miR-497对前列腺癌细胞生长侵袭及奥沙利铂药物敏感性的影响

Effects of miR-497 on the growth and invasion of prostate cancer cells and the sensitivity of oxaliplatin

目的探讨miR-497对人前列腺癌细胞系PC3-AR生长侵袭及奥沙利铂药物敏感性。方法用RT-PCR检测正常前列腺上皮细胞系RWPE-1和人前列腺癌细胞系PC3-AR中miRNA-497的表达水平。将PC3-AR分别转染不同浓度miR-497模拟物(25、50和100 nmol/L),通过噻唑蓝法和Transwell法检测miR-497对PC3-AR细胞增殖、迁移和侵袭能力的影响。用双报告基因检测miR-497与IκB激酶β(IKKβ)mRNA的3′-UTR的结合。用免疫印迹分析miR-497对抗基质金属蛋白酶9(MMP-9)、细胞周期蛋白依赖性蛋白激酶8(CDK8)、IKKβ、前列腺特异性抗原(PSA)的影响。分析miR-497对PC3-AR细胞对奥沙利铂敏感性影响。结果miRNA-497在PC3-AR中的表达水平显著下调(P<0.01)。过表达的miR-497显著抑制PC3-AR的增殖、迁移和侵袭。miR-497通过抑制IKKβ的蛋白表达,进一步抑制CDK8、MMP-9和PSA的蛋白表达,从而抑制PC3-AR的增殖和迁移。miR-497通过抑制奥沙利铂处理的PC3-AR细胞增殖并促进其凋亡,增加PC3-AR细胞对奥沙利铂的敏感性。结论miR-497通过调节PC3-AR细胞的IKK-β而起到肿瘤抑制基因的作用,miR-497可增加PC3-AR细胞对奥沙利铂药物的敏感性。

Objective To investigate the effect of miR-497 on the growth and invasion of human prostate cancer cell line PC3-AR and oxaliplatin.Methods RT-PCR was used to detect the expression level of miRNA-497 in normal prostate epithelial cell line RWPE-1 and human prostate cancer cell line PC3-AR.The logarithmic growth phase PC3-AR was transfected with different concentrations of miR-497 mimics(25,50 and 100 nmol/L)respectively,and miR-497 was detected by MTT assay and Transwell assay were used to assay the effects of proliferation,migration and invasiveness of PC3-AR.The dual reporter gene was used to detect the binding of miR-497 to the 3′-UTR of IKKβmRNA.The effect of miR-497 on the protein expression of IKKβ,CDK8,MMP9 and PSA was analyzed by Western blot.The effect of miR-497 on the sensitivity of PC3-AR cells to oxaliplatin was investigated.Results The expression level of miRNA-497 in PC3-AR was significantly down-regulated(P<0.01).Overexpressed miR-497 can significantly inhibite the proliferation,migration and invasion of PC3-AR.miR-497 can inhibit the protein expression of CDK8,MMP-9 and PSA by inhibiting the protein expression of IKKβ,thereby inhibiting the proliferation and migration of PC3-AR.miR-497 can increase the sensitivity of PC3-AR cells to oxaliplatin by inhibiting the proliferation of oxaliplatin-treated PC3-AR cells and promoting their apoptosis.Conclusions miR-497 acts as a tumor suppressor gene by modulating IKK-βin PC3-AR cells,and miR-497 increases the sensitivity of PC3-AR cells to oxaliplatin.