miR-193a靶向调控HOTAIR对前列腺癌细胞活力及侵袭能力的影响

Effect of miR-193a on viability and invasion of prostate cancer cells via HOTAIR

目的探讨miR-193a通过靶向调控长链非编码RNA-Hox基因的反义基因间RNA(HOTAIR)对前列腺癌细胞活力及侵袭能力的影响。方法体外培养正常前列腺上皮细胞RWPE-1、前列腺癌(Pca)细胞DU145、PC3细胞株,采用实时荧光定量PCR(qRT-PCR)检测三组细胞的miR-193a及HOTAIR表达量。以慢病毒为载体构建过表达miR-193a的DU145、PC3细胞株,并设立阴性对照组,采用qRT-PCR法检测转染后细胞的HOTAIR的表达水平,CCK-8法检测细胞活力,Transwell法检测细胞侵袭能力。结果与正常前腺上皮细胞株相比,DU145中miR-193a的相对表达量约为42%,PC3中miR-193a相对表达量约为27%,DU145中HOTAIR表达上调约2.6倍,PC3中HOTAIR表达上调约4.2倍,差异均有统计学意义(P<0.05);成功转染过表达mR-193a后,与阴性对照组比较,DU145中miR-193a表达量上q调2.4倍,PC3中miR-193a表达量上调3.1倍,DU145中HOTAIR相对表达量约为41%,PC3中HOTAIR相对表达量为30%,差异均有统计学意义(P<0.05)。DU145细胞株转染miR-193a后,细胞穿膜次数为(72.31±11.54),对照组细胞穿膜次数为(116.63±26.74),差异有统计学意义(P<0.05);PC3细胞株转染miR-193a后,细胞穿膜次数为(67.16±10.38),对照组细胞穿膜次数为(145.61±21.94),差异有统计学意义(P<0.01)。结论 miR-193a可以抑制体外前列腺癌细胞长链非编码基因HOTAIR的表达,降低细胞活力及侵袭能力,对于前列腺癌的治疗提供了新的思路。

Objective To investigate the effect of miR-193a on the viability and invasion ability of prostate cancer cells by targeting and regulating long-chain non-coding RNA-Hox transcript antisense intergenic RNA( HOTAIR). Methods The normal prostate epithelial cells RWPE-1 and prostate cancer(Pca) cells DU 145 and PC3 were cultured in vitro. The expression of miR-193a and HOTAIR were detected by real-time quantitative PCR(qRT-PCR). The lentivirus vector was used to construct DU 145 and PC3 cell lines overexpressing miR-193a, and a negative control group was set up. HOTAIR expression was detected by qRT-PCR. Cell viability was measured by CCK-8 and cell invasiveness was detected by transwell method. Results Compared with normal anterior glandular epithelial cell line, the relative expression of miR-193a in DU 145 was about 42%;the relative expression of PC3 was about 27%;the expression of HOTAIR in DU 145 was upregulated by about 2.6-fold, and the expression of PC3 was up-regulated by about 4.2-fold. After successful transfection of mR-193 a, the expression of miR-193a was up-regulated by 2.4-fold;the expression of PC3 was up-regulated by 3.1-fold,and the relative expression of HOTAIR was about 41% in DU145. The relative expression of PC3 was compared with the negative control group. The cell viability of DU 145 and PC3 was significantly decreased compared with the control group.After transfection of miR-193a,the number of transmembrane cells was( 72.31 ± 11.54),and the number of permeabilization of NC group was( 116.63 ±26.74). There was a statistically significant difference between the two group(P<0.05). After transfection of miR-193a in PC3 cell line, the number of transmembrane cells was(67.16± 10.38),and the number of transmembrane in NC group was( 145.61 ±21.94),and the difference was significant(P<0.01). Conclusion miR-193a could inhibit the expression of long-chain non-coding gene HOTAIR in vitro and reduce the cell viability and invasion ability,which provides a new idea for the treatment of prostate cancer.