West China Journal of Stomatology

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Effects of RhoA gene silencing by RNA interference on invasion of tongue carcinoma

Chen Zhenggang1,2, Tang Yongping1, Tong Lei1, Wang Ying3, Zhou Yuan3, Wang Qimin1, Han Jinhong1, He Zongxuan1, Liao Yixiang1, Fan Bing4, Zou Ronghai4, Zhang Jian4, Sun Xiaofeng4, Yan Guoxin4.   

  1. 1. Center of Stomatology, Qingdao Municipal Hospital Affiliated to Qingdao University Medical College, Qingdao 266071, China; 2. Dept. of Oral and Maxillofacial Surgery, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China; 3. College of Stomatology, Weifang Medical University, Weifang 261021, China; 4. Dept. of Stomatology, Wuxi No 2. People’s Hospital, Wuxi 214002, China
  • Received:2015-09-16 Revised:2015-12-18 Online:2016-04-01 Published:2016-04-01
  • Contact: Yan Guoxin, E-mail: yanguo_xin2015@163.com.

Abstract:

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.

Key words: RhoA gene, galectin-3, matrix metalloproteinase-9, RNA interference, tongue carcinoma, invasion