摘要
目的:分析miRNA-145-5p调控形成素2(formin-like 2,FMNL2)基因对口腔鳞癌干细胞增殖、迁移的影响。方法:按照脂质体2000说明书对细胞进行转染miRNA-145-5p inhibitor及miRNA-145-5p mimics,按照实验设计,将其分为空白组、沉默组(miRNA-145-5p inhibitor)及过表达组(miRNA-145-5p mimics)。荧光定量PCR法检测miRNA-145-5p、FMNL2表达量,MTT检测细胞增殖能力,流式细胞仪检测细胞凋亡能力,细胞划痕实验检测细胞迁移能力,采用Western blot法检测各组细胞中Wnt/β-catenin信号通路蛋白表达量。结果:过表达组miRNA-145-5p、Bax蛋白表达量,凋亡率,细胞G0/G1比例高于沉默组,具有统计学差异(P<0.05);过表达组口腔鳞癌干细胞增殖率,FMNL2表达量、MMP-13、β-catenin、Bcl-2、APC蛋白表达量低于沉默组,具有统计学差异(P<0.05);过表达组口腔鳞癌干细胞迁移能力弱于沉默组,具有统计学差异(P<0.05)。结论:miRNA-145-5p通过靶向调控FMNL2,作用于Wnt/β-catenin信号通路调控口腔鳞癌干细胞,进而抑制细胞增殖、迁移。
Objective:To analyze the effect of miRNA-145-5p regulating homologous morphogen-like protein 2(FMNL2)gene on the proliferation and migration of oral squamous cell carcinoma stem cells.Methods:Transfect cells with miRNA-145-5p inhibitor and miRNA-145-5p mimics according to the liposome 2000 instructions,and divide them into blank group,silent group(miRNA-145-5p inhibitor)and overexpression group according to the experimental design(Mi RNA-145-5p mimics).Fluorescence quantitative PCR method was used to detect the expression of miRNA-145-5p and FMNL2,MTT to detect cell proliferation,flow cytometry to detect cell apoptosis,cell scratch test to detect cell migration ability,Western blot to detect Wnt in each group of cells/β-catenin signaling pathway protein expression.Results:The overexpression group miRNA-145-5p,Bax protein expression,apoptosis rate,cell G0/G1 ratio were higher than the silent group,with statistical differences(P<0.05);overexpression group oral squamous cell carcinoma stem cell proliferation rate,FMNL2 expression The protein expression levels of MMP-13,β-catenin,Bcl-2,and APC were lower than those in the silent group,with statistical differences(P<0.05);the migration ability of oral squamous cell carcinoma stem cells in the overexpression group was weaker than that in the silent group,with statistical significance Difference(P<0.05).Conclusion:By targeting FMNL2,mirna-145-5p acts on Wnt/β-catenin signaling pathway to regulate the proliferation and migration of OSCC stem cells.
作者
谭力芯
叶艳艳
熊宇
税桦桦
黄文静
丁大莲
曾宁碧
赵希
TAN Li-xin;YE Yan-yan;XIONG Yu;SHUIHua-hua;HUANG Wen-jing;DING Da-lian;ZENG Ning-bi;ZHAO Xi(Department of stomatology,First Affiliated Hospital of Military Medical University,Chongqing,400038,China)
出处
《现代生物医学进展》
CAS
2021年第12期2221-2225,共5页
Progress in Modern Biomedicine
基金
教育部留学回国人员科研启动基金项目(HG2015-002)
院级军科基金项目(SWH2016JSTSYB-40N)。
