miR-134靶向基质金属蛋白酶1对喉癌细胞的增殖、凋亡、侵袭和迁移的影响

Effect of miR-134 targeting MMP1 on proliferation,apoptosis,invasion and migration of laryngeal cancer cells

目的探讨miR-134靶向基质金属蛋白酶1(MMP1)对喉癌细胞的增殖、凋亡、侵袭和迁移的影响。方法双荧光素酶报告基因实验验证MMP1基因是否为miR-134的靶向基因。转染实验分为miR-134 mimics组、miR-134 antagomir组和空白对照组,采用CCK-8、流式细胞术和Transwell法检测各组细胞增殖、凋亡、侵袭和迁移情况,Western blot检测过表达或降低miR-134对MMP1蛋白表达的影响。结果双荧光素酶报告基因结果显示miR-134能和MMP13′-UTR端结合且明显抑制荧光素酶活性。miR-134 mimics组细胞在转染后24 h的细胞活力明显低于空白对照组(P<0.01),而miR-134 antagomir组明显高于空白对照组(P<0.05)。与空白对照组凋亡率[(4.32±0.36)%]比较,miR-134 mimics组[(12.02±0.45)%]明显增加(P<0.01),而miR-134 antagomir组[(2.31±0.26)%]明显降低(P<0.05)。与空白对照组比较,miR-134 mimics组的细胞侵袭和迁移能力明显减弱、MMP1蛋白表达明显降低(P<0.05),miR-134 antagomir组的细胞侵袭和迁移能力明显增强、MMP1蛋白表达明显升高(P<0.05)。结论miR-134在转录后水平调控MMP1蛋白表达来影响喉癌细胞的增殖、凋亡、侵袭及迁移。

Objective To determine the effects of miR-134 targeted matrix metalloproteinase 1(MMP1)on the proliferation,apoptosis,invasion and migration of laryngeal cancer cells.Methods The luciferase reporter gene assay verified whether MMP1 gene being miR-134 targeted gene.The transfection experiment was divided into the miR-134 mimics group,miR-134 antagomir group and blank control group.CCK-8,flow cytometry and Transwell assay were used to detect the proliferation,apoptosis,invasion and migration of the cells in each group.Western blot was used to detect the effect of over-expression or decreasing miR-134 on protein expression of MMP1.Results The results of dual luciferase reporter gene showed that miR-134 could bind to the 3′-UTR of MMP1 and significantly inhibited the luciferase activity.The cellular activity after 24 h in the miR-134 mimics group was significantly lower than that in the blank control group(P<0.01),while the miR-134 antagomir group was significantly higher than the blank control group(P<0.05).Compared with the apoptosis rate[(4.32±0.36)%]in the blank control group,(12.02±0.45)%in the miR-134 mimics group was significantly increased(P<0.01),while(2.31±0.26)%in the miR-134 antagomir group was significantly decreased(P<0.05).Compared with the blank control group,the cellular invasion and migration abilities in the miR-134 mimics group were significantly weakened and the MMP1 protein expression was significantly decreased(P<0.05),while which in the miR-134 antagomir group were significantly strengthened,and the MMP1 protein expression was significantly increased(P<0.05).Conclusion miR-134 affects the proliferation,apoptosis,invasion and migration of laryngeal cancer cells by regulating the expression of MMP1 protein at the post-transcriptional level.