miR-363-3p表达异常对人前列腺癌细胞生物学表型的影响

Effects of miR-363-3p expression on biological behaviors of human prostate cancer cells

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 investigate the expression of miR-363-3p in human prostate cancer cells and normal prostate epithelial cells, and explore its impact on the biological behaviors of prostate cancer cells. Methods Total RNA in human prostate cancer cells DU145, PC3 and normal prostate epithelial cells RWPE-1 were extracted respectively. The expressions of miR-363-3p in prostate cancer cells and normal prostate epithelial cells were detected by SYBR Green real-time PCR（RT-PCR）. DU145 cells and PC3 cells were infected with the synthesized PCDH, miR-363-3p and miR-363-3p sponge, respectively. CCK 8 and Colony assay were utilized to evaluate cell proliferation, transwell assay was applied to detect migration ability, and microtubule assay to measure tube formation ability. Results The expressions of miR-363-3p in the DU-145 and PC3 cells were all significantly lower than that in the RWPE-1 cells （P〈0.05）. Compared with the control, overexpression of miR-363-3p can significantly enhance cell proliferation, migration and invasion, and microtubule formation of DU-145 and PC3 cells （P〈0.05）. In contrast, knockdown of miR-363-3p significantly reversed these changes in biological phenotypes mentioned above （P〈0.05）. Conclusion miR-363-3p was upregulated in prostate cancer cells. Overexpression of miR-363-3p can promote cell proliferation, invasion and migration and microtubule formation. miR-363-3p may play an important role in the development of prostate cancer, which may become a new target for treatment of prostate cancer.