MiR-579-3p靶向肌动蛋白相关蛋白3B调控口腔鳞状细胞癌细胞增殖和凋亡的机制研究  

Mechanism of MiR-579-3p Regulating Proliferation and Apoptosis of Oral Squamous Cell Carcinoma Cells by Targeting ACTR3B

作　　者：高功杰 王海业 李晓彦[3] GAO Gongjie;WANG Haiye;LI Xiaoyan(Zhide Stomatology Hospital of Zhengzhou,Zhengzhou 450052;Department of Stomatology,the First Hospital,Zhengzhou University,Zhengzhou 450033;Department of General Surgery,Zhengzhou People's Hospital East District Branch,Zhengzhou 450003,Henan Province,China)

机构地区：[1]郑州植得口腔医院,河南郑州450052 [2]郑州大学第一附属医院口腔科,河南郑州450033 [3]郑州人民医院郑东院区普外科,河南郑州450003

出　　处：《口腔颌面外科杂志》2020年第3期156-162,共7页Journal of Oral and Maxillofacial Surgery

摘　　要：目的:探讨miR-579-3p对口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)细胞增殖和凋亡的影响及作用机制。方法:培养正常人口腔角质形成细胞(normal human oral keratinocyte,NHOK)和OSCC细胞系CAL27、CAL33、SCC15,实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测细胞中miR-579-3p和肌动蛋白相关蛋白3B(actin-related protein 3B,ACTR3B)信使RNA(mRNA)的表达水平,蛋白质印迹法(western blot)检测ACTR3B蛋白表达水平。以CAL33细胞为研究对象,构建过表达miR-579-3p或沉默ACTR3B表达的CAL33细胞,四甲基噻唑蓝染色法(methylthiazoletrazolium,MTT)检测细胞增殖,流式细胞仪检测细胞凋亡,western blot检测细胞周期蛋白D1(CyclinD1)和活化的半胱天冬酶-3(C-caspase-3)蛋白水平。Starbase生物信息学软件预测ACTR3B可能是miR-579-3p的靶基因,双荧光素酶报告基因实验验证miR-579-3p和ACTR3B调控关系。结果:与NHOK细胞比较,OSCC细胞系CAL27、CAL33和SCC15中miR-579-3p表达水平降低(P<0.05),ACTR3B mRNA和蛋白表达水平升高(P<0.05)。过表达miR-579-3p或沉默ACTR3B表达均可降低CAL33细胞存活率和CyclinD1蛋白表达水平(P<0.05),提高CAL33细胞凋亡率和C-caspase-3蛋白表达水平(P<0.05)。miR-579-3p靶向负调控ACTR3B表达。过表达ACTR3B降低了过表达miR-579-3p对CAL33细胞存活率、凋亡率及CyclinD1和C-caspase-3蛋白表达的影响。结论:过表达miR-579-3p可能通过下调ACTR3B表达抑制OSCC细胞增殖,并诱导细胞凋亡。Objective:To investigate the effect and mechanism of miR-579-3p on proliferation and apoptosis of oral squamous cell carcinoma(OSCC)cells.Methods:The normal human oral keratinocytes(NHOK)and oral squamous carcinoma cell lines CAL27,CAL33 and SCC15 were cultured.The expression levels of miR-579-3p and actin-related protein 3B(ACTR3B)mRNA were detected by quantitative real-time PCR(qRT-PCR).The expression level of ACTR3B protein was detected by western blot.Taking CAL33 cells as the research object,CAL33 cells with miR-579-3p overexpressed or ACTR3B expression silenced were constructed.Then cell proliferation was detected by methylthiazoletrazolium(MTT)assay,apoptosis was detected by flow cytometry,and the expression levels of CyclinD1 and C-caspase-3 protein were detected by western blot.Starbase Bioinformatics software predicts that ACTR3B may be the target gene of miR-579-3p,and the dual luciferase reporter gene assay validates the regulatory relationship between miR-579-3p and ACTR3B.Results:Compared with NHOK cells,the expression levels of miR-579-3p in oral squamous cell carcinoma cell lines CAL27,CAL33 and SCC15 were decreased(P<0.05),and the expression levels of ACTR3B mRNA and protein were increased(P<0.05).Overexpression of miR-579-3p or silencing ACTR3B could decrease the survival rate of CAL33 cells and the expression level of CyclinD1 protein(P<0.05),and increase the apoptosis rate of CAL33 cells and the expression level of C-caspase-3 protein(P<0.05).MiR-579-3p targets negative regulation of ACTR3B expression.Overexpression of ACTR3B reduced the effect of overexpression of miR-579-3p on CAL33 cell viability,apoptotic rate,and the expression level of CyclinD1 and C-caspase-3 protein.Conclusion:Overexpression of miR-579-3p may inhibit the proliferation of oral squamous carcinoma cells and induce apoptosis by down-regulating the expression of ACTR3B.

关 键 词：miR-579-3p 肌动蛋白相关蛋白3B 口腔鳞状细胞癌 细胞增殖 凋亡 

分 类 号：R782[医药卫生—口腔医学]