CAJ | 학술논문

目的探讨CD146对获得性Erlotinib耐药肺腺癌细胞上皮-间充质转化(EMT)的作用及机制.方法采用不同浓度的Erlotinib作用于肺腺癌细胞株H1650,诱导其产生耐药(H1650ER);免疫荧光法及Western blot检测耐药前后上皮型钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、CD146的表达变化;小干扰RNA(siRNA)干扰H1650ER细胞中CD146的表达,利用Transwell侵袭小室及细胞计数试剂盒(CCK-8)实验分别评价肺腺癌细胞侵袭及增殖能力,Western blot 检测相关蛋白的表达变化.结果获得性Erlotinib耐药肺腺癌细胞H1650ER中CD146及Vimentin 蛋白的表达量分别为0.474±0.013和0.465±0.012,显著高于耐药前(P＜0.05);而E-cadherin蛋白的表达量为0.180±0.011,明显低于耐药前(P＜0.05).CD146-siRNA干扰H1650ER细胞CD146的表达后可明显抑制其增殖能力,与对照组比较差异有统计学意义(P＜0.05);CD146-siRNA干扰后细胞侵袭力亦下降(P＜0.05).结论获得性Erlotinib耐药肺腺癌产生EMT并CD146表达显著上调;抑制CD146的表达可下降其侵袭及增殖能力.
목적 탐토CD146대획득성Erlotinib내약폐선암세포상피-간충질전화(EMT)적작용급궤제.방법 채용불동농도적Erlotinib작용우폐선암세포주H1650,유도기산생내약(H1650ER);면역형광법급Western blot검측내약전후상피형개점단백(E-cadherin)、파형단백(Vimentin)、CD146적표체변화;소간우RNA(siRNA)간우H1650ER세포중CD146적표체,이용Transwell침습소실급세포계수시제합(CCK-8)실험분별평개폐선암세포침습급증식능력,Western blot 검측상관단백적표체변화.결과 획득성Erlotinib내약폐선암세포H1650ER중CD146급Vimentin 단백적표체량분별위0.474±0.013화0.465±0.012,현저고우내약전(P＜0.05);이E-cadherin단백적표체량위0.180±0.011,명현저우내약전(P＜0.05).CD146-siRNA간우H1650ER세포CD146적표체후가명현억제기증식능력,여대조조비교차이유통계학의의(P＜0.05);CD146-siRNA간우후세포침습력역하강(P＜0.05).결론 획득성Erlotinib내약폐선암산생EMT병CD146표체현저상조;억제CD146적표체가하강기침습급증식능력.
Objective To elucidate the impact of CD146 on epithelial-mesenchymal transition (EMT) of acquired resistance to erlotinib in non-small cell lung cancer (NSCLC),thus to reveal the molecular mechanism of CD146 involved in the EMT and in the metastasis of cancer cells.Methods NSCLC cell line H1650 was used to establish the subline resistant to erlotinib (H1650ER).The expression of E-cadherin,Vimentin and CD146 in the resistant cells was examined using immunostaining and Western blotting.CD146 was silenced by RNA interference in H1650ER,and invasion assay was performed to characterize the resistant cells.The effects of CD146 on cell proliferation and invasion were analyzed.Results The quantity of CD146 expression and Vimentin expression in lung adenocarcinoma cancer cell H1650ER were 0.474 ± 0.013 and 0.465 ± 0.012,significantly higher than before resistance (P ＜ 0.05).The E-cadherin protein expression was 0.180 ± 0.011,significantly lower than before resistance (P ＜ 0.05).As compared with the control group,small interfering RNA (siRNA)-mediated down-regulation of CD146 suppressed the proliferation and invasion ability of H1650ER (P ＜ 0.05).Conclnsion CD146 contributes to EMT of acquired resistance to erlotinib in NSCLC in vitro.It suggests that CD146 is a potential molecular target in the treatment of NSCLC.