TRIP13高表达对A549细胞增殖和侵袭的影响及其机制

High expression of TRIP13 promotes the proliferation and invasion of A549 cells and its mechanism

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.