miR-125a靶向E2F2抑制口腔鳞状细胞癌干细胞干性的初步研究  

Preliminary study of miR-125a targeting E2F2 to inhibit stem cell stemness of oral squamous cell carcinoma

作　　者：陈宏丽 赵洪波[2] 尹刚 CHEN Hong-Li;ZHAO Hong-Bo;YIN Gang(Department of Stomatology,Fenyang College of Shanxi Medical University,Fenyang 032200,China)

机构地区：[1]山西医科大学汾阳学院口腔医学教研室,汾阳032200 [2]山西医科大学附属汾阳医院口腔科,汾阳032200

出　　处：《中国免疫学杂志》2020年第23期2872-2878,共7页Chinese Journal of Immunology

基　　金：山西省卫生计生科研课题(20150183)。

摘　　要：目的:研究miR-125a在口腔鳞状细胞癌(FOSCC)组织中的表达及与OSCC患者临床病例特征的关系,并探讨miR-125a对E2F2的靶向调控作用和对OSCC干细胞(CSCs)干性的影响。方法:qRT-PCR检测miR-125a在40例OSCC组织和20例正常口腔黏膜组织中的表达水平,并分析OSCC组织中miR-125a的表达与临床病例特征的关系。磁珠分选OSCC细胞系Cal-27中的CSCs。Western blot检测OCT4、Nanog、SOX2干性标志蛋白及miR-125a在CSCs中的表达;脂质体分别转染miR-125a mimic和对照NC 48 h后,MTS检测各组CSCs增殖能力,软琼脂克隆检测各组CSCs肿瘤球形成能力,Transwell实验检测各组CSCs转移能力。Targetscan7.1软件及双荧光素酶报告基因试验,研究miR-125a对E2F2基因的靶向作用,qRT-PCR检测miR-125a对CSCs中E2F2 mRNA表达的影响。结果:qRT-PCR结果显示miR-125a在OSCC组织中的表达显著低于正常口腔黏膜组织,其低表达与OSCC肿瘤直径大、TNM分期晚期及淋巴结转移显著相关(P<0.05)。OCT4、Nanog、SOX2干细胞标志物在CSCs中表达显著上调;miR-125a在CSCs中的表达均显著低于OSCC细胞Cal-27和人口腔黏膜角化细胞HOK(P<0.05);转染miR-125a mimic后,CSCs细胞增殖、肿瘤球形成及转移能力均下降(P<0.05)。TargetScan7.1软件和双荧光素酶报告基因试验证实E2F2为miR-125a的靶基因,miR-125a mimic组E2F2 mRNA表达降低(P<0.05)。结论:miR-125a在OSCC组织和细胞系中均低表达,且低表达与肿瘤直径大、TNM分期晚期及淋巴结转移相关,miR-125a可能通过负调控E2F2抑制CSCs的干性。Objective:To study the expression of miR-125a in oral squamous cell carcinoma(OSCC)and relationship between miR-125a and clinical case characteristics of OSCC patients,and to explore the targeted relationship of miR-125a on E2F2 and its effect on stemness of OSCC cancer stem cells(CSCs).Methods:qRT-PCR was used to detect expression levels of miR-125a in 40 OSCC tissues and 20 normal oral mucosa tissues,relationship between miR-125a expression in OSCC tissues and clinical case characteristics was analyzed.Magnetic beads were used to sort CSCs in OSCC cell line Cal-27.Western blot was used to detect expression of OCT4,Nanog,SOX2 stem marker protein and miR-125a in CSCs.After transfected by lipidosome with miR-125a mimic and control NC for 48 h,MTS was used to detect proliferation of CSCs in each group,soft agar was used to detect tumor formation ability of CSCs in each group,Transwell was used to detect transfer ability of CSCs in each group.Targetscan 7.1 software and dual luciferase reporter gene assay were used to study targeting effect of miR-125a on E2F2 gene,and qRT-PCR was used to detect effect of miR-125a on E2F2 mRNA expression in CSCs.Results:Results of qRT-PCR showed that expression of miR-125a in OSCC tissues was significantly lower than that in normal oral mucosa,and it′s low expression was significantly correlated with OSCC large tumor diameter,advanced TNM stage and lymph node metastasis(P<0.05).Expressions of OCT4,Nanog and SOX2 stem cell markers were significantly up-regulated in CSCs,expression of miR-125a in CSCs was significantly lower than that in OSCC cells Cal-27 and human oral mucosal keratinocytes HOK(P<0.05).After transfected miR-125a mimic,the proliferation,tumor ball formation and metastasis ability of CSCs were decreased(P<0.05).TargetScan7.1 software and dual luciferase reporter gene assay confirmed that E2F2 was a target gene of miR-125a,and expression of E2F2 mRNA in miR-125a mimic group was decreased(P<0.05).Conclusion:miR-125a is lowly expressed in OSCC tissues and cell lines,its

关 键 词：OSCC miR-125a E2F2 干性 

分 类 号：R739.8[医药卫生—肿瘤]