Smad7通过PI3K/Akt/mTOR信号通路调控人脑胶质瘤U-87MG细胞的生物学性状  

Smad7 regulates the biological characters of human glioma U-87MG cells through PI3K/Akt/mTOR signaling pathway

作　　者：谢云鹏[1] 胡军[1] 柳新 刘兵 于淼[1] 呼铁民[1] XIE Yunpeng;HU Jun;LIU Xin;LIU Bing;YU Miao;HU Tiemin(Department of Neurosurgery,Affiliated Hospital of Chengde Medical University,Chengde 067000,China;Department of Oncology,Affiliated Hospital of Chengde Medical University)

机构地区：[1]承德医学院附属医院神经外科,承德067000 [2]承德医学院附属医院肿瘤科

出　　处：《山西医科大学学报》2021年第6期689-694,共6页Journal of Shanxi Medical University

基　　金：河北省承德市科学技术研究与发展计划项目(自筹经费)(201804A035)。

摘　　要：目的探讨敲低Smad7对人脑胶质瘤U-87MG细胞增殖和凋亡的影响及机制。方法将U-87MG细胞分为:干扰组、阴性对照组和空白对照组,其中干扰组通过脂质体转染法转染靶向Smad7的siRNA,阴性对照组转染阴性对照siRNA,空白对照组不做任何处理。转染48 h采用Western blot检测Smad7的表达情况,转染后24,48,72 h进行细胞计数以绘制细胞增殖曲线,转染后48 h进行CCK-8实验以检测细胞增殖活力,流式细胞术检测各组细胞凋亡率,Western blot检测PCNA、CyclinD1、Bcl-2、Bax及PI3K/Akt/mTOR信号通路相关蛋白的表达情况。结果与阴性对照组相比,干扰组脑胶质瘤U-87MG细胞增殖能力均增强(P<0.05),细胞增殖相关蛋白PCNA和CyclinD1的表达均升高(P<0.05);细胞凋亡率降低(P<0.05),Bcl-2与Bax的比值降低(P<0.05);p-PI3K、p-Akt和p-mTOR的表达均升高(P<0.05)。结论敲低Smad7可通过PI3K/Akt/mTOR信号促进脑胶质瘤细胞的增殖并抑制其凋亡,Smad7有望成为脑胶质瘤生物治疗的备选靶点。Objective To investigate the effect of Smad7 knockdown on the proliferation and the apoptosis of human glioma U-87MG cells and its mechanism.Methods U-87MG cells were divided into three groups:interference group(transfected with siRNA target-ing Smad7),negative control group(transfected with negative control siRNA)and blank control group.The expression of Smad7 was detected 48 h after transfection by Western blot.Cell count was recorded to plot the cell proliferation curve at 24 h,48 h and 72 h after transfection.The cell proliferation activity was detected at 48 h after transfection by CCK-8 assay,the apoptosis rate of cells in each group was detected by flow cytometry,and the expression levels of PCNA,CyclinD1,Bcl-2,Bax and proteins related to PI3K/Akt/mTOR signaling pathway were detected by Western blot.Results Compared with negative control group,the proliferation capacity of glioma U-87MG cells was increased in interference group(P<0.05),the expression levels of proliferation-related proteins PCNA and CyclinD1 were both increased(P<0.05),the apoptosis rate and the the ratio of Bcl-2 to Bax were decreased(P<0.05),and the expression levels of p-PI3K,p-Akt and p-mTOR were increased(P<0.05).Conclusion Smad7 can inhibit the proliferation and promote the apoptosis of glioma cells through PI3K/Akt/mTOR signaling pathway,and it is expected to be an alternative target for the biotherapy of glioma.

关 键 词：SMAD7 脑胶质瘤 U-87MG 增殖 凋亡 PI3K/Akt/mTOR信号通路 

分 类 号：R739.4[医药卫生—肿瘤]