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目的研究微小RNA-363-3p(miR-363-3p)在人前列腺癌细胞和正常前列腺上皮中的表达并探究miR-363-3p对前列腺癌细胞生物学表型的影响。方法分别提取人前列腺癌细胞DU145、PC3和正常肝细胞RWPE-1的总RNA,采用实时荧光定量PCR(RT-PCR)法检测miR-363-3p的表达。运用脂质体介导的方法分别将慢病毒空载体pCDH-vector(简称PCDH)及构建好的慢病毒质粒pCDH-miR-363-3p(简称miR-363-3p质粒)和pCDH-miR-363-3p sponge(miR-363-3p sponge质粒)转染DU145、PC3细胞,利用CCK8和平板克隆实验检测miR-363-3p表达改变后对细胞增殖能力的影响;Transwell实验检测miR-363-3p表达变化对细胞体外迁移侵袭能力的影响;微管实验检测miR-363-3p表达变化对细胞体外成管能力的改变。结果 miR-363-3p在人前列腺癌细胞与正常前列腺上皮细胞中的表达水平相比,显著上调(P<0.05)。体外实验中,过表达miR-363-3p能显著增强人前列腺癌细胞的增殖、迁移侵袭及微管形成能力,下调miR-363-3p后上述生物学表型均受到抑制,且差异显著。结论miR-363-3p在人前列腺癌细胞中表达上调,过表达miR-363-3p可促进人前列腺癌细胞体外增殖、迁移侵袭和微管形成能力。在前列腺癌发病进程中,miR-363-3p可能扮演促癌基因的角色,有望成为前列腺癌治疗的新靶点。
Objective To study the expression of miR-363-3p (miR-363-3p) in human prostate cancer cells and normal prostate epithelium and to explore the effect of miR-363-3p on the biological phenotypes of prostate cancer cells. Methods The total RNA of human prostate cancer cells DU145, PC3 and normal liver cells RWPE-1 were extracted and the expression of miR-363-3p was detected by real-time fluorescence quantitative PCR (RT-PCR). The lentiviral empty vector pCDH-vector (PCDH) and the constructed lentiviral plasmid pCDH-miR-363-3p (referred to as miR-363-3p plasmid) and pCDH-miR-363 -3p sponge (miR-363-3p sponge plasmid) was transfected into DU145 and PC3 cells. The effect of miR-363-3p expression on cell proliferation was detected by CCK8 and plate cloning experiments. Transwell assay was used to detect miR-363-3p expression The effect of changes of miR-363-3p expression on cell migration and invasion in vitro was detected by microtubule assay. Results miR-363-3p was significantly up-regulated in human prostate cancer cells compared with normal prostate epithelial cells (P <0.05). In vitro, overexpression of miR-363-3p significantly enhanced the proliferation, migration, invasion and microtubule formation ability of human prostate cancer cells. The down-regulation of miR-363-3p suppressed the above biological phenotypes and showed significant differences. Conclusion miR-363-3p is upregulated in human prostate cancer cells. Overexpression of miR-363-3p can promote proliferation, migration, invasion and microtubule formation of human prostate cancer cells in vitro. In the process of prostate cancer, miR-363-3p may play a role of oncogene, which is expected to become a new target for the treatment of prostate cancer.