摘要
目的：观察慢病毒表达载体介导微小核糖核酸－34a（ miR－34a）体外转染口腔癌细胞后对细胞增殖及侵袭的影响，探讨miRNA－34a在口腔癌发展中可能的作用。方法运用实时荧光定量PCR（ RT－PCR）检测口腔癌组织、正常组织样本和口腔癌细胞系中的miR－34aRNA的表达情况，体外培养口腔癌细胞，分别对miR－34a重组慢病毒组和空白对照病毒组转染Tca8113和SCC－25细胞。采用甲基噻唑基四唑（ MTT）法、克隆形成实验法检测转染目的基因细胞增殖活性的变化；采用流式细胞仪检测过表达外源性miR－34a后细胞周期分布的变化情况；采用Transwell小室实验分析转染后Tca8113和SCC－25细胞侵袭和迁移能力的改变。结果与正常组织相比，口腔癌组织中miR－34a的表达显著下调，MTT法和克隆形成率分析显示转染miR－34a基因重组慢病毒转染组与空载体慢病毒转染组比较，细胞增殖明显受到明显抑制（ P＜0．05）；转染后细胞周期结果显示miR－34a对细胞的生长抑制作用部分是通过诱导细胞在G1期出现停滞而实现的。 Transwell小室显示与空载体慢病毒转染组相比，转染miR－34a基因重组慢病毒转染组细胞的迁移力和侵袭力也明显受到抑制（P＜0．05）。结论口腔癌细胞过表达外源性miR－34a后，细胞增殖和侵袭能力明显降低，miR－34a有望在口腔癌治疗中成为新的靶点。
Objective To investigate the transfection of miRNA-34a mediated by lentiviral vectors and its effects on the proliferation and invasion of oral cancer cells, as well as the role of miRNA-34a in the development of oral canc-er.Methods The expression of miRNA-34a was measured in oral cancer cell lines, oral cancer tissues and normal tis-sues by quantitative real-time PCR.Then, oral cancer Tca8113 and SCC-25 cells were transfected with miR-34a or blank control, respectively.The proliferation of these cells was measured by MTT assay and the clony formation assay. The cell cycle was determined by flow cytometry （ FCM） .The metastasis of these cells was detected by Transwell assay. Results Compared with normal tissues, the expression of miRNA-34a were down regulated in oral cancer tissues.Ac-cording to MTT assay and clony formation assay, Tca8113 and SCC-25 cells transfected with LV-miR-34a showed significantly reduced proliferation than the control （P〈0.05）.Such inhibitory effects could be partly attributed to miR-34a induced cell cycle arrest at phase G1 .Moreover, miR-34a over-expression could also remarkably blocked the me-tastasis of oral cancer cells in comparison with the control （P〈0.05）.Conclusions Over expression of miRNA-34a can decrease the proliferation and invasion of oral cancer cells, indicating the potential target of miRNA-34a in the treatment of oral cancer.
出处
《徐州医学院学报》
CAS
2015年第12期912-916,共5页
Acta Academiae Medicinae Xuzhou
基金
国家自然科学基金（31060167,81160242）
江苏省高校“青蓝工程”项目资助
徐州医学院优秀人才科研基金资助
