微小RNA-221通过靶基因细胞因子信号抑制因子1表达水平调控前列腺癌细胞的侵袭力

Regulation of prostate cancer cells invasiveness via microRNA -221 by the expression level of target gene suppressor of cytokine signaling 1

目的观察前列腺癌细胞中微小RNA（miRNA，miR）-221的表达及其对前列腺癌细胞侵袭力的影响。方法应用实时定量反转录聚合酶链反应（RT-qPCR）检测miR-221在前列腺正常细胞株与前列腺癌细胞株中表达的差异，利用细胞转染构建miR-221过表达和抑制细胞株，再通过细胞迁移实验检测细胞侵袭力的变化；使用荧光素酶报告实验验证miR-221与细胞因子信号抑制因子1（SOCS1）基因的直接调控关系，再采用Western blot检测细胞中SOCS1基因表达水平的变化。结果RT-qPCR检测细胞株可见miR-221在PC3、LNCaP和DU145 3种前列腺癌细胞株中表达量（6.51、7.52和8.03）均比前列腺正常细胞株表达量（12.12）低（F＝235.852，P＝0.000），其中两两比较差异也有统计学意义。细胞迁移实验结果显示，过表达miR-221的前列腺肿瘤细胞迁移速度显著慢于对照组（25.3 h比17.8 h）。荧光素酶报告实验显示，miR-221能明显抑制SOCS1-3’UTR的荧光素酶活性（P＝0.006）。过表达miR-221后，PC3细胞中SOCS1的蛋白表达下调（P＝0.035）。结论miR-221负调控SOCS1能抑制前列腺癌细胞的迁移。

ObjectiveTo investigate microRNA （miRNA, miR）-221 expression in prostate cancer cells and its influence on prostate cancer cell invasiveness.MethodsmiR-221 expression levels in prostate cancer cell lines were measured by real-time quantitative reverse transcriptase-polymerase chain reaction （RT-qPCR）. The miR-221 overexpression and silencing cell lines were constructed by cell transfection. Effects of the depletion on cell invasiveness were assessed in vitro with Transwell. ResultsRT-qPCR showed miR-221 was down-regulated in PC3, LNCaP and DU145 cells than in PrEC （F=235.852, P=0.000）, in which pairwise comparison also had significant differences. Cell invasiveness assay showed that migration of LNCaP （P=0.000） and DU145 （P=0.000） cells overexpressing miR-221 was significantly slower than in the control group. ConclusionThe negative regulation of suppressor of cytokine signaling 1 （SOCS1） by miR-221 can inhibit the invasiveness of prostate cancer cells.