微小RNA-448抑制剂对人前列腺癌细胞增殖和迁移的影响

Effect of microRNA-448 inhibitor on the proliferation and migration of human prostate cancer cells

目的观察特异性微小RNA-448抑制剂对前列腺癌细胞增殖和迁移的影响及其分子机制。方法 qRT-PCR检测正常前列腺上皮细胞(RWPE-1)和癌细胞株(DU-145、PC-3、C4-2B和LNCa P)中miR-448的表达,选取表达量的最高的癌细胞株进行后续实验。利用脂质体转染试剂将miR-448抑制剂或miR-NC转入前列腺癌细胞中,qRT-PCR检测miR-448及人趋化素样因子超家族成员3(CMTM3)mRNA的表达水平,Western blot检测细胞相关蛋白表达。MTT法检测细胞的增殖活性,Transwell实验检测细胞的迁移能力。结果 RWPE-1细胞中miR-448的表达显著低于癌细胞(P<0.01),DU-145细胞中的表达最高(P<0.01)。miR-448抑制剂转染48 h后,DU-145细胞miR-448表达下调(P<0.01),CMTM3 mRNA表达上调(P<0.01),CMTM3、E-cadherin和β-catenin蛋白表达上调,N-cadherin和Snail蛋白表达下调;细胞增殖活性降低(P<0.05);细胞迁移能力降低(P<0.01)。结论 miR-448在前列腺癌细胞中明显高表达,miR-448抑制剂可下调DU-145细胞中miR-448表达,上调CMTM3蛋白的表达,抑制前列腺癌细胞的增殖和迁移,在前列腺癌的分子治疗中具有潜在功能。

Purpose To observe the effect of specific miR-448 inhibitor on the proliferation and migration of prostate cancer cells and its molecular mechanism.Methods Real-time fluorescent quantitative polymerase chain reaction（qRT-PCR） was used to detect the expression of miR-448 in normal prostate epithelial cells and cancer cells.The cells with the highest expression of miR-448 were selected for follow-up experiment.The miR-448 inhibitor or miR-NC was transferred into prostate cancer cells using liposome transfection reagent.The expression of miR-448 and CMTM3 mRNA were detected by qRT-PCR.The expression of related proteins were detected by Western blotting.MTT assay was used to detect the cell proliferation activity.Transwell assay was used to detect the cell migration ability.Results The expression of miR-448 in normal prostate epithelial cells was significantly lower than that in cancer cells（P〈0.01）,and the expression of miR-448 was the highest in DU-145 cells（P〈0.01）.The expression of miR-448 in DU-145 cells was down-regulated 48 h after transfection with miR-448 inhibitor（P〈0.01）.The expression of CMTM3 mRNA was upregulated（P〈0.01）.The expression of CMTM3,E-cadherin and β-catenin proteins were up-regulated.The expression of Ncadherin and Snail proteins were down-regulated.Cell proliferation was decreased（P〈0.05）.Cell migration ability decreased（P〈0.01）.Conclusion miR-448 is highly expressed in prostate cancer cells.miR-448 inhibitor can down-regulate the expression of miR-448 in DU-145 cells,up-regulate the expression of CMTM3 protein,inhibit the proliferation and migration of prostate cancer cells,which showing a potential function in molecular therapy of prostate cancer.