circRNA MYO1C对肺癌细胞自噬和肿瘤干细胞分化的影响

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。

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

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.