.01), while the cell number between PC3 group and PC3 + NC siRNA group was no large difference (P>0.05);④.Flow cytometer was used to measure cell apoptosis at 48h after transfection. Results showed that: the cell apoptosis rates of PC3 + HIF-lαsiRNA group, PC3 group and PC3 + NC siRNA group was (11.2±1.13)%, (2.4±0.26)% and(2.5±0.36)%, respectively. The apoptosis of PC3 +HIF-lαsiRNA group was high(F=150.14,P<0.01), while the cell apoptosis rate of PC3 group and PC3 + NC siRNA group was no large difference (P>0.05);⑤.ELISA was used to detect protein expression of VEGF and GLUT-1 of three groups at 48h after transfection. Comparing its OD value, the results showed that: the VEGF protein and GLUT-1 protein levels of PC3+HIF-lαsiRNA group decreased significantly than the other groups(FVEGF=277.50,PVEGF<0.01;F GLUT-1=691.41,P GLUT-1<0.01), while the PC3 group and PC3 + NC siRNA group showed similar(PVEGF>0.05;P GLUT-1>0.05);Conclusion①.Knocking down HIF-1αgene can effectively inhibit the growth in PC3 cells;②.Knocking down HIF-1αgene can effectively decrease the migration and invasion capacity in PC3 cells;③.Knocking down HIF-1αgene may induce the apoptosis in PC3 cells;④.Knocking down HIF-1αgene can downregulate expression of VEGF and GLUT-1;...