EMT参与人非小细胞肺癌A549细胞多西紫杉醇耐药性的发生

Epithelial-mesenchymal transition contributes to human non-small lung cancer cell line A549 docetaxel resistance

目的:建立耐多西紫杉醇的人非小细胞肺癌细胞株A549/Docetaxel,并初步探索其与上皮间质转化(EMT)的关系。方法:用抗肿瘤药物多西紫杉醇体外以浓度递增和短时反复作用的方法诱导和筛选人非小细胞肺癌A549细胞耐多西紫杉醇的细胞株;用MTT的方法检测细胞耐药指数及生长曲线,流式细胞仪检测细胞周期,RT-PCR和Western blot检测EMT相关蛋白fibronectin、vimentin和E-cadherin的表达。结果:A549/Docetaxel对多西紫杉醇和紫杉醇的耐药指数分别为8.91和2.82;耐药细胞生长缓慢,倍增时间延长,S期细胞增多,G2期减少。549/Docetaxel与亲本A549细胞相比,上皮细胞标志分子E-cadherin的表达减少,而间质细胞标志分子fibronectin和vimentin的表达增加。结论:成功建立了一株耐多西紫杉醇的细胞株A549/Docetaxel,耐药的发生可能与EMT相关。

Objective： To establish a new docetaxel resistant human non-small lung cancer cell line（ A549 /Docetaxel） and investigate its association with epithelial-mesenchymal transition（ EMT）. Methods： Docetaxel resistant human non-small lung cancer cell line（ A549 / Docetaxel） was developed by intermittent exposure to gradually increasing concentration of docetaxel from the parent cell line A549 in vitro. The resistance index（ RI） of A549 / Docetaxel to anticancer agents and its growth rate were evaluated by MTT assay. The distribution of its cell cycle was detected by flow cytometry. The expression of fibronectin,vimentin and E-cadherin were detected by RT-PCR and Western blot. Results ： The RI of A549 / Docetaxel cells to docetaxel and paclitaxel were respectively 8. 91 and2. 82. Comparing with the parent cells,the multiplication time of A549 / Docetaxel cells was prolonged. The proportion of cells in G2-phase decreased while those in S-phase increased,and the morphology of the resistant cells was changed obviously. Furthermore,we found the expression of fibronectin and vimentin in resistant cells were significantly higher than that in their parent cells while E-cadherin was much less expressed. Conclusion： We established a typical docetaxel resistant human non-small lung cancer cell line（ A549 / Docetaxel）,and it was suggested that the drug resistance of A549 / Docetaxel was related to EMT.