Chen Zhenggang, Tang Yong-ping, Tong Lei, Wang Ying, Zhou Yuan, Wang Qimin, Han Jinhong, He Zongxuan, Liao Yixiang, Fan Bing, Zou Rong-hai, Zhang Jian, Sun Xiaofeng, Yan Guoxin
Hua Xi Kou Qiang Yi Xue Za Zhi. 2016 Apr;34(2):183-8. doi: 10.7518/hxkq.2016.02.016.
To study the effects of RhoA down-regulation by RNA interference on the invasion of tongue carcinoma Tca8113 and SCC-4.
Determination of the human RhoA sequence as well as the design and constructionof a short specific small interfering RNAs (siRNA) were performed. The siRNA of RhoA gene was transfected into humantongue squamous cell carcinoma Tca8113 and SCC-4 cells line by Lipofectamine 2000. Quantitative real-time polymerasechain reaction was used to examine the mRNA expressionlevels of RhoA. Protein expressions of mRNA, galectin-3,and matrix metalloproteinase (MMP)-9 were evaluated byWestern blot. Transwell invasion assay was performed toassess the invasion ability of tongue carcinoma.
RhoA expressions in Tca8113 and SCC-4 cells were reducedsignificantly after transfection of RhoA-siRNA. Protein levels f galectin-3 and MVP-9 were also down-regulated significantly. Invasion ability was inhibited as well.
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.
研究RNA干扰下调RhoA对舌癌Tca8113和SCC-4侵袭能力的影响。
测定人RhoA序列，设计并构建特异性小干扰RNA（siRNA）。采用脂质体2000将RhoA基因的siRNA转染至人舌鳞状细胞癌Tca8113和SCC-4细胞系。运用实时定量聚合酶链反应检测RhoA的mRNA表达水平。通过蛋白质印迹法评估mRNA、半乳糖凝集素-3和基质金属蛋白酶（MMP）-9的蛋白表达。采用Transwell侵袭实验评估舌癌的侵袭能力。
转染RhoA-siRNA后，Tca8113和SCC-4细胞中RhoA的表达显著降低。半乳糖凝集素-3和MMP-9的蛋白水平也显著下调。侵袭能力也受到抑制。
RhoA-siRNA可有效抑制Tca8113和SCC-4细胞中RhoA的表达。随着半乳糖凝集素-3和MMP-9蛋白表达下调，舌癌细胞的侵袭能力降低，表明RhoA-siRNA可抑制舌癌的侵袭。结果显示RhoA可能在舌癌侵袭和转移过程中发挥重要作用。
