沉默Gab1抑制人胆管癌TFK-1细胞增殖的可能机制  

Mechanism of silencing Gab1in suppressing the proliferation of human cholangiocarcinoma TFK-1cells

作　　者：王刚[1] 桑海泉[2] 毕聪[3] WANG Gang;SANG Haiquan;BI Cong(Department of Radiology,the Fourth Affiliated Hospital of China Medical University,Shenyang110032,China;Department of General Surgery,the Fourth Affiliated Hospital of China Medical University,Shenyang110032,China;Department of Radiology,the First Affiliated Hospital of China Medical University,Shenyang110001,China)

机构地区：[1]中国医科大学附属第四医院放射科,沈阳110032 [2]中国医科大学附属第四医院普通外科,沈阳110032 [3]中国医科大学附属第一医院放射科,沈阳110001

出　　处：《中华实用诊断与治疗杂志》2020年第5期441-444,共4页Journal of Chinese Practical Diagnosis and Therapy

基　　金：辽宁省科学技术计划项目(201501018)。

摘　　要：目的探讨沉默Grb2相关结合蛋白1(Grb2-associated binding proteins 1,Gab1)提高放射线照射效果对人胆管癌TFK-1细胞增殖的抑制作用及可能机制。方法对数生长期人胆管癌TFK-1细胞随机分为放射组(转染小干扰RNA-NC质粒)、观察组(转染小干扰RNA-Gab1质粒)和对照组(转染小干扰RNA-NC质粒)。转染后培养48h,放射组、观察组应用8Gy X线照射24h,对照组不进行X线照射。取X线照射24h的细胞,采用CCK-8法检测细胞活力,平板细胞克隆形成实验检测细胞增殖能力,实时荧光定量PCR法检测Gab1mRNA相对表达量,Western Blot法检测Gab1、PI3K、Akt蛋白相对表达量。结果X线照射24h,放射组、观察组细胞活力吸光度值(0.52±0.05、0.41±0.04)、细胞增殖形成的菌落数[(214.32±6.47)、(164.34±6.03)个]、Gab1mRNA相对表达量(4.05±0.36、0.83±0.18)、Gab1蛋白相对表达量(3.05±0.36、2.03±0.18)、PI3K蛋白相对表达量(1.31±0.10、0.83±0.18)、Akt蛋白相对表达量(3.54±0.22、1.15±0.18)均低于对照组[0.65±0.04、(289.54±8.53)个、5.55±0.21、3.85±0.21、1.96±0.09、4.36±0.24](P<0.05),且观察组低于放射组(P<0.05)。结论沉默Gab1可提高放射线照射对人胆管癌TFK-1细胞活力及增殖的抑制作用,其机制可能与下调PI3K/Akt表达有关。Objective To investigate the role of silencing Grb2-associated binding protein 1(Gab1)in inhibiting the proliferation of cholangiocarcinoma cells by increasing radiation exposure and its possible mechanism.Methods Human cholangiocarcinoma TFK-1cells in logarithmic growth phase were randomly divided into radiation group,observation group and control group,transfected with small interfering RNA empty plasmid,small interfering RNA Gab1plasmid and small interfering RNA empty plasmid,respectively.After 48htransfection,radiation group and observation group received 8Gy X-ray radiation for 24h,and control group received no radiation.After 24hradiation,the cell viability was measured by CCK-8method,the cell proliferation was detected by clone forming assay,the relative expression of Gab1 mRNA was detected by real-time quantitative PCR,and the relative expressions of Gab1,PI3Kand Akt were detected by Western blot.Results The optical density value(0.65±0.04,0.52±0.05,0.41±0.04),the colony forming unit(289.54±8.53,214.32±6.47,164.34±6.03),the relative expression of Gab1mRNA(5.55±0.21,4.05±0.36,0.83±0.18),the relative expression of Gab1protein(3.85±0.21,3.05±0.36,2.03±0.18),the relative expression of PI3Kprotein(1.96±0.09,1.31±0.10,0.83±0.18)and the relative expression of Akt protein(4.36±0.24,3.54±0.22,1.15±0.18)decreased gradually in turn in control group,radiation group and observation group(P<0.05).Conclusion Silencing Gab1can increase the role of radiation exposure in inhibiting the activity and proliferation of TFK-1,probably by down-regulating PI3K/Akt expression.

关 键 词：胆管癌 放射 放疗 Grb2相关结合蛋白1 PI3K/AKT 

分 类 号：R735.8[医药卫生—肿瘤]