Mindbomb同源物2基因转染对人脑胶质瘤细胞U251增殖的影响及其机制  

Effects of Mindbomb homologue 2 gene transfection on proliferation of human glioma U251 cells

作　　者：单小松[1] 刘海鹏[1] 郑克彬[1] 方川[1] 丁亚楠[1] 

机构地区：[1]河北大学附属医院,河北保定071000

出　　处：《山东医药》2016年第43期4-6,共3页Shandong Medical Journal

基　　金：河北省临床医学优秀人才培养和基础课题研究项目(361007)

摘　　要：目的观察Mindbomb同源物2(MIB2)基因转染对人脑胶质瘤细胞U251增殖的影响,并探讨其作用机制。方法将U251细胞分为空白对照组、阴性对照组、实验组1、实验组2;空白对照组不作任何处理,阴性对照组转染空载体FLAG质粒,实验组1转染MIB2-FLAG质粒,实验组2转染si MIB2。用PCR法检测各组细胞的MIB2mRNA,用蛋白质印迹法检测各组细胞的MIB2、核因子κB(NF-κB)抑制性蛋白(IκB)、NF-κB蛋白,用MTT法检测各组细胞的光密度值。结果实验组1的MIB2 mRNA相对表达量为19.43±3.21、实验组2为0.43±0.17、阴性对照组为1.03±0.12;实验组1、实验组2与阴性对照组相比,P均<0.01。实验组1的MIB2、核因子κB(NF-κB)抑制性蛋白、NF-κB蛋白相对表达量分别为5.11±0.41、0.49±0.13、4.99±0.32,实验组2分别为0.43±0.17、3.12±0.36、0.49±0.14,阴性对照组分别为1.09±0.11、1.02±0.46、1.01±0.21;实验组1、实验组2与阴性对照组相比,P均<0.01;在36、48、72 h三个时点,与阴性对照组和空白对照组相比,实验组1的U251细胞光密度值升高(P均<0.01)。结论转染MIB2基因后人脑胶质瘤U251细胞的增殖能力增强,MIB2基因可能是通过MIB2蛋白调控NF-κB信号通路而发挥作用。Objective To observe the effects of Mindbomb homologue 2 （MIB2）gene transfection on the proliferation of human glioma U251 cells,and to explore its mechanism.Methods U251 cells were divided into four groups：the con-trol group,negative control group,experimental group 1 and experimental group 2.U251 cells in the control group was un-treated;U251 cells in the negative control group was transfected with vector FLAG plasmid;and U251 cells in the experi-mental group 1 was transfected with MIB2-FLAG plasmid;U251 cells in the experimental group 2 was transfected siMIB2. MIB2 mRNA was detected by PCR,the expression of MIB2,nuclear factor-κB （NF-κB）inhibitory protein （IκB）and NF-κB protein was detected by Western blotting in each group,and the optical density （OD）values were detected by MTT.Results The expression of MIB2 mRNA in the experimental group 1,experimental group 2 and negative control group was 19.43 ±3.21,0.43 ±0.17,and 1.03 ±0.12,respectively;and significant difference was found in the MIB2 mRNA between the experimental groups 1,2 and the negative control group （all P〈0.01）.The expression of MIB2,IκB and NF-κB protein in the experimental group 1 was 5.11 ±0.41,0.49 ±0.13 and 4.99 ±0.32,respectively;the expression of MIB2,IκB and NF-κB protein in the experimental group 2 was 0.43 ±0.17,3.12 ±0.36 and 0.49 ±0.14,respectively;and the expression of MIB2,IκB and NF-κB protein in the negative control group was 1.09 ±0.11,1.02 ±0.46 and 1.01 ± 0.21,respectively.Significant difference was found in the expression of MIB2,IκB and NF-κB protein between the experi-mental groups 1,2 and the negative control group （all P〈0.01）.At 36,48 and 72 h,compared with the negative control group and blank control group,OD value of U251 cells in the experimental group 1 increased significantly （all P〈0.01）. Conclusion After MIB2 gene tranfection,the proliferation of human glioma U251 cells increases and MIB 2 gene may play an important role in the regulation of NF-κB signalin

关 键 词：Mindbomb同源物2 核因子KB 核因子KB抑制性蛋白 脑胶质瘤 细胞增殖 

分 类 号：R739.41[医药卫生—肿瘤]