FFA上调AMPK/Vimentin促进前列腺癌细胞PC-3的增殖、侵袭和迁移

Up-regulation of AMPK/Vimentin by FFA promotes proliferation,invasion and migration of prostate cancer cell line PC-3

目的探讨游离脂肪酸(Free Fatty Acid,FFA)对人前列腺癌细胞PC-3生物学行为的影响及可能机制。方法1. 5mmol/L FFA混合液(油酸∶棕榈酸=2∶1)刺激体外培养的PC-3细胞,CCK-8法检测对细胞的毒性作用,流式细胞仪检测对增殖的影响,Transwell法检测对侵袭、迁移的影响。细胞转染和RNA干扰技术上/下调AMPK。qRT-PCR技术检测AMPK、Vimentin及VEGF的mRNA表达水平。Western Blot检测蛋白表达水平。结果 (1)1. 5 mmol/L FFA混合液处理48h后,PC-3细胞的增殖、侵袭和迁移能力均显著增强(P<0. 05),细胞内AMPK、Vimentin及VEGF的mRNA表达显著升高(P<0. 05)。(2) 1. 5 mmol/L FFA混合液作用于PC-3细胞的同时下调AMPK,PC-3细胞的增殖、侵袭和迁移能力显著降低(P<0. 05),Vimentin的mRNA表达显著降低(P<0. 05)。(3)上调AMPK表达48 h后,PC-3细胞的增殖、侵袭和迁移能力显著增强(P<0. 05),细胞内Vimentin的mRNA表达显著升高,VEGF的mRNA表达显著降低(P<0. 05);下调AMPK表达24 h后,PC-3细胞的增殖、侵袭和迁移能力均显著降低(P<0. 05),细胞内Vimentin的mRNA表达显著降低(P<0. 05),VEGF则无显著变化(P>0. 05)。结论高浓度FFA可能通过上调AMPK增加Vimentin表达,促进PC-3细胞的增殖、侵袭和迁移。

Objective To investigate the effect of free fatty acid( FFA) on the biological behavior of human prostate cancer cell line PC-3 and its possible mechanism.Methods 1. 5 mmol/L FFA mixture( oleic acid and palmitic acid mixed 2 ∶ 1) stimulated PC-3 cells cultured in vitro.The cytotoxicity of cells was detected by CCK-8 method,the effect on cell proliferation was detected by flow cytometry,and the effect on cell invasion and migration ability was detected by transwell method.Cell transfection and RNA interference technology to up-regulate and down-regulate AMPK.qRT-PCR technique was used to detect mRNA expression levels of AMPK,Vimentin and VEGF.Western Blot detect protein expression levels.Results( 1) After treatment with 1. 5 mmol/L FFA for 48 hours,the proliferation,invasion and migration of PC-3 cells were significantly increased( P<0. 05),and the mRNA expressions of AMPK,Vimentin and VEGF were significantly increased( P< 0. 05).( 2) Down-regulating of AMPK with 1. 5 mmol/L FFA intervention,the proliferation,invasion and migration ability of PC-3 cells were significantly decreased( P<0. 05),and the mRNA expression of Vimentin was also significantly decreased( P<0. 05).( 3) After 48 hours of up-regulation of AMPK expression,the proliferation,invasion and migration of PC-3 cells were significantly increased( P<0. 05),the mRNA expression of Vimentin was significantly increased,and the mRNA expression of VEGF was significantly decreased( P<0. 05);After 24 hours of AMPK expression down-regulation,the proliferation,invasion and migration of PC-3 cells were significantly decreased( P<0. 05),the mRNA expression of Vimentin was significantly decreased( P<0. 05),and the expression of VEGF was not significantly changed( P>0. 05).Conclusion High concentration of FFA may increase the expression of Vimentin by up-regulating AMPK,and promote the proliferation,invasion and migration of PC-3 cells.