摘要
目的:探讨甘草酸(GA)通过调控miR-142/锌指E盒结合的同源盒蛋白1(ZEB1)分子轴对非小细胞肺癌(NSCLC)HCC827和A549细胞增殖、侵袭和迁移的影响。方法:HCC827和A549细胞培养和转染完成后,分成4组:NC组(未经转染+3mmol/L GA)、miR-142 inhibitor组(敲降miR-142+3 mmol/L GA)、pcDNA3.1-ZEB1组(过表达ZEB1+3 mmol/L GA)和pcDNA3.1-ZEB 1+miR-142mimic组(过表达ZEB1及miR-142+3 mmol/LGA)。采用qPCR检测不同浓度GA处理后HCC827和A549细胞中miR-142的表达水平,WB实验检测HCC827和A549细胞中ZEB1蛋白的表达水平,采用MTT和Transwell检测HCC827和A549细胞的增殖、侵袭和迁移能力,采用双荧光素酶报告基因检测miR-142与ZEB1的靶向关系。结果:GA显著抑制HCC827和A549细胞的增殖、侵袭和迁移,且显著上调miR-142的表达水平(P<0.05或P<0.01);miR-142通过靶向结合ZEB1的3’-UTR区域下调ZEB1的表达水平(P<0.05或P<0.01);进一步实验证实,GA通过上调miR-142抑制ZEB1的表达水平,进而抑制HCC827和A549细胞增殖、侵袭和迁移(P<0.05或P<0.01)。结论:GA能够抑制NSCLC HCC827和A549细胞增殖、侵袭和迁移,其机制为GA通过上调miR-142对ZEB1的抑制作用,从而抑制HCC827和A549细胞的恶性生物学行为。
Objective:To explore the effect of glycyrrhizin(GA) on the proliferation,invasion and migration of non-small cell lung cancer HCC827 and A549 cells via regulating miR-142/ZEB1(Zinc finger E-box-binding homeobox 1) axis.Methods:After being cultured and transfected,HCC827 and A549 cells were divided into 4 groups:NC group(untransfected+3 mmol/L GA),miR-142 inhibitor group(miR-142 knockdown+3 mmol/L GA),pcDNA3.1-ZEB1 group(ZEB1 over-expression+3 mmol/L GA) and pcDNA3.1-ZEB1+miR-142 mimic group(ZEB1 over-expression+miR-142+3 mmol/L GA).qPCR was used to detect the expression level of miR-142 in HCC827 and A549 cells treated with different concentrations of GA.MTT and Transwell assays were used to examine the proliferation,invasion and migration of HCC827 and A549 cells.WB was used to detect the expression level of ZEB1 protein in HCC827 and A549 cells.Dual-luciferase reporter gene assay was used to explore the relationship between miR-142 and ZEB1.Results:GA significantly inhibited the proliferation,invasion and migration of HCC827 and A549 cells,and up-regulated the expression level of miR-142(P<0.05 or P<0.01).Dual-luciferase reporter gene assay showed that miR-142 could targetedly combine with 3’-UTR of ZEB1 and downregulate the expression of ZEB1(P<0.05 or P<0.01).Further experiment validated that GA inhibited ZEB1 expression via up-regulating miR-142,thus suppressed proliferation,invasion and migration of HCC827 and A549 cells(P<0.05 or P<0.01).Conclusion:GA inhibits the proliferation,invasion and migration of NSCLC HCC827 and A549 cells,the mechanism of which is that GA inhibits the malignant biological behavior of NSCLC HCC827 and A549 cells via up-regulating the inhibition effect of miR-142 on ZEB1.
作者
赵润杨
孟泳
王艳梅
侯从岭
ZHAO Runyang;MENG Yong;WANG Yanmei;HOU Congling(Department of Respiratory,the Second Affiliated Hospital of Henan University of Traditional Chinese Medicine&Henan Hospital of Traditional Chinese Medicine,Zhengzhou 450003,Henan,China)
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2019年第12期1337-1344,共8页
Chinese Journal of Cancer Biotherapy
