MiR106a对甲状腺癌细胞增殖、凋亡及侵袭性的影响  

Effects of miR106a on the proliferation, apoptosis and invasion of thyroid cancer cells in vitro

作　　者：沈晨天 邱忠领[1] 魏伟军[1] 宋红俊[1] 罗全勇[1] 

机构地区：[1]上海交通大学附属第六人民医院核医学科,200233

出　　处：《中华核医学与分子影像杂志》2017年第8期486-491,共6页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：国家自然科学基金（81201115）

摘　　要：目的 研究微小RNA（miR）106a 对甲状腺癌细胞增殖、凋亡及侵袭能力的影响.方法 以人未分化甲状腺癌细胞株8505C和PTC细胞CGTH-W3为研究对象,利用慢病毒载体实现细胞内miR106a的过表达及抑制.采用实时荧光定量 PCR（qRT-PCR）及Western blot法检测相关基因的表达,采用MTT法检测上述细胞的增殖活性变化,用流式细胞仪检测细胞凋亡.检测各组细胞含半胱氨酸的天冬氨酸蛋白水解酶（caspase）-9活性,并采用划痕实验及Transwell侵袭小室实验检测细胞侵袭能力.组间数据比较采用两样本t检验、单因素方差分析.结果 MiR106a在8505C细胞中的表达高于CGTH-W3（t=10.28,P〈0.01）.成功构建并分别稳定转染miR106a抑制物基因、无义寡聚核苷酸基因、miR106a基因的细胞株8505C-miR106a（-）、8505C-control、CGTH-W3-miR106a（＋）、CGTH-W3-control.8505C细胞在抑制miR106a后,其增殖活性明显下降,8505C、8505C-control、8505C-miR106a（-）组间差异有统计学意义（F=147.0,P〈0.001）;凋亡比例明显升高,细胞中caspase-9活性明显升高,迁移和侵袭能力明显下降,3组间差异均有统计学意义（F=537.8、804.3、19.2、100.3,均P〈0.01）.CGTH-W3过表达miR106a后,其增殖活性明显升高,CGTH-W3、CGTH-W3-control、CGTH-W3-miR106a（＋）组间差异有统计学意义（F=9.2,P〈0.01）;凋亡比例下降,细胞中caspase-9活性下降,迁移和侵袭能力明显升高（F=12.3、19.6、13.3、622.8,均P〈0.01）.Western blot结果显示,8505C细胞在抑制miR106a后, MEKK2及p-ERK1/2表达明显下降,而CGTH-W3过表达miR106a后,MEKK2及p-ERK1/2表达明显升高.结论 MiR106a在甲状腺癌细胞中发挥着癌基因的作用,即可促进甲状腺癌细胞增殖,增加其侵袭及迁移能力并抑制凋亡.Objective To investigate the effects of microRNA （miR）106a on the proliferation, apoptosis, migration and invasion of thyroid cancer cells in vitro.Methods 8505C and CGTH-W3 cell lines were used in the study.Overexpression and inhibition of miR106a were achieved by transfection of lentiviral vectors.The changes of gene expression were detected by quantitative real-time PCR （qRT-PCR） and Western blot analysis.Cell viability and apoptosis were evaluated by MTT assay and flow cytometry analysis, respectively.The caspase-9 activities in parental CGTH-W3 and 8505C cells and transfected sublines were measured.Wound healing and Transwell invasion assays were performed to determine cell migration and invasion.Two-sample t test and one-way analysis of variance were used to analyze the data.Results The level of miR106a in 8505C was up-regulated when compared to that in CGTH-W3 cells （t=10.28, P〈0.01）.Scrambled control and miR106a（-） were also successfully transfected into cells.Inhibition of miR106a suppressed cell viability, migration and invasion while promoted apoptosis and caspase-9 activity of 8505C cells, with significant differences among 8505C, 8505C-control, 8505C-miR106a（-） cells （F=147.0, 19.2, 100.3, 537.8, 804.3;all P〈0.01）.Overexpression of miR106a promoted cell viability, migration and invasion while inhibited apoptosis and caspase-9 activity of CGTH-W3 cells, with significant differences among CGTH-W3, CGTH-W3-control, CGTH-W3-miR106a（＋） cells（F=9.2, 13.3, 622.8, 12.3, 19.6, all P〈0.01）.In addition, miR106a may up-regulate the expression of MEKK2 and p-ERK1/2.Conclusion Acting as an onco-miR, miR106a might promote the proliferation, migration and invasion of thyroid cancer cells and inhibit their apoptosis in vitro.

关 键 词：甲状腺肿瘤 细胞增殖 细胞凋亡 肿瘤侵润 微RNAS 肿瘤细胞 培养的 

分 类 号：R736.1[医药卫生—肿瘤]