摘要
目的探讨微小RNA-214(miR-214)对喉鳞状细胞癌(LSCC)细胞增殖和凋亡的影响及可能的分子机制。方法采用实时荧光定量PCR(RT-qPCR)技术检测人LSCC细胞Hep-2、FD-LSC-1、TU177和正常永生化表皮细胞Hacat中的miR-214。将miR-214模拟物(miR-214 mimics)和模拟物阴性对照(miR-NC)转染至TU177细胞,分别记作miR-214 mimics组和miR-NC组,采用RT-qPCR技术检测各组细胞中的miR-214,采用CCK-8实验和平板克隆形成实验分别检测各组细胞增殖活性和克隆形成能力,采用流式细胞术测算各组细胞凋亡率,采用Westernblotting法检测各组细胞中的Bcl-2、Bax和Livin蛋白。采用双荧光素酶报告基因实验检测miR-214和Livin的靶向调控关系。结果Hep-2、FD-LSC-1、TU177细胞中miR-214的相对表达量低于Hacat细胞(P均<0.05)。与miR-NC组相比,miR-214 mimics组TU177细胞增殖活性降低,克隆细胞数减少,凋亡率升高,Livin和Bcl-2蛋白表达降低,Bax蛋白表达升高(P均<0.05)。双荧光素酶报告基因实验显示,miR-214能靶向结合Livin的3'UTR区(P<0.05)。结论miR-214在LSCC细胞中呈低表达;在体外,miR-214能够抑制Livin表达从而抑制LSCC细胞的增殖并促进其凋亡。
Objective To investigate the effects of microRNA-214(miR-214)on proliferation and apoptosis of laryngeal squamous cell carcinoma(LSCC)cells and their possible molecular mechanism.Methods Real-time quantitative PCR(RT-qPCR)was used to detect the expression of miR-214 in human LSCC cells Hep-2,FD-LSC-1,TU177 and normal immortalized epidermal cells Hacat.The miR-214 mimics and mimic negative control(miR-NC)were transfected into TU177 cells,which were recorded as the miR-214 mimics group and miR-NC group,respectively.RT-qPCR was used to detect the expression level of miR-214 in cells of each group,CCK-8 experiment and plate clone formation experiment were used to detect cell proliferation activity and clone formation ability of each group,flow cytometry was used to detect the apoptosis rate of each group,Western blotting was used to detect the expression levels of Bcl-2,Bax and Livin proteins in cells of each group,and the dual luciferase experiment was used to detect the targeted regulation relationship between miR-214 and Livin.Results The expression levels of miR-214 in Hep-2,FD-LSC-1 and TU177 cells was significantly lower than that in Hacat cells(all P<0.05).Compared with the miR-NC group,the proliferation activity of TU177 cells in the miR-214 mimics group was significantly reduced,the number of cloned cells was significantly reduced,and the apoptotic rate increased significantly,the expression levels of Livin and Bcl-2 protein decreased significantly,and the expression level of Bax protein increased(all P<0.05).The dual luciferase experiment showed that miR-214 could target the 3'UTR region of Livin(P<0.05).Conclusions The miR-214 is low expressed in LSCC cells.In vitro,miR-214 can inhibit the expression of Livin to inhibit the proliferation of LSCC cells and promote their apoptosis.
作者
罗德艳
冯俊
彭涛
唐一萍
杨久梅
LUO Deyan;FENG Jun;PENG Tao;TANG Yiping;YANG Jiumei(Department of Otolaryngology,The Second Clinical Medical College of North Sichuan Medical College,Nanchong 637000,China)
出处
《山东医药》
CAS
2021年第28期5-8,共4页
Shandong Medical Journal
基金
四川省科技厅重点研发项目(2018FZ0116)
四川省教育厅自然科学重点项目(18ZA203)
南充市科技局应用技术研究与开发基金项目(16YFZJ0021)。
