West China Journal of Stomatology ›› 2021, Vol. 39 ›› Issue (1): 64-73.doi: 10.7518/hxkq.2021.01.010

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Effects of isoprenylcysteine carboxyl methyltransferase silencing on the proliferation and apoptosis of tongue squamous cell carcinoma

Wang Shaoru1,2(), Sun Wei1, Zhou Nan3, Zhao Kai4, Li Wenjian2, Chi Zengpeng3, Wang Ying5, Wang Qimin1, Tong Lei1, He Zongxuan6, Han Hongyu1, Chen Zhenggang1()   

  1. 1.Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China
    2.School of Stomatology, Dalian Medical University, Dalian 116044, China
    3.College of Stomatology, Weifang Medical University, Weifang 261021, China
    4.School of Stomatology, Qingdao University, Qingdao 266003, China
    5.Dept. of Stomatology, Fourth People, s Hospital of Jinan, Jinan 250031, China
    6.Dept. of Oral and Maxillafacial Surgery, The Affiliated Hospital of Qingdao University, Qingdao 266005, China
  • Received:2020-03-18 Revised:2020-10-27 Online:2021-02-01 Published:2021-03-02
  • Contact: Chen Zhenggang E-mail:wangxiaorukw@163.com;chenzhg1973@163.com
  • Supported by:
    The National Natural Science Foundation of China(81372908)

Abstract: Objective

This study aimed to explore the effects of silencing isoprenylcysteine carboxyl methyltransfe-rase (Icmt) through small interfering RNA (siRNA) interference on the proliferation and apoptosis of tongue squamous cell carcinoma (TSCC).


Three siRNA were designed and constructed for the Icmt gene sequence and were then transfected into TSCC cells CAL-27 and SCC-4 to silence Icmt expression. The tested cells were divided as follows: RNA interference groups Icmt-siRNA-1, Icmt-siRNA-2, and Icmt-siRNA-3, negative control group, and blank control group. The transfection efficiency of siRNA was detected by the fluorescent group Cy3-labeled siRNA, and the expression of Icmt mRNA was screened by quantitive real-time polymerase chain reaction (qRT-PCR) selected the experimental group for subsequent experiments. The expression of Icmt, RhoA, Cyclin D1, p21, extracellular regulated protein kinases (ERK), and phospho-extracellular regulated protein kinases (p-ERK) were analyzed by Western blot. The proliferation abilities of TSCC cells were determined by cell counting kit-8 assay. The change in apoptosis was detected by AnnexinV-APC/propidium staining (PI) assay. Cell-cycle analysis was conducted by flow cytometry.


The expression of Icmt mRNA and protein in TSCC cells significantly decreased after Icmt-siRNA transfection (P<0.05). No significant difference in RhoA mRNA and protein expression was detected (P>0.05), but the expression of RhoA membrane protein decreased compared with the negative control group and blank control groups (P<0.05). Cyclin D1 expression decreased, whereas p21 expression significantly increased and the relative expression of ERK protein in the experimental group did not significantly different that in the control group (P>0.05). However, the phosphorylation level of ERK was significantly reduced (P<0.05). The cell cycles of TSCC CAL-27 and SCC-4 were altered in G1/S, cell proliferation activity was inhibited, and apoptosis was induced (P<0.05).


Silencing Icmt can effectively downregulate its expression in TSCC cells, reduce the RhoA membrane targeting localization and cell proliferation, and induce apoptosis. Thus, Icmt may be a potential gene therapy target for TSCC.

Key words: isoprenylcysteine carboxyl methyltransferase, RhoA, tongue squamous cell carcinoma, cell proli-feration, cell cycle, apoptosis

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