摘要
目的探讨甲状腺激素受体相互作用蛋白(TRIP13)在非小细胞肺癌细胞增殖和侵袭中的作用和机制。方法蛋白质印迹法检测2015-01-01-2018-12-31在中国医科大学附属第一医院行肺癌手术的24例非小细胞肺癌组织和癌旁正常肺组织中TRIP13的表达。根据基因转染或siRNA干扰改变肺癌A549细胞中TRIP13的表达量。实验划分为4组:转染对照组(NC)、转染组(TRIP13)、干扰对照组(siNC)和干扰组(siTRIP13),并用蛋白质印迹检测E-cadherin、N-cadherin、Vimentin和Snail的表达。CCK-8、克隆形成和侵袭实验检测细胞增殖和侵袭能力。结果TRIP13在正常肺组织和肺癌组织中的表达量分别是2.06±0.70和3.24±1.31,差异有统计学意义,n=24,t=3.83,P<0.001。转染TRIP13后,与NC组比较,A549细胞中TRIP13、N-cadherin、Vimentin、Snail和E-cadherin的相对表达量分别是1.90±0.12、2.12±0.19、1.38±0.08、1.70±0.12和0_65±0.08,均值差异均有统计学意义,t_(TRIP13)=15.12,P=0.01;t_(N-cadherin)=9.86,P=0.01;t_(Vimentin)=7.77,P=0.02,t_(Snail)=7.11,P=0.02,t_(E-cadherin)=9.64,P=0.01.癌细胞的增殖能力升高,与NC组比较差异有统计学意义,并呈时间依赖,F_(分组)=99.30,F_(时间)=341.31,F_(分组×时间=)10.90,均P<0.001;克隆形成数量(TRIP13组为76.00±8.98,NC组为43.33±6.34;t=4.83,P=0.04,n=3)和侵袭能力(TRIP13组为147.67±7.04,NC组为29.67±7.41;t=14.17,P=0.01,n=3)均高于NC组,差异均有统计学意义。干扰TRIP13后,TRIP13、N-cad-herin、Vimentin、Snail和E-cadherin的表达量分别是0.66±0.08、0.70±0.08、0.65±0.07、0.62±0.06和2.04±0.12,差异均有统计学意义,t_(TRIP13)=6.20,P=0.03;t_(N-cadherin)=5.28,P=0.03;t_(Vimentin)=6.70,P=0,02;t_(Snail)=7.85,P=0.02;t_(E-cadherin)=8.39,P=0.01。A549细胞的增殖能力下降,F_(分组)=70.00,F_(时间)=426.05,F_(分组×时间)=7.76,均P<0.001。同时,siTRIP13组A549细胞的克隆形成数量为72.33±6.13,NC组为113.00±10.42,t=5.46,P=0.03,n=3。siTRIP13组侵袭能力为35.67±3.30,NC组为190.33±12.76,t=20.53,P=0.01,n=3。结论TRIP13在非小细胞肺癌组织中高表达,并可促进上皮间质转化(EMT)和肺癌细胞的增殖和侵袭。
Objective To investigate the role and mechanism of thyroid hormone receptor interactor 13(TRIP13)in the proliferation and invasion of non-small cell lung cancer(NSCLC).METHODS Western blotting was used to detect the expression of TRIP13 in 24 pairs of non-small cell lung cancer tissues and normal lung tissues undergoing lung cancer surgery in the First Affiliated Hospital of China Medical University from 2015-01-01 to 2018-12-31;According to gene transfection or siRNA interference to change the expression of TRIP13 in lung cancer A549 cells.The experiment was divided into four groups:transfection control group(NC),transfection group(TRIP13),interference control group(siNC)and interference group(siTRIP13),and Western blot was used to detect the expression of E-cadherin,N-cadherin,Vimentin and Snail.CCK-8,colony formation assay and invasion assay were used to detect cell proliferation and invasion.RESULTS The expression levels of TRIP13 in normal lung tissue and lung cancer tissue were 2.06±0.70 and 3.24±1.31,the difference was statistically significant(n=24,t=3.83,P<0.001).Compared with the transfection control group,after TRIP13 was transfected,the relative expression levels of TRIP13,N-cadherin,Vimentin,Snail and E-cadherin in A549 cells were 1.90±0.12,2.12±0.19,1.38±0.08,1.70±0.12 and 0.65±0.08.the mean difference is statistically significant(tTRIP13=15.12,P=0.01;tN-cadherin=9.86,P=0.01;tVimentin=7.77,P=0.02;tSnail=7.11,P=0.02;tE-cadherin=9.64,P=0.01).The proliferation ability of cancer cells increased,and the difference was statistically significant compared with the transfection control group,and it was time-dependent.Fgroup=99.30,Ftime=341.31,Fgroup×time=10.90,both P<0.001;The number of clones formed(transfection group:76.00±8.98,transfection control group:43.33±6.34;t=4.83,P=0.04;n=3)and invasion ability(transfection group:147.67±7.04,transfection control group:29.67±7.41;t=14.17,P=0.01;n=3)are higher than those of the transfection control group,the differences are statistically significant.After interference with TRIP13,the expression levels of TRIP13,N-cadherin,Vimentin,Snail,and E-cadherin were 0.66±0.08,0.70±0.08,0.65±0.07,0.62±0.06,and 2.04±0.12,respectively.The difference was statistically significant(tTRIP13=6.20,P=0.03;tN-cadherin=5.28,P=0.03;tVimentin=6.70,P=0.02;tSnail=7.85,P=0.02;tE-cadherin=8.39,P=0.01).The proliferation ability of A549 cells decreased(Fgroup=70.00,P<0.001;Ftime=426.05,P<0.001;Fgroup×time=7.76,both P<0.001).At the same time,the number of clones formed of A549 cells in the siTRIP13 group was 72.33±6.13,and in the NC group was 113.00±10.42(t=5.46;P=0.03;n=3).The invasive ability of siTRIP13 group was 35.67±3.30,and in NC group was 190.33±12.76(t=20.53;P=0.01;n=3),the difference is statistically significant.CONCLUSION TRIP13 is highly expressed in NSCLC tissues,and can promote EMT process and the proliferation and invasion of NSCLC cells.
作者
刘晨晨
李志瀚
雷蕾
黄文镜
徐洪涛
LIU Chen-chen;LI Zhi-han;LEI Lei;HUANG Wen-jing;XU Hong-tao(Department of Pathology,First Hospital of China Medical University,Shenyang 110001,China;Department of Pathology,College of Basic Medical Sciences,China Medical University,Shenyang 110122,China)
出处
《中华肿瘤防治杂志》
CAS
北大核心
2021年第13期1001-1006,共6页
Chinese Journal of Cancer Prevention and Treatment
基金
辽宁省自然科学基金(2020-MS-179)。
关键词
肺癌
TRIP13
上皮间质转化
增殖
侵袭
lung cancer
TRIP13
epithelial-mesenchymal transition
proliferation
invasion