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[目的]研究miR-21是否可以调控上皮—间质转化(EMT)而参与肺癌获得性耐药。[方法]使用HCC827细胞(EGFR基因19外显子缺少的肺腺癌细胞株),在此细胞的基础上培养吉非替尼耐药细胞株HCC827/GR。检测耐药细胞株中miR-21的表达,同时观察耐药细胞的形态变化,及在耐药细胞株中抑制miR-21表达后检测耐药细胞株的侵袭能力及EMT相关蛋白E-cadherin和Vimentin的表达。[结果]与敏感细胞株HCC827相比,HCC827/GR细胞株对吉非替尼的耐药倍数约为100倍;同时在耐药细胞株中我们发现miR-21较敏感株表达增高约5.3倍,同时细胞形态发生了明显变化。通过检测EMT相关蛋白,我们发现耐药细胞株中间质相关蛋白Vimentin明显高表达,而上皮相关蛋白E-cadherin明显低表达,当miR-21被抑制后,耐药细胞株侵袭能力下降,同时Vimentin表达量下降,而E-cadherin表达量增高。[结论]miR-21可能通过促进EMT参与肺癌EGFR-TKI的获得性耐药。
[Objective] To investigate whether miR-21 can regulate epithelial-mesenchymal transition (EMT) and participate in acquired drug resistance of lung cancer. [Method] HCC827 cells (lung adenocarcinoma cell line lacking EGFR gene exon 19) were used to culture gefitinib resistant cell line HCC827 / GR on the basis of this cell line. To detect the expression of miR-21 in drug-resistant cell lines, observe the morphological changes of drug-resistant cells, detect the invasion ability of drug-resistant cell lines after inhibiting miR-21 expression in drug-resistant cell lines and the expression of EMT-related protein E-cadherin and Vimentin expression. [Results] Compared with the sensitive cell line HCC827, the resistance multiple of HCC827 / GR cell line to gefitinib was about 100-fold. In the resistant cell line, we found that the expression of miR-21 was increased about 5.3 times At the same time, the cell morphology changed significantly. Through the detection of EMT-related proteins, we found that Vimentin was highly expressed in drug-resistant cell lines, whereas E-cadherin was significantly down-regulated. When miR-21 was inhibited, the invasion ability of drug-resistant cell lines decreased Vimentin expression decreased, while E-cadherin expression increased. [Conclusion] miR-21 may be involved in acquired resistance of EGFR-TKI in lung cancer by promoting EMT.