通用转录因子3表达对食管鳞癌细胞增殖及凋亡的影响  

Influence of basic transcription factor 3 on proliferation and apoptosis of esophageal squamous cell carcinoma cells

作　　者：刘清 刘涛[2] 郑树涛 阿尔孜古丽·吐尔逊 谭依依 梁艳 卢晓梅 LIU Qing;LIU Tao;ZHENG Shutao;Aerziguli TUERXUN;TAN Yiyi;LIANG Yan;LU Xiaomei(State Key Laboratory of Pathogenesis,Prevention,Treatment of High Incidence Diseases in Central Asia,Clinical Medical Research Institute,the First Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang UygurAutonomous Region 8300ll,China;Department of Clinical Laboratory,the First Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang Uygur Autonomous Region 830054,China)

机构地区：[1]新疆医科大学第一附属医院临床医学研究院、省部共建中亚高发病成因与防治国家重点实验室,新疆维吾尔自治区乌鲁木齐830011 [2]新疆医科大学第一附属医院检验科,新疆维吾尔自治区乌鲁木齐830054

出　　处：《中华实用诊断与治疗杂志》2023年第8期776-780,共5页Journal of Chinese Practical Diagnosis and Therapy

基　　金：新疆维吾尔自治区自然科学基金杰出青年科学基金(2021D01E31);新疆医科大学、省部共建国家重点实验室国家优秀青年科学基金培育项目(xyd2021y006);省部共建中亚高发病成因与防治国家重点实验室开放课题(SKL-HIDCA-2020-SG2);国家自然科学基金(81860511)。

摘　　要：目的观察通用转录因子3(BTF3)在食管鳞癌细胞中表达情况,探讨BTF3对食管鳞癌细胞增殖、凋亡的影响及可能机制。方法取对数生长期正常食管鳞状上皮细胞株Het-1A、食管鳞癌细胞株KYSE150,采用Western blot法检测BTF3蛋白相对表达量。对数生长期KYSE150细胞分为BTF3敲低组(转染BTF3敲低慢病毒)、TWIST1过表达组(转染BTF3敲低+TWIST1过表达慢病毒)、基质金属蛋白酶-2(MMP-2)过表达组(转染BTF3敲低+MMP-2过表达慢病毒)、对照组(转染阴性对照慢病毒)。BTF3敲低组、对照组细胞采用实时荧光定量PCR法检测BTF3 mRNA相对表达量,采用Western blot法检测BTF3、MMP-2、TWIST1、β-catenin、p-β-catenin蛋白相对表达量,采用流式细胞术检测细胞凋亡率。转染48、72、96、120 h时,应用Celigo细胞成像分析仪检测4组细胞增殖倍数。结果KYSE150细胞BTF3蛋白相对表达量(1.42±0.14)高于Het-1A细胞(1.09±0.10)(t=3.289,P=0.030)。BTF3敲低组细胞BTF3 mRNA和蛋白相对表达量(0.04±0.01、0.75±0.12)均低于对照组(1.25±0.44、1.03±0.05)(t=4.756,P=0.009;t=3.645,P=0.022),细胞凋亡率[(13.01±0.10)%]高于对照组[(3.34±0.14)%](t=97.670,P<0.001);BTF3敲低组细胞MMP-2、TWIST1、p-β-catenin蛋白相对表达量(0.65±0.02、0.92±0.04、0.56±0.08)均低于对照组(1.05±0.05、1.09±0.08、0.99±0.02)(t=13.350,P<0.001;t=3.162,P=0.034;t=9.360,P<0.001),β-catenin蛋白相对表达量(0.91±0.10)与对照组(0.97±0.07)比较差异无统计学意义(t=1.152,P=0.314)。转染48、72、96、120 h时,BTF3敲低组、TWIST1过表达组、MMP-2过表达组细胞增殖倍数均低于对照组(P<0.05),TWIST过表达组均高于BTF3敲低组(P<0.05),BTF3敲低组与MMP-2过表达组比较差异均无统计学意义(P>0.05)。结论敲低食管鳞癌细胞中BTF3表达可抑制β-catenin磷酸化及TWIST1表达,从而抑制上皮-间质转化和细胞增殖,促进细胞凋亡。Objective To observe the expression of basic transcription factor 3(BTF3)in esophageal squamous cell carcinoma(ESCC),and to investigate the influence of BTF3 on the proliferation and apoptosis of ESCC cells and its mechanism.Methods The normal esophageal squamous epithelial cell line Het-1A and ESCC cell line KYSE150 in logarithmic growth phase were collected to detect the relative expression of BTF3 protein by Western blot.The KYSE150cells in logarithmic growth phase were divided into BTF3 knockdown group(transfected with BTF3 knockdown lentivirus),TWIST1 overexpression group(transfected with BTF3 knockdown and TWIST1 overexpression lentivirus),matrix metalloproteinase-2(MMP-2)overexpression group(transfected with BTF3 knockdown and MMP-2overexpression lentivirus),and control group(transfected with negative control lentivirus).In BTF3 knockdown group and control group,the relative expression of BTF3 mRNA was detected by real-time fluorescence quantitative PCR,the relative expressions of BTF3,MMP-2,TWIST1,β-catenin and p-β-catenin proteins were detected by Western blot,and the apoptotic rate was detected by flow cytometry.At 48,72,96 and 120 h of transfection,Celigo cell imaging analyzer was used to detect the proliferation fold in four groups.Results The relative expression of BTF3 protein was significantly higher in KYSE150 cells(1.42±0.14)than that in Het-1A cells(1.09±0.10)(t=3.289,P=0.030).The relative expressions of BTF3 mRNA and protein were lower in BTF3 knockdown group(0.04±0.01,0.75±0.12)than those in control group(1.25±0.44,1.03±0.05)(t=4.756,P=0.009;t=3.645,P=0.022),and the apoptotic rate was higher in BTF3 knockdown group[(13.01±0.10)%]than that in control group[(3.34±0.14)%](t=97.670,P<0.001).The relative expressions of MMP-2,TWIST1 and p-β-catenin proteins were lower in BTF3 knockdown group(0.65±0.02,0.92±0.04,0.56±0.08)than those in control group(1.05±0.05,1.09±0.08,0.99±0.02)(t=13.350,P<0.001;t=3.162,P=0.034;t=9.360,P<0.001),and there was no significant difference inβ-catenin pr

关 键 词：食管鳞癌 通用转录因子3 增殖 凋亡 TWIST1 Β-CATENIN 

分 类 号：R735.1[医药卫生—肿瘤]