微小RNA-448抑制剂对人前列腺癌细胞增殖和迁移的影响

摘要

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 fluo-rescent quantitative polymerase chain reaction ( qRT-PCR) was used to detect the expression of miR-448 in normal prostate epi-thelial cells and cancer cells. The cells with the highest expres-sion of miR-448 were selected for follow-up experiment. The miR-448 inhibitor or miR-NC was transferred into prostate canc-er 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. Tr-answell assay was used to detect the cell migration ability. Re-sults 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 in-hibitor (P＜0. 01). The expression of CMTM3 mRNA was up-regulated (P＜0. 01). The expression of CMTM3, E-cadherin and β-catenin proteins were up-regulated. The expression of N-cadherin and Snail proteins were down-regulated. Cell prolifera-tion 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 expres-sion of CMTM3 protein, inhibit the proliferation and migration of prostate cancer cells, which showing a potential function in mo-lecular therapy of prostate cancer.%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LNCaP)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.