阿司匹林通过调控EMT对结肠癌HT-29细胞迁移和侵袭的影响  

The effects of aspirin on the migration and invasion of colon cancer HT-29 cells by regulating EMT

作　　者：朱蓉[1] 陈曦[1] 俆永素 鲁生念[1] 李伶俐[1] 赵逵[1] ZHU Rong;CHEN Xi;XU Yong - su;LU Sheng - nian;LI Ling - li;ZHAO Kui(Department of Gastroenterology, Affiliated Hospital of Zunyi Medical College, Guizhou Institute of Digestive Disease, Zunyi 563000, Guizhou, Chin)

机构地区：[1]遵义医学院附属医院消化内科贵州省消化病研究所,贵州遵义563000

出　　处：《广东医学》2018年第7期974-978,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81560467);贵州省科学技术基金项目(编号:黔科合J字LKZ[2013]18号);贵州省卫生计生委科学技术基金项目(编号:gzwjkj2017-1-001);遵义医学院博士科研启动基金项目(编号:F-572)

摘　　要：目的探讨非甾体类抗炎药阿司匹林对人结肠癌HT-29细胞株上皮-间充质转变(EMT)、迁移和侵袭能力的影响。方法体外培养HT-29细胞株,使用不同浓度的阿司匹林(0、2.5、5、10、15 mmol/L)干预,采用噻唑蓝(MTT)比色法检测不同浓度阿司匹林对HT-29细胞生长增殖的影响,并计算出半数抑制浓度(IC_(50));分别采用实时定量逆转录聚合酶链反应(real-time PCR)及蛋白印迹(Western blot)法检测阿司匹林IC_(50)干预0、24、48 h后EMT上皮标志物(E-cadherin)和间充质标志物(N-cadherin、Vimentin、Snail1)mRNA以及蛋白质的表达;同时采用细胞划痕及Transwell小室侵袭实验观察阿司匹林对HT-29细胞迁移和侵袭的影响。结果阿司匹林呈时间和剂量依赖性地抑制HT-29细胞生长增殖,IC_(50)为5.086 mmol/L;阿司匹林干预48 h,HT-29细胞上皮标志物E-cadherin mRNA及蛋白质的表达均较对照组上调(P<0.05);间充质标志物N-cadherin、Vimentin及Snail1 mRNA及蛋白质的表达均较对照组下调(P<0.05);阿司匹林干预48 h,HT-29细胞的迁移及侵袭能力均减弱(P<0.05)。结论非甾体类抗炎药阿司匹林通过上调Ecadherin表达,下调N-cadherin、Vimentin、Snail1表达从而抑制HT-29细胞EMT;阿司匹林可通过抑制EMT从而抑制HT-29细胞的迁移及侵袭能力。Objective To investigate the effects of non - steroidal anti - inflammatory drug aspirin on epithelial - mesenchymal transition, migration and invasion capabilities of cultured human colon cancer cell line HT - 29. Methods HT -29 colon cancer cells were cultured in vitro, and different concentrations of aspirin （0, 2.5, 5, 10, 15 mmol/L） were added to the cells. MTr colorimetric assay was used to detect proliferation rates of HT - 29 cells, and the half inhibi- tory concentration （ICs0） was calculated. Real -time quantitative reverse transcription polymerase chain reaction （real - time PCR） was used to detect mRNA expressions of epithelial marker （E - eadherin） and mesenchymal markers （N - cadherin, vimentin and snaill ） after aspirin intervention for 0, 24, 48 hours. Western blotting was used to detect their protein expressions. Cell wound scratch assay was used to observe the effects of aspirin on migration capability of HT - 29 cells, and Transwell chamber invasion assay was used to observe the effects on invasion ability. Results MTT results showed that aspirin inhibited the proliferation of HT - 29 cells in a time - and dose - dependent manner, and the IC50 was 5. 086 mmol/L. Real - time PCR and Western blotting showed that the mRNA and protein expressions of epithelial marker （E- cadherin） were increased （P 〈0. 05 ）, and the mRNA and protein expressions of mesenchymal markers （N- cad- herin, vimentin and snaill ） were decreased （P 〈 0. 05 ）, compared with the control group, after aspirin intervention for 48 hours. Cell wound scratch assay showed that the migration ability of HT - 29 cells was weakened, and Transwell cham- ber invasion assay showed that the invasion ability was weakened, too. （ P 〈 0. 05） Conclusion Non - steroidal anti - inflammatory drug aspirin can inhibit the EMT process through increasing the expression of E - cadherin and decreasing the expressions of N -cadherin, Vimentin and Snaill. Aspirin can inhibit the migration and invasion of HT- 29

关 键 词：阿司匹林 结肠癌 上皮-间充质转变 E-钙黏蛋白 非甾体类抗炎药 

分 类 号：R735.35[医药卫生—肿瘤]