Mir-30a抑制自噬及上皮细胞间质化增强肺腺癌细胞对克唑替尼的敏感性

目的探索上调肺腺癌细胞中Mir-30a的表达水平能否增强细胞对克唑替尼的敏感性,观察细胞增殖率的变化及克唑替尼IC50等的变化,并探索相关的机制。方法使用Lipofectamine2000携带Mir-30a对H3122细胞进行瞬时转染,并与克唑替尼共同作用。将肺腺癌H3122细胞分为对照组、克唑替尼与联合组。MTT法检测各组细胞增殖率变化,Transwell检测细胞侵袭性。Western blot检测ALK、c-MET、Beclin-1、E-cadherin及Vimentin蛋白的表达。结果在H3122细胞中Mir-30a转染与克唑替尼共同作用较克唑替尼单独作用有更显著的细胞杀伤作用。转染后的细胞较单独应用克唑替尼组侵袭力减弱。转染与克唑替尼联合组Beclin1、ALK、c-MET及Vimentin的蛋白的表达有明显下降,E-cadherin稍有增强。结论 Mir-30a增强肺腺癌细胞对克唑替尼的敏感性,可能与过量表达的Mir-30a抑制癌细胞的自噬及上皮细胞间质化相关。

Mir-30a enhances the sensitivity of lung adencarcinoma cells to Crizotinib by inhibiting autophagy and epithelial mesenchymal transition

Objective To investigate if upregulating Mir-30 a expression in human lung adencarcinoma cells（H3122 cells） can enhance the cells sensitivity to Crizotinib, to explore the related mechanism. Methods Lipofectamine2000 carrying Mir-30 a was transfected into H3122 cells. H3122 cells were divided into control group, Crizotinib group and Mir-30 a trasfection ＋ Crizotinib group. The proliferation and invasiveness of the cells were detected by MTT and Transwell respectively. The expression of ALK, c-MET, Beclin-1, E-cadherin and Vimentin proteins was detected by Western blot. Results The inhibition effect on H3122 cells was much more strong in Mir-30 a trasfection ＋ Crizotinib group than in Crizotinib group, but the invasive ability of the cells was decreased in Mir-30 a trasfection ＋ Crizotinib group. The expression levels of ALK, c-MET, Beclin-1 and Vimentin proteins were decreased, but increased for E-cadherin in Mir-30 a trasfection ＋ Crizotinib group than in Crizotinib group. Conclusion Mir-30 a can enhance the sensitivity of H3122 cells to Crizotinib, which might be related to the overexpression of Mir-30 a inhibiting the autophagy and epithelial mesenchymal transition.