miR-206调控小细胞肺癌干细胞EVI1基因表达及细胞生物学行为  

miR-206 regulates EVI1 gene expression and cell biological behavior in stem cells of small cell lung cancer

作　　者：王继芳 鲍祯 乔亚红[1] Wang Jifang;Bao Zhen;Qiao Yahong(Henan Provincial Chest Hospital,Zhengzhou 450000,Henan Province,China)

机构地区：[1]河南省胸科医院,河南省郑州市450000

出　　处：《中国组织工程研究》2022年第7期1027-1031,共5页Chinese Journal of Tissue Engineering Research

基　　金：河南省医学科技攻关计划项目(LHGJ20190727),项目负责人:王继芳。

摘　　要：背景:通过对小细胞肺癌体外分选鉴定,可以发现肿瘤干细胞的存在是促使肿瘤细胞恶性增殖的重要因素。肿瘤的发生与细胞内多种分子的异常表达以及靶向基因的调控障碍密切相关。前期研究发现,EVI1具备原癌基因特性,在肺癌中其表达上调。miR-206可靶向调控EVI1基因和蛋白的表达。目的:探讨miR-206靶向调控EVI1转录基因的表达对小细胞肺癌干细胞增殖和分裂周期的影响。方法:首先采用miR-206转染实验验证靶向调控EVI1基因,实验分3组:miR-206 mimics组、miR-206 inhibitor组和miR-NC组,分别转染小细胞肺癌干细胞,Western blot法检测细胞中EVI1蛋白的表达。然后采用慢病毒干扰法下调小细胞肺癌干细胞中miR-206表达验证细胞的信号转导通路和生物学行为,实验分3组:pLB-miR-206组、空载体组和对照组,RT-PCR法检测细胞中miR-206和EVI1 mRNA表达水平,Western blot法检测p-Akt和p-JNK蛋白表达,MTT法检测细胞增殖活性,流式细胞术检测细胞周期中分裂细胞和凋亡细胞的百分比。结果与结论:①miR-206 mimics组Evi1蛋白水平显著高于miR-NC组(P<0.05),miR-206 inhibitor组Evi1蛋白水平显著低于miR-NC组(P<0.05),提示miR-206可能靶向调控小细胞肺癌干细胞EVI1基因的表达;②经miR-206慢病毒转染后,pLB-miR-206组miR-206和EVI1 mRNA表达水平明显低于空载体组和对照组,p-Akt和p-JNK蛋白表达水平下降,细胞增殖率减少,分裂细胞比例下降(P<0.05),提示miR-206可能通过靶向调控小细胞肺癌干细胞中EVI1基因的表达和Akt/JNK信号通路的激活,对细胞增殖和分裂周期产生影响。BACKGROUND:Through sorting and identification of small cell lung cancer in vitro,the existence of tumor stem cells has been shown to be an important factor to promote the malignant proliferation of tumor cells.Tumorigenesis is closely related to the abnormal expression of various molecules in cells and the regulation of targeted genes.Previous studies have found that EVI1 has the characteristics of proto oncogene and can be up-regulated in lung cancer.miRNA-206 can target the expression of EVI1 gene and protein.OBJECTIVE:To investigate the influence of miRNA-206 on cell proliferation and division cycle by targeted regulating the expression of EVI1 transcription gene in stem cells of small cell lung cancer.METHODS:Firstly,miRNA-206 transfection experiment was used to verify the targeted regulation of EVI1 gene.The experiment was divided into three groups:miRNA-206 mimics,miR-206 inhibitor,and miR-NC.The expression of EVI1 protein in the stem cells was detected by western blot assay.miRNA-206 lentivirus transfection experiment was used to verify the signal transduction pathway and biological behavior of the cells.The experiment was divided into three groups:pLB-miR-206 group,empty vector group,and control group.The expression levels of miR-206 and EVI1 mRNA in stem cells were detected by RT-PCR.The expression levels of p-Akt and p-JNK protein were detected by western blot assay.The cell proliferation rate was detected by MTT method.Percentage of mitotic and apoptotic cells in cell cycle was detected by flow cytometry.RESULTS AND CONCLUSION:(1)The Evi1 protein level in the miR-206 mimics group was significantly higher than that in the miR-NC group(P<0.05),and the Evi1 protein level in the miR-206 inhibitor group was significantly lower than that in the miR-NC group(P<0.05).It was suggested that miR-206 may target the expression of EVI1 gene in stem cells.(2)After transfection with miRNA-206 lentivirus,the expression levels of miR-206 and EVI1 mRNA in pLB-miR-206 group were significantly lower than those in the empty

关 键 词：干细胞 肿瘤干细胞 肺癌干细胞 小细胞肺癌 miR-206 EVI1基因 信号通路 增殖 细胞周期 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学]