miR-138对肾癌细胞增殖、迁移和侵袭的影响及其机制

Effects of miR-138 on proliferation,migration,and invasion of renal cancer cells

目的观察miR-138对肾癌细胞增殖、迁移和侵袭的影响,并探讨其对细胞周期蛋白D1(CCND1)的靶向调控作用。方法取肾癌细胞786-O,随机分为对照组、miR-138过表达组、miR-138低表达组,分别将miR-138空脂质体、miR-138 mimic和miR-138 inhibitor转染至细胞中。采用CCK-8法观察各组细胞增殖情况,细胞划痕实验观察细胞迁移能力,Transwell实验观察细胞侵袭能力。利用Star Base软件对miR-138进行靶基因预测,荧光素酶实验验证miR-138与CCND1的靶向调控关系。结果与对照组比较,miR-138过表达组细胞增殖活性降低,细胞迁移距离和细胞侵袭数量减少,CCND1蛋白表达下降(P均<0.01)。与对照组比较,miR-138低表达组细胞增殖活性升高,细胞迁移距离和细胞侵袭数量增加,CCND1蛋白表达升高(P均<0.01)。Star Base软件预测结果显示,CCND1的3'非编码区含有miR-138的结合位点。荧光素酶实验表明,miR-138可靶向调控CCND1。结论miR-138能够抑制肾癌细胞的增殖、迁移、侵袭,其机制可能与靶向调控CCND1表达有关。

Objective To observe the effects of miR-138 on proliferation,migration,and invasion of renal cancer cells,and to explore its targeted regulatory effect on cyclin D1(CCND1).Methods Kidney cancer cells 786-O were taken and randomly divided into the control group,miR-138 overexpression group,and miR-138 low expression group,which were transfected with miR-138 null liposome,miR-138 mimic,and miR-138 inhibitor,respectively.CCK-8 assay was used to observe the proliferation of the cells in each group,and the Scratch assay was used to observe the cell migration ability.Transwell assay was used to observe cell invasion ability.The target genes of miR-138 were predicted by StarBase software.The luciferase assay was utilized to determine the targeted regulatory relationship between miR-138 and CCND1.Results Compared with the control group,miR-138 overexpression group showed decreased cell proliferation activity,decreased distance of cell migration and the number of cell invasion,and decreased CCND1 protein expression(all P<0.01).Compared with the control group,the miR-138 low expression group showed elevated cell proliferative activity,increased distance of cell migration and the number of cell invasion,and elevated CCND1 protein expression(all P<0.01).StarBase software predictive result indicated that the 3'non-coding region of CCND1 contained the binding site of miR-138.Luciferase assay showed that miR-138 could regulate CCND1.Conclusion The miR-138 could inhibit the proliferation,migration and invasion of renal cancer cells,and its mechanism might be related to the targeted regulation of CCND1.