微RNA-21靶向调控SOX7基因影响胃癌BGC-823细胞增殖、迁移和侵袭

Effect of miR-21 on proliferation,migration and invasion of gastric cancer BGC-823 cells by targeting SOX7 gene

目的探讨微RNA(miR)-21对胃癌BGC-823细胞增殖、迁移和侵袭及SOX7表达的影响。方法采用脂质体转染法将miR-21抑制物及其阴性对照分别转染BGC-823细胞,分别记为anti-miR-21组和anti-NC组,以未转染BGC-823细胞记为空白对照(Control组)。采用实时荧光定量PCR(qRT-PCR)检测各实验组miR-21水平,四甲基噻唑蓝(MTT)法比较各组细胞BGC-823细胞增殖情况,划痕和Transwell实验分别比较各组BGC-823细胞迁移和侵袭能力。免疫印迹法检测各组细胞中SOX7蛋白表达,双荧光素酶报告基因实验检测miR-21与SOX7靶向调控关系。结果qRT-PCR检测结果显示,转染48 h后anti-miR-21组BGC-823细胞中miR-21的表达水平明显低于anti-NC组和Control组(P<0.05)。MTT检测结果显示,anti-miR-21组BGC-823细胞增殖能力明显高于anti-NC组和Control组(P<0.05)。划痕实验结果显示,anti-miR-21组BGC-823细胞相对迁移距离明显大于anti-NC组和Control组(P<0.05)。Transwell实验结果显示,anti-miR-21组BGC-823细胞侵袭细胞数明显高于anti-NC组和Control组(P<0.05)。免疫印迹显示anti-miR-21组BGC-823细胞中SOX7表达水平明显高于anti-NC组和Control组(P<0.05)。双荧光素酶报告基因实验显示,miR-21可显著降低SOX7野生型转染细胞相对荧光素酶活性,对SOX7突变型转染细胞的相对荧光素酶活性无明显影响。结论miR-21靶向调控SOX7基因影响胃癌BGC-823细胞增殖、迁移和侵袭。

Objective To investigate the effects of miR-21 on proliferation,migration and invasion of gastric cancer BGC-823 cells and the expression of SOX7.Methods BGC-823 cells were transfected with miR-21 inhibitor(anti-miR-21 group)and its negative control(anti-NC group)by lipofection,respectively.Untransfected BGC-823 cells were usedasblank controls(control group).Real-time quantitative PCR(qRT-PCR)was used to detect the level of miR-21 in each experimental group.The proliferation of BGC-823 cells was measured by MTT assay and compared among the groups.Scratch and Transwell assay were used tocompare the migration and invasion abilityof BGC-823 cellsamong the groups.Western blotting was used to detect the expression of SOX7 protein in each group.Dual luciferase reporter gene assay was used to detect the targetingrelationship between miR-21 and SOX7.Results At 48 h after tranfection,qRT-PCR showed that the expression level of miR-21 in BGC-823 cells ofanti-miR-21 group was significantly lower than that of the anti-NC group and control group(P<0.05);MTT assay showed that the proliferation of BGC-823 cells in anti-miR-21 group was significantly more prominent than that in anti-NC group and control group(P<0.05);scratch test showed that the relative migration distance of BGC-823 cells in anti-miR-21 group was significantly greater than that in the anti-NC group and control group(P<0.05);Transwell assay showed that the number of invasive BGC-823 cells in theanti-miR-21 group was significantly higher than that in theanti-NC group and control group(P<0.05);Western blottingshowed thatthe expression level of SOX7 in BGC-823 cells ofanti-miR-21 group was significantly higher than that of anti-NC group and control group(P<0.05);dual luciferase reporter gene assayshowed that miR-21 significantly reduced the relative luciferase activity of SOX7 wild type transfected cellsbut had no significant effect on the relative luciferase activity of the SOX7 mutant transfected cells.Conclusion miR-21 affects the proliferation,migration and invasion of gastric cancer BGC-823 cells by targeting and regulating SOX7 gene.