摘要
目的探讨ITCH在肺癌细胞系中的表达以及ITCH沉默对肺癌A549细胞增殖、侵袭、凋亡的影响及其可能机制。方法采用Western blotting检测人正常肺上皮细胞系BEAS-2B和人肺癌细胞系A549、Calu3中ITCH蛋白的表达情况。采用PEI转染试剂分别向A549细胞转染si ITCH(si ITCH组)和si NC(si NC组),采用CCK-8法、Transwell实验和流式细胞术检测两组细胞增殖能力、侵袭能力和凋亡的变化,Western blotting检测两组ITCH、Bcl-2、Bax、Cyclin D1、MMP-2、MMP-9、β-catenin、E-cadherin蛋白表达。结果人肺正常上皮细胞系BEAS-2B中ITCH蛋白表达水平为0. 98±0. 043,显著低于Calu3细胞的1. 92±0. 073和A549细胞的2. 74±0. 151,差异均有统计学意义(P <0. 05)。si ITCH组96 h时细胞增殖倍数为1. 97±0. 021,显著低于si NC组的3. 72±0. 232(P <0. 05); si ITCH组细胞凋亡率为(43. 2±1. 52)%,显著高于si NC组的(2. 3±0. 32)%,差异有统计学意义(P <0. 05)。si ITCH组穿膜细胞数为(297. 2±22. 21)个,显著低于si NC组的(601. 2±3. 21)个,差异有统计学意义(P <0. 05)。与si NC组比较,si ITCH组Bcl-2、Cyclin D1、MMP-2、MMP-9、β-catenin表达水平降低,Bax、E-cadherin表达水平升高。结论 ITCH沉默通过调控MMP、Cyclin D1、Bcl-2/Bax等信号通路抑制肺癌细胞增殖、侵袭并诱导其凋亡。
Objective To investigate the expression of ITCH in lung cancer cells and the effect of ITCH silencing on proliferation,invasion and apoptosis of lung cancer cells A549 and its possible mechanisms.Methods Western blotting was used to detect the expression of ITCH protein in human normal lung epithelial cell lines BEAS-2B and human lung cancer cell lines A549 and Calu3.SiITCH(siITCH group)and siNC(siNC group)were transfected into A549 cells by PEI transfection reagent,respectively.CCK-8 method,Transwell test and flow cytometry were used to detect the changes of cell proliferation,invasion and apoptosis in the two groups.The protein expression of ITCH,Bcl-2,Bax,Cyclin D1,MMP-2,MMP-9,β-catenin and E-cadherin in the two groups were detected by Western blotting assay.Results The expression level of ITCH protein in human lung normal epithelial cell line BEAS-2B was 0.98±0.043,significantly lower than that in Calu3 cell line(1.92±0.073)and A549 cell line(2.74±0.151),and the difference was statistically significant(P<0.05).The cell proliferation rate in siITCH group was 1.97±0.021 at 96 h,which was significantly lower than that in siNC group(3.72±0.232),and the difference was statistically significant(P<0.05).The apoptotic rate in siITCH group was(43.2±1.52)%,which was significantly higher than that in siNC group(2.3±0.32)%.The difference was statistically significant(P<0.05).The number of perforating cells in siITCH group was 297.2±21.21,which was significantly lower than 601.2±3.21 in siNC group(P<0.05).Compared with the siNC group,the expression levels of Bcl-2,Cyclin D1,MMP-2,MMP-9 andβ-catenin in siITCH group decreased,while the expression levels of Bax and E-cadherin increased.Conclusion Silencing ITCH inhibits lung cancer proliferation,invasion and induces apoptosis by regulating signaling pathways such as MMP,Cyclin D1 and Bcl-2/Bax.
作者
蓝冰
张东伟
孔晋亮
钟家将
贺婵娟
LAN Bing;ZHANG Dongwei;KONG Jinliang;ZHONG Jiajiang;HE Chanjuan(Department of Respiratory and Critical Care Medicine,Liuzhou People's Hospital,Liuzhou 545000,China)
出处
《临床肿瘤学杂志》
CAS
北大核心
2019年第1期16-20,共5页
Chinese Clinical Oncology
基金
广西科技计划资助项目(桂科AB16380152)
关键词
肺癌
ITCH
增殖
凋亡
侵袭
Lung cancer
ITCH
Proliferation
Apoptosis
Invasion
