The Effect And Mechanism Of Free Fatty Acids In Biology Behavior Of Prostate Cancer Cell

Object: Cult ivation of human prostate cancer cells PC-3 in vitro,the research intends to observe the effects of free fatty acid(FFA)in biological behaviour of PC-3 cells,and try to explore the molecular mechanisms possiblely.Methods:Cultivation of human prostate cancer cells PC-3 in vitro with FFA,which mixed by oleic acid(OA)and palmitic acid(PA)on 2:1.The m RNA expression and protein expression of FABP4,PPAR?,VEGF-A,m TOR,Vimentin and AMP K was detected by real-time fluorescent quantitative PCR,western blot and immunohistochemical technology.Cell cycle was detected by flow cytometry instrument.Cell invasion and migration was detected by transwell chamber experiment.Using cell transfection and si-RNA method to up or down expression of PPAR?,and si-RNA method to down expression of PPAR? treat with FFA at the same time.And then the m RNA expression of VEGF-A,m TOR and Vimentin was detected by real-time fluorescent quantitative PCR,the glucolipid metabolism ability was detected by kit,the cell cycle was detected by flow cytometry instrument,the cell invasion and migration was detected by transwell chamber experiment.Using cell transfection and si-RNA method to up or down expression of AMPK,then detecte the biological behaviour of PC-3 cells.Results: 1.1.5 m M FFA treat on the PC-3 cells after 48 hours,the m RNA expression and protein expression level of FABP4,PPAR? and VEGF-A in FFA group were significantly higher than those in the blank group(P<0.05).The m RNA expression level of m TOR,Vimentin,AMP K in FFA group were signif icantly higher than those in the blank group(P<0.05).The amount of PC-3 cells in G0/G1 phase were signif icantly lower,while S phase in FFA group were signif icantly higher than those in the blank group(P<0.05).The ablity of cell m igration and invasion in FFA group were significantly higher than those in the blank group(P<0.05).2.Up-regulated PPAR? after 48 hours,the m RNA expression level of VEGF-A,m TOR and Vimentin in Ad-PPAR? group were significantly higher than those in the mock group(P<0.05).The protein expression level of VEGF-A in Ad-PPAR? group were significantly higher than those in the mock group(P<0.05).The lipid content of PC-3 in Ad-PPAR? group were signif icantly higher than those in the mock group(P<0.05).The amount of PC-3 cells in G0/G1 phase in Ad-PPAR? group were signif icantly lower,while S phase and G2/M phase in Ad-PPAR? group were significantly higher than those in the mock group(P<0.05).The ablity of cell migration and invasion in Ad-PPAR? group were signif icantly higher than those in the mock group(P<0.05).3.Down-regulated PPAR? after 24 hours,the m RNA expression level of VEGF-A,m TOR and Vimentin in si-PPAR? group were significantly lower than those in the NC group(P<0.05).The lipid content of PC-3 in si-PPAR? group were significantly lower than those in the NC group(P<0.05).The amount of PC-3 cells in G0/G1 phase in si-PPAR? group were signif icantly higher,while S phase and G2/M phase in si-PPAR? group were significantly lower than those in the NC group(P<0.05).The ablity of cell migration and invas ion in si-PPAR? group were significantly lower than those in the NC group(P<0.05).4.Down-regulated PPAR? and treat with FFA at the same time,the m RNA expression level of VEGF-A and Vimentin in si-PPAR?+FFA group were significantly lower than those in the FFA group(P<0.05).The amount of PC-3 cells in G0/G1 phase were significantly higher,while S phase in si-PPAR?+FFA group were signif icantly lower than those in the FFA group(P<0.05).The ablity of cell migration and invasion in si-PPAR?+FFA group were significantly lower than those in the FFA group(P<0.05).5.Up-regulated AMP K after 48 hours,the m RNA expression level of Vimentin in Ad-PPAR? group were significantly higher than those in the mock group(P<0.05).The ablity of cell proliferation,migration and invas ion in Ad-AMP K group were significantly higher than those in the mock group(P<0.05).Down-regulated AMP K after 24 hours,the m RNA expression level of Vimentin in si-AMPK group were significantly lower than those in the NC group(P<0.05).The ablity of cell proliferation,migration and invasion in si-AMP K group were signif icantly lower than those in the NC group(P<0.05).Down-regulated AMP K and treat with FFA at the same time,the ablity of cell proliferation,migration and invasion in si-AMP K+FFA group were significantly lower than those in the FFA group(P<0.05).Conclusions: 1.FFA promote the expression of VEGF-A,m TOR and Vimentin by up-regulate FABP4 and PPAR?,and promote the proliferation,migration and invas ion of human prostate cancer cells PC-3.2.FFA promote the expression of Vimentin by up-regulate AMPK,and promote the proliferation,migration and invasion of human prostate cancer cells PC-3.