let-7b-5p靶向IGF1R调控脑胶质瘤细胞生长的机制  

Mechanism of let-7b-5p targeting IGF1R to inhibit glioma cell growth

作　　者：刘喜红[1] 杜晓丹 范孟杨 徐柳清 杨丽萍[1] 侯俊林[1] 赵培源[1] Liu Xihong;Du Xiaodan;Fan Mengyang;Xu Liuqing;Yang Liping;Hou Junlin;Zhao Peiyuan(Henan University of Chinese Medicine,Traditional Chinese Medicine/Zhongjing School,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学中医学院(仲景学院),郑州450046

出　　处：《临床与实验病理学杂志》2025年第3期359-364,共6页Chinese Journal of Clinical and Experimental Pathology

基　　金：国家自然科学基金(82004108);河南省科技攻关项目(242102310385);河南省高等学校青年骨干教师培养计划(2023GGJS082);河南省博士后科研资助项目(HN2024094)。

摘　　要：目的探讨let-7b-5p在脑胶质瘤中的表达及其对U251细胞生长的影响和潜在的作用机制。方法采用qRT-PCR法检测let-7b-5p在脑胶质瘤细胞和组织中的表达量,通过CGGA数据库分析let-7b-5p表达与脑胶质瘤WHO分级和总生存率的关系;通过瞬时转染法下调脑胶质瘤细胞U251中let-7b-5p和胰岛素样生长因子1受体(insulin-like growth factor 1 receptor,IGF1R)的表达,采用qRT-PCR、CCK8、克隆形成实验、Western blot、双荧光素酶报告基因等技术检测let-7b-5p在U251细胞生长中的作用及潜在的机制。结果let-7b-5p在脑胶质瘤细胞(A172:3.64±0.64、V251:4.56±0.52、U87-MG:3.31±0.50)和组织中(2.18±0.22)的表达显著高于星形胶质细胞(HMC3:1.00±0.21,P<0.05或P<0.01)和正常脑组织(1.01±0.19,P<0.05),其表达水平与脑胶质瘤患者WHO分级呈负相关(P<0.0001),与原发性和复发性脑胶质瘤患者的生存率呈正相关(P<0.0001,P=0.028);敲减U251细胞中let-7b-5p的表达可促进脑胶质瘤细胞的生长(CCK8:let-7b-5p敲减组126.00±12.09 vs miR-NC组90.93±5.13,P<0.05),激活PI3K/AKT信号通路;在U251细胞中抑制IGF1R表达可以逆转let-7b-5p低表达促进脑胶质瘤细胞生长[CCK8:let-7b-5p敲减组+IGF1R敲减组(92.08±6.14)vs let-7b-5p敲减组+sh-NC组(116.67.08±8.50)]、激活PI3K/AKT信号通路的效应。结论let-7b-5p在脑胶质瘤细胞中起抑癌基因作用,let-7b-5p可能通过靶向IGF1R激活PI3K/AKT信号通路参与脑胶质瘤细胞的进程。Purpose To explore the expression level of let-7b-5p in glioma and its effects and potential mechanisms on U251 cell growth.Methods The expression of let-7b-5p in glioma was detected using qRT-PCR.Data from the CGGA database were analyzed to examine the relationship between the let-7b-5p expression levels,WHO grade and overall survival rates of glioma patients.Transient transfection was used to downregulate the expression of let-7b-5p and IGF1R in U251 cells.The role and potential mechanism of let-7b-5p in the U251 cell were evaluated using qRT-PCR,CCK8 assays,clone formation assays,Western blotting,and double luciferase reporter assays.Results The expression of let-7b-5p in glioma cells(A172:3.64±0.64,V251:4.56±0.52,U87-MG:3.31±0.50)and tissues(2.18±0.22)was significantly higher than that in astrocytes(HMC3:1.00±0.21,P<0.05 or P<0.01)and normal brain tissues(1.01±0.19,P<0.05).Let-7b-5p expression was negatively correlated with WHO grades but positively correlated with survival rates in primary and recurrent glioma patients(P<0.0001 and P=0.028,respectively).Knockdown of let-7b-5p in U251 cells significantly promoted the growth of glioma cells(CCK8:knockdown group 126.00±12.09 vs miR-NC group 90.93±5.13,P<0.05)and activated PI3K/AKT signal pathway.Suppressing IGF1R expression in U251 cells reversed the effects of let-7b-5p knockdown on glioma cell growth[CCK8:let-7b-5p knockdown+IGF1R knockdown group(92.08±6.14)vs let-7b-5p knockdown+sh-NC group(116.67.08±8.50)]and PI3K/AKT signal pathway activation.Conclusion Let-7b-5p functions as a tumor suppressor gene in glioma.It may regulate glioma cell growth by targeting IGF1R and modulating PI3K/AKT signal pathway.

关 键 词：脑胶质瘤 let-7b-5p IGF1R PI3K/AKT 

分 类 号：R739.41[医药卫生—肿瘤]