miR-1271抑制卵巢癌细胞的增殖和转移的研究

miR-1271 inhibits the proliferation and metastasis of ovarian cancer cells

目的探讨miR-1271对卵巢癌细胞的增殖、侵袭和迁移的影响,及其作用机制。方法实验分为miR-NC组(转染miR-NC)、miR-1271组(转染miR-1271 mimics)、miR-1271+pcDNA3.1组(共转染miR-1271和pcDNA3.1)、miR-1271+pcDNA3.1-Twist1组(共转染miR-1271和pcDNA3.1-Twist1)。用噻唑蓝法检测细胞活性,用Transwell检测细胞迁移和侵袭情况,用实时定量聚合酶链反应检测miR-1271和碱性螺旋-环-螺旋转录因子1(Twist1)mRNA的表达水平,用荧光素酶报告基因检测实验检测miR-1271对Twist1的靶向调控。结果干预24 h后,miR-NC组、miR-1271组、miR-1271+pcDNA3.1组和miR-1271+pcDNA3.1-Twist1组的细胞活性分别为0.71±0.07,0.46±0.04,0.42±0.04和0.57±0.05,迁移细胞数分别为(129.00±9.69),(68.00±3.26),(67.00±3.18)和(98.00±6.85)个,侵袭细胞数分别为(110.00±8.16),(53.00±2.91),(55.00±2.96)和(83.00±4.39)个,miR-1271表达水平分别为0.18±0.02,0.47±0.04,0.46±0.06和0.49±0.01,Twist1 mRNA表达水平分别为0.68±0.06,0.13±0.01,0.11±0.01和0.47±0.03。miR-1271组和miR-1271+pcDNA3.1-Twist1组的细胞活性、迁移和侵袭细胞数、miR-1271和Twist1 mRNA表达水平分别与miR-NC组和miR-1271+pcDNA3.1组比较,差异均有统计学意义(均P<0.05)。转染WT-Twist1后,相较于miR-NC组,miR-1271组的荧光素酶活性显著降低(0.63±0.06 vs 1.12±0.10,P<0.05);而转染MUT-Twist1后,相较于miR-con组,miR-1271组的荧光素酶活性无显著变化(1.09±0.10 vs 1.11±0.10,P>0.05)。结论miR-1271可抑制卵巢癌细胞的增殖、迁移和侵袭,其机制可能与Twist1有关。

Objective To investigate the effects of miR-1271 on proliferation,invasion and migration of ovarian cancer cells and its mechanism.Methods The experiment was divided into miR-NC group(transfected miR-NC),miR-1271 group(transfected miR-1271 mimics),miR-1271+pcDNA3.1 group(co-transfection of miR-1271 and pcDNA3.1),and miR-1271+pcDNA3.1-Twist1 group(co-transfection of miR-1271 and pcDNA3.1-Twist1).Methyl thiazolyl tetrazolium was used to detect cell viability.Transwell was used to detect cell migration and invasion.Reverse transcription-polymerase chain reaction was used to detect the expression levels of miR-1271 and Twist1 mRNA.The luciferase reporter gene detection experiment was used to detect the targeted regulation of miR-1271 on Twist1.Results After 24 h of intervention,the cell viability of the miR-NC group,miR-1271 group,miR-1271+pcDNA3.1 group and miR-1271+pcDNA3.1-Twist1 group were 0.71±0.07,0.46±0.04,0.42±0.04 and 0.57±0.05,the numbers of migrating cells were 129.00±9.69,68.00±3.26,67.00±3.18 and 98.00±6.85,the numbers of invasive cells were(110.00±8.16),(53.00±2.91),(55.00±2.96)and(83.00±4.39),the expression levels of miR-1271 were 0.18±0.02,0.47±0.04,0.46±0.06,0.49±0.01,the expression levels of Twist1 mRNA were 0.68±0.06,0.13±0.01,0.11±0.01,0.47±0.03,respectively.The cell viability,numbers of migration and invasion cells,expression levels of miR-1271 and Twist1 mRNA in the miR-1271 group and miR-1271+pcDNA3.1-Twist1 group were compared with miR-NC group and miR-1271+pcDNA3.1 group,respectively,the differences were statistically significant(all P<0.05).After transfection with WT-Twist1,compared with miR-NC group,the luciferase activity of miR-1271 group was significantly reduced(0.63±0.06 vs 1.12±0.10,P<0.05);while after transfection with MUT-Twist1,compared with the miR-con group,the luciferase activity of the miR-1271 group did not change significantly(1.09±0.10 vs 1.11±0.10,P>0.05).Conclusion miR-1271 can inhibit the proliferation,migration and invasion of ovarian cancer cells,and its mechanism may be related to Twist1.