微小 RNA-15a-5p靶向蛋白质二硫键异构酶 A6前体蛋白抑制前列腺癌细胞增殖、迁移及侵袭

目的 探讨微小RNA-15a-5p(miR-15a-5p)是否可通过靶向蛋白质二硫键异构酶A6前体蛋白(PDIA6)而抑制前列腺癌细胞增殖、迁移及侵袭.方法 选取2018年2月至2019年3月江南大学附属医院接受治疗的前列腺癌病人50例,采用实时荧光定量PCR(qRT-PCR)与蛋白印迹法(Western blotting)分别检测前列腺癌组织、癌旁组织中miR-15a-5p、PDIA6的表达量;体外培养人前列腺癌DU145细胞,将细胞分为miR-NC组、miR-15a-5p组、si-NC组、si-PDIA6组、miR-15a-5p+pcDNA组、miR-15a-5p+pcDNA-PDIA6组;噻唑蓝(MTT)检测细胞增殖;Transwell小室实验检测细胞迁移及侵袭;双荧光素酶报告实验验证miR-15a-5p、PDIA6的靶向关系.结果 与癌旁组织相比,前列腺癌组织中miR-15a-5p的表达水平降低(1.00±0.17)比(0.68±0.11)],PDIA6 mRNA(0.99±0.09)比(1.64±0.18)]和蛋白(0.44±0.07)比(0.76±0.17)]表达水平升高;转染miR-15a-5p mimics或转染si-PDIA6可降低细胞存活率(P<0.05),减少迁移及侵袭细胞数(P<0.05);双荧光素酶报告实验证实miR-15a-5p可靶向结合PDIA6;PDIA6过表达可降低miR-15a-5p过表达对DU145细胞增殖、迁移及侵袭的抑制作用.结论 过表达miR-15a-5p通过降低PDIA6的表达对列腺癌细胞增殖、迁移及侵袭能力有抑制作用.

miR-15a-5p inhibits prostate cancer cell proliferation, migration and invasion by targeting PDIA6

Objective To investigate whether miR-15a-5p could inhibit the proliferation, migration and invasion of prostate cancer cells by targeting PDIA6.Methods qRT-PCR and Western blotting were used to detect the expression of miR-15a-5p and PDIA6 inprostate cancer tissues and adjacent tissues. Human prostate cancer DU145 cells were cultured in vitro and divided into miR-NC group, miR-15a-5p group, si-NC group, si-PDIA6 group, miR-15a-5p+pcDNA group, miR-15a-5p+pcDNA -PDIA6 group. MTT wasused to detect the cell proliferation. The Transwell cell test was used to detect the cell migration and invasion. The dual luciferase report experiment verified the targeting relationship between miR-15a-5p and PDIA6.Results Compared with adjacent tissues, the expression level of miR-15a-5p in prostate cancer tissue decreased (1.00±0.17) vs. (0.68±0.11)], and the expression level of PDIA6 mRNA (0.99±0.09) vs. (1.64±0.18)] and protein (0.44±0.07) vs. (0.76±0.17)] increased. Transfection of miR-15a-5p mimics or transfection of si-PDIA6 could reduce cell survival rate (P<0.05), and reduce the number of migrating and invading cells (P<0.05). The dual luciferase report experiment confirmed that miR-15a-5p could target PDIA6. PDIA6 overexpression could reduce the inhibitory effect of miR-15a-5p overexpression on DU145 cell proliferation, migration and invasion.Conclusion Overexpression of miR-15a-5p inhibites the proliferation, migration and invasion of prostate cancer cells by reducing the expression of pdia6.