circRNA MYO1C对肺癌细胞自噬和肿瘤干细胞分化的影响

The effects of circRNA MYO1C on lung cancer cell autophagy and cancer stem cell differentiation

目的探讨环状RNA(circRNA)MYO1C对非小细胞肺癌(NSCLC)细胞自噬和肿瘤干细胞(Cancer stem cells,CSCs)分化的影响。方法通过RT-qPCR检测正常肺上皮细胞和NSCLC A549细胞中circRNA MYO1C水平。将A549分为对照组、NC组和siMYO1C组,通过转染si-circRNA MYO1C质粒干扰circRNA MYO1C。用CCK-8法和Transwell实验检测细胞生长活力和迁移能力。通过Western blot检测CSCs分化标志蛋白(OCT4、SOX2和NANOG)以及自噬标志蛋白(P62和IC3II/LC3I)的表达水平。结果相对正常肺上皮细胞,circRNA MYO1C在A549细胞中表达明显上调(0.99±0.06 vs.4.87±0.88,P=0.002)。与NC组比较,siMYO1C组细胞中circRNA MYO1C水平、细胞活力、迁移、OCT4、SOX2、NANOG和IC3II/LC3I蛋白水平显著降低,而P62蛋白水平显著升高(P<0.001)。结论下调circRNA MYO1C能够抑制NSLCLC细胞的CSCs分化和自噬相关蛋白表达,并抑制肺癌细胞迁移。

Objective The aim of this study was to investigate the effects of circRNA MYO1C on autophagy and cancer stem cells(CSCs)differentiation of non-small cell lung cancer(NSCLC).Methods The levels of circRNA MYO1C in normal lung epithelial cells and NSCLC A549 cells were detected by RT-qPCR.A549 cells were divided into the control,NC and siMYO1C groups,and circRNA MYO1C was interfered by transfection with si-circRNA MYO1C plasmids.The cell growth viability and migration ability were detected by CCK-8 and Transwell assays.The expression of CSCs differentiation marker proteins(OCT4,SOX2 and NANOG)and autophagy marker proteins(P62 and IC3II/LC3I)was detected by Western blot.Results Compared with normal lung epithelial cells,the expression of circRNA MYO1C was significantly up-regulated in A549 cells(0.99±0.06 vs.4.87±0.88,P=0.002).Compared with the NC group,the level of circRNA MYO1C,cell viability,cell migration,the levels of OCT4,SOX2,NANOG and IC3II/LC3I proteins in the siMYO1C group were significantly reduced,the level of P62 protein was significantly increased(P<0.001).Conclusion Down-regulating circRNA MYO1C can inhibit CSCs differentiation and autophagy-related protein expression of NSLCLC cells,and inhibit lung cancer cell migration.