体外RNA干扰RhoA基因对舌癌细胞侵袭的影响
Effects of RhoA gene silencing by RNA interference on invasion of tongue carcinoma
作者：陈正岗 唐永平 童磊 王莹 周元 王奇民 韩金宏 何宗轩 廖奕翔 樊兵 邹荣海 张健 孙晓峰 严国鑫
Author：Chen Zhenggang, Tang Yongping, Tong Lei, Wang Ying, Zhou Yuan, Wang Qimin, Han Jinhong, He Zongxuan, Liao Yixiang, Fan Bing, Zou Ronghai, Zhang Jian, Sun Xiaofeng, Yan Guoxin.
收稿日期：2015-09-16 年卷（期）页码：2016,34(2):183-183-188
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：RhoA基因,半乳糖凝集素-3,基质金属蛋白酶-9,RNA干扰,舌癌,侵袭,
Key words：RhoA gene,galectin-3,matrix metalloproteinase-9,RNA interference,tongue carcinoma,invasion,
基金项目：
Natural Science Foundation of China (81372908); Science and Technology Developing Major Project of Nanjing Medical University (2012NJMU248); Science and Technology Planning Project of Qingdao Municipal Health and Family Planning Commission (2014-WJZD009, 2013-WSZD011).
中文摘要
目的 应用RNA干扰技术抑制人舌癌细胞株Tca8113和SCC-4中RhoA基因的表达，探讨RhoA基因表达下调对舌癌细胞侵袭的影响。方法 登录Genbank数据库确定人RhoA基因序列，针对RhoA的基因序列设计短链RNA，利用Lipofectamine2000介导法将RhoA-siRNA转染至舌癌Tca8113和SCC-4细胞。采用实时荧光定量聚合酶链反应检测转染后的舌癌细胞中RhoA mRNA的表达，蛋白质印迹法检测RhoA、半乳糖凝集素-3（galectin-3）和基质金属蛋白酶-9（MMP-9）蛋白的表达，Transwell小室检测细胞的侵袭能力。结果 舌癌细胞转染RhoA-siRNA后，RhoA的基因和蛋白表达下降，galectin-3和MMP-9蛋白表达下降，细胞体外侵袭能力降低。结论 RhoA-siRNA可以有效地抑制舌癌Tca8113和SCC-4细胞中RhoA的表达，通过降低galectin-3和MMP-9的表达从而降低细胞的侵袭能力。RhoA在舌癌的侵袭和转移中可能发挥重要的作用。
英文摘要
Objective To study the effects of RhoA down-regulation by RNA interference on the invasion of tongue carcinoma Tca8113 and SCC-4.Methods Determination of the human RhoA sequence as well as the design and construction of a short specific small interfering RNAs (siRNA) were performed. The siRNA of RhoA gene was transfected into human tongue squamous cell carcinoma Tca8113 and SCC-4 cells line by Lipofectamine 2000. Quantitative real-time polymerase chain reaction was used to examine the mRNA expression levels of RhoA. Protein expressions of mRNA, galectin-3, and matrix metalloproteinase (MMP)-9 were evaluated by Western blot. Transwell invasion assay was performed to assess the invasion ability of tongue carcinoma.Results RhoA expressions in Tca8113 and SCC-4 cells were reduced significantly after transfection of RhoA-siRNA. Protein levels of galectin-3 and MMP-9 were also down-regulated significantly. Invasion ability was inhibited as well.Conclusion RhoA-siRNA can effectively inhibit RhoA expression in Tca8113 and SCC-4 cells. The invasion ability of tongue carcinoma cells decreased with down-regulation of the protein expressions of galectin-3 and MMP-9, indicating that RhoA-siRNA can inhibit invasion of tongue carcinoma. Results show that RhoA may play an important role in the processes of invasion and metastasis of tongue carcinoma.
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