稳定过表达microRNA―96前列腺癌细胞株的建立及其对前列腺癌细胞增殖和迁移的影响.docVIP

稳定过表达microRNA―96前列腺癌细胞株的建立及其对前列腺癌细胞增殖和迁移的影响 　　[摘要] 目的 构建稳定过表达microRNA-96（miR-96）的前列腺癌DU-145细胞株并研究其对DU-145细胞增殖和迁移的影响。 方法 构建miR-96慢病毒表达载体并转染前列腺癌细胞DU-145，嘌呤霉素筛选出稳定表达miR-96（实验组）及阴性对照（对照组）的细胞株；通过实时荧光定量PCR（qRT-PCR）检测转染后两组细胞miR-96的表达；克隆形成实验和MTT实验检测两组的细胞增殖情况。transwell迁移实验检测两组细胞的迁移能力。 结果 经过筛选后各组转染细胞发光效率均 90%；qRT-PCR结果显示实验组miR-96表达水平明显高于对照组（P 0.05）；克隆形成实验显示实验组克隆形成率明显低于对照组（P 0.05）；MTT实验显示实验组各个时间点吸光度值均低于对照组（P 0.05）；transwell迁移实验表明实验组穿膜细胞数明显少于对照组（P 0.05）。 结论 miR-96可抑制前列腺细胞的增殖和迁移能力，可能参与前列腺癌的发生、发展。 　　[关键词] 前列腺癌；microRNA-96；增殖；迁移 　　[中图分类号] R737.25 [文献标识码] A [文章编号] 1673-7210（2016）01（b）-0024-05 　　[Abstract] Objective To establish microRNA-96 （miR-96） stable overexpression cell lines of cervical cancer and detect its effect on proliferation of prostatic cancer cell lines DU-145. Methods Lentiviral vector stable overexpressing miR-96 was constructed and prostatic cancer cell lines DU-145 were transfected （experimental group）. The transfection cells were screened by using of puromycin and the stable transfection cell lines were obtained （control group）. Endogenous miR-96 levels in two groups were measured by quantitative RT-PCR（qRT-PCR）. The proliferation of DU-145 cells in two groups were detected by clonogenic proliferation assay and MTT assay. The migration of DU-145 cells in two groups were assessed by transwell assay. Results After screening， luminous efficiency rate of transfection cell in each group was over 90%. qRT-PCR showed that the expression level of miR-96 in experimental group was significantly higher than that in control group （P 0.05）. Colony formation assay showed that the clonogenicity rate of experimental group was significantly lower than that of control group （P 0.05）. MTT assay demonstrated that absorbance values all time points of experimental group were lower than those of control group （P 0.05）. Transwell assay revealed that the number of cells through the well in experimental group were less than that in control group （P 0.05）. Conclusion miR-9