miR-122靶向调控CREB1对膀胱癌细胞增殖、侵袭能力的影响及其分子机制  

Effects and mechanism of miR-122 targeting CREB1 on proliferation and invasion of bladder cancer cells

作　　者：王艺璇[1] 郭立华[1] WANG Yixuan;GUO Lihua(China-Japan Union Hospital of Jilin University,Changchun 130033,China)

机构地区：[1]吉林大学中日联谊医院,长春130033

出　　处：《山东医药》2020年第35期26-29,共4页Shandong Medical Journal

基　　金：吉林省教育厅“十三五”科学技术项目(JJKH20190085KJ)。

摘　　要：目的观察微小RNA122(miR-122)通过靶向调控环磷腺苷效应元件结合蛋白1(CREB1)对膀胱癌细胞增殖和侵袭能力的影响,并探讨其可能的作用通路。方法TargetScan7.1软件预测程序显示,miR-122与CREB1启动子区存在连续的结合位点,双荧光素酶基因报告实验结果显示,在膀胱癌J82细胞中miR-122可靶向负调控CREB1。将J82细胞分为三组,共转染组采用Lipofectamine TM 2000转染miR-122 mimic及CREB1过表达质粒,miR-122 mimic组转染miR-122 mimic,对照组转染对照质粒,转染48 h。采用MTS法和平板克隆实验检测细胞增殖能力(分别以增殖OD值和克隆形成数表示),Transwell小室实验检测细胞侵袭能力(以穿膜细胞数表示),Western blotting法检测细胞CREB1及蛋白激酶B(Akt)、磷酸化Akt(p-Akt)、雷帕霉素靶蛋白(mTOR)、磷酸化mTOR(p-mTOR)蛋白表达。结果与对照组比较,miR-122 mimic组和共转染组细胞增殖OD值、克隆形成数、穿膜细胞数均降低,且miR-122 mimic组降低更明显(P均<0.05)。与对照组比较,miR-122 mimic组和共转染组细胞CREB1、p-Akt、p-mTOR蛋白表达降低,且miR-122 mimic组降低更明显(P均<0.05)。与对照组比较,miR-122 mimic组和共转染组细胞Akt、mTOR表达均无明显变化(P均>0.05)。结论miR-122通过作用靶点CREB1负向调控Akt/mTOR通路,从而抑制膀胱癌细胞的增殖和侵袭。Objective To observe the effects of microRNA(miR-122)on the proliferation and invasion of bladder cancer cells by targeting cyclic adenosine phosphate response element binding protein-1(CREB1),and to explore the possible pathway of action.Methods The prediction program of targetscan 7.1 software showed that there was a binding site between miR-122 and CREB1 promoter.The results of double luciferase gene reporting showed that miR-122 could target and negatively regulate CREB1 in the bladder cancer J82 cells.J82 cells were divided into three groups.The cells in the co-transfection group were transfected with miR-122 mimic and CREB1 overexpression plasmids with Lipofectamine TM2000,the miR-122 mimic group with miR-122 mimic,and the control group with the control plasmid for 48 h.MTS and plate cloning experiments(represented by OD value and clone formation number,respectively)were used to detect the cell proliferation ability.Transwell assay(represented by the number of transmembrane cells)was used to detect the cell invasion ability.Western blotting was used to detect the expression of CREB1 and protein kinase B(Akt),phosphorylated Akt(p-Akt),rapamycin target protein(mTOR),phosphorylated mTOR(p-mTOR)protein.Results Compared with the control group,the OD value,clone formation number and the number of transmembrane cells in the miR-122 mimic group and co-transfection group decreased,and the decrease was more significant in the miR-122 mimic group(all P<0.05).Compared with the control group,the expression of CREB1,p-Akt and p-mTOR in the miR-122 mimic group and co-transfection group decreased,with more significant decrease in the miR-122 mimic group(all P<0.05).Compared with the control group,the expression of Akt and mTOR in the miR-122 mimic group and co-transfection group had no significant change.Conclusion The miR-122 negatively regulates Akt/mTOR pathway by targeting CREB1,thereby inhibiting proliferation and invasion of bladder cancer cells.

关 键 词：膀胱癌 MIR-122 环磷腺苷效应元件结合蛋白1 细胞增殖 细胞侵袭 

分 类 号：R737.14[医药卫生—肿瘤]