West China Journal of Stomatology ›› 2021, Vol. 39 ›› Issue (3): 328-335.doi: 10.7518/hxkq.2021.03.013

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Effects of isoprenylcysteine carboxylmethyltransferase silencing on the migration and invasion of tongue squamous cell carcinoma

Zhou Nan1(), Chi Zengpeng1, Li Wenjian2, Zhao Kai3, Wang Shaoru2, Wang Qimin4, Tong Lei4, He Zongxuan5, Han Hongyu4, Wang Ying6, Chen Zhenggang4()   

  1. 1.College of Stomatology, Weifang Medical University, Weifang 261021, China
    2.School of Stomatology, Dalian Medical University, Dalian 116044, China
    3.School of Stomatology, Qingdao University, Qingdao 266003, China
    4.Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China
    5.Dept. of Oral and Maxillafacial Surgery, The Affiliated Hospital of Qingdao University, Qingdao 266005, China
    6.Dept. of Stomatology, Fourth People's Hospital of Jinan, Jinan 250031, China
  • Received:2020-06-18 Revised:2021-02-05 Online:2021-06-01 Published:2021-05-26
  • Contact: Chen Zhenggang E-mail:1123862614@qq.com;chenzhg1973@163.com
  • Supported by:
    The National Natural Science Foundation of China(81372908)

Abstract: Objective

The effect of isoprenylcysteine carboxymethyltransferase (ICMT) silencing on the migration and invasion of tongue squamous cell carcinoma was investigated by constructing the small interfering RNA (siRNA) of ICMT.

Methods

Through liposomal transfection, siRNA was transfected into human tongue squamous cell carcinoma CAL-27 and SCC-4 cells (ICMT-siRNA group) with a negative control group (transfected with NC-siRNA) and a blank control group (transfected with a transfection reagent but not with siRNA). Quantitative real-time polymerase chain reaction was performed to analyze the mRNA expression of ICMT and RhoA in each group of cells after transfection and to measure the silencing efficiency. Western blot was applied to examine the expression levels of ICMT, total RhoA, membrane RhoA, ROCK1, matrix metalloproteinase (MMP)-2, and MMP-9 proteins in each group. The migration and invasion abilities were evaluated via wound healing and Transwell motility assays.

Results

After CAL-27 and SCC-4 cells were transfected with ICMT-siRNA, the expression levels of ICMT genes and proteins decreased significantly in the experimental group compared with those in the negative and blank control groups (P<0.05). The mRNA and total protein expression levels of RhoA in the two groups were not significantly different (P>0.05). The expression levels of RhoA membrane protein, ROCK1, MMP-2, and MMP-9 decreased (P<0.05). The migration and invasion abilities were inhibited (P<0.05).

Conclusion

The migration and invasion abilities of CAL-27 and SCC-4 cells were reduced significantly after the transfection of ICMT-siRNA, and the involved mechanism might be related to the RhoA-ROCK signaling pathway.

Key words: isoprenylcysteine carboxylmethyltransferase, RhoA, tongue squamous cell carcinoma, migration, invasion

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