miR-409-3p通过靶向脂肪酸结合蛋白4影响子宫内膜癌细胞转移的研究

miR-409-3p Affects the Metastasis of Endometrial Cancer Cells by Targeting Fatty Acid-Binding Protein4

目的用生物信息学方法预测脂肪酸结合蛋白4(FABP4)的上游调控miRNA,体外培养子宫内膜癌细胞Ishikawa,了解微小核糖核酸-409-3p(miR-409-3p)是否通过抑制FABP4的表达水平,进而影响Ishikawa细胞的生物学行为。方法实验分为control组、miR-409-3p mimics组、miR-409-3p inhibitor组、miR-409-3p NC组、FABP4-OE组及FABP4-shRNA组6组,采用实时定量聚合酶链式反应及蛋白质印迹法检测miR-409-3p及FABP4在Ishikawa中的表达情况;采用细胞增殖实验(CCK-8)、细胞侵袭及迁移实验(Transwell法)检测miR-409-3p与FABP4对Ishikawa细胞增殖、侵袭以及迁移能力的影响;用双荧光素酶报告基因实验在Ishikawa细胞中检测miR-409-3p对FABP4的调控作用。结果1)在Ishikawa细胞中下调FABP4的表达后,Ishikawa细胞的增殖、侵袭和迁移能力均显著降低(P<0.05);在Ishikawa细胞中上调FABP4的表达后,Ishikawa细胞的增殖、侵袭和迁移能力均显著升高(P<0.05)。2)在Ishikawa细胞中转染miR-409-3p mimics后,FABP4的表达水平以及Ishikawa细胞的增殖、侵袭和迁移能力均显著降低(P<0.05);而转染miR-409-3p inhibitor后,FABP4的表达水平以及Ishikawa细胞的增殖、侵袭和迁移能力均显著升高(P<0.05)。3)双荧光素酶报告基因结果显示,在Ishikawa细胞中miR-409-3p可靶向调控FABP4。结论高表达的FABP4促进Ishikawa细胞增殖、侵袭及迁移,miR-409-3p可通过下调FABP4的表达进而抑制Ishikawa细胞发生转移。

Objective To predict the upstream regulatory miRNA of fatty acid-binding protein 4(FABP4)by bioinformatics methods,culture endometrial cancer Ishikawa cells in vitro,and determine whether microRNA(miR)-409-3p affects the biological behavior of Ishikawa cells by inhibiting the expression of FABP4.Methods There were 6 groups in the experiment:control group,miR-409-3p mimic group,miR-409-3p inhibitor group,miR-409-3p NC group,FABP4-OE group and FABP4-shRNA group.The expression of miR-409-3p and FABP4 in Ishikawa cells was detected by RT-qPCR and Western blot.CCK-8 and Transwell methods were used to detect the effects of miR-409-3p and FABP4 on the proliferation,invasion and migration ability of Ishikawa cells.The regulatory relationship between miR-409-3p and FABP4 was detected in Ishikawa cells by using double luciferase gene experiment.Results 1)After down regulating FABP4 in Ishikawa cells,the proliferation,invasion and migration ability of Ishikawa cells were significantly decreased(P<0.05).After up regulating FABP4 in Ishikawa cells,the proliferation,invasion and migration ability of Ishikawa cells were significantly increased(P<0.05).2)After transfection of miR-409-3p mimics into Ishikawa cells,the expression of FABP4 and the proliferation,invasion and migration ability of Ishikawa cells were significantly decreased(P<0.05).After transfection of miR-409-3p inhibitor into Ishikawa cells,the expression level of FABP4 and the ability of cell proliferation,invasion and migration were significantly increased(P<0.05).3)Dual luciferase reporter gene method showed that miR-409-3p could target FABP4 in endometrial cancer Ishikawa cells.Conclusion High expression of FABP4 promotes the proliferation,invasion and migration ability of Ishikawa cells.miR-409-3p may inhibit the metastasis of Ishikawa cells by down regulating the expression of FABP4.