体外RNA干扰RhoA基因对舌癌细胞侵袭的影响  

Effects of Rho A gene silencing by RNA interference on invasion of tongue carcinoma

作　　者：陈正岗[1,2] 唐永平[1] 童磊[1] 王莹[3] 周元[3] 王奇民[1] 韩金宏[1] 何宗轩 廖奕翔[1] 樊兵[4] 邹荣海[4] 张健[4] 孙晓峰[4] 严国鑫[4] 

机构地区：[1]青岛大学医学院附属青岛市市立医院口腔医学中心,青岛266071 [2]上海交通大学医学院附属第九人民医院口腔颌面外科,上海200011 [3]潍坊医学院口腔医学院,潍坊261021 [4]无锡市第二人民医院口腔科,无锡214002

出　　处：《华西口腔医学杂志》2016年第2期183-188,共6页West China Journal of Stomatology

基　　金：国家自然科学基金(81372908);南京医科大学科技发展基金重点项目(2012NJMU248);青岛市卫计委计划项目(2014-WJZD009;2013-WSZD011)~~

摘　　要：目的应用RNA干扰技术抑制人舌癌细胞株Tca8113和SCC-4中Rho A基因的表达,探讨Rho A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Rho A基因序列,针对Rho A的基因序列设计短链RNA,利用Lipofectamine2000介导法将Rho A-si RNA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Rho A m RNA的表达,蛋白质印迹法检测Rho A、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Rho A-si RNA后,Rho A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 Rho A-si RNA可以有效地抑制舌癌Tca8113和SCC-4细胞中Rho A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。Rho A在舌癌的侵袭和转移中可能发挥重要的作用。Objective To study the effects of Rho A down-regulation by RNA interference on the invasion of tongue carcinoma Tca8113 and SCC-4. Methods Determination of the human Rho A sequence as well as the design and construction of a short specific small interfering RNAs（si RNA） were performed. The si RNA of Rho A 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 m RNA expression levels of Rho A. Protein expressions of m RNA, 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 Rho A expressions in Tca8113 and SCC-4 cells were reduced significantly after transfection of Rho A-si RNA. Protein levels of galectin-3 and MMP-9 were also down-regulated significantly. Invasion ability was inhibited as well. Conclusion Rho Asi RNA can effectively inhibit Rho A 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 Rho A-si RNA can inhibit invasion of tongue carcinoma. Results show that Rho A may play an important role in the processes of invasion and metastasis of tongue carcinoma.

关 键 词：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 

分 类 号：R739.86[医药卫生—肿瘤]