PMID- 33723939 OWN - NLM STAT- MEDLINE DCOM- 20210317 LR - 20210319 IS - 2618-0456 (Electronic) IS - 1000-1182 (Print) IS - 1000-1182 (Linking) VI - 39 IP - 1 DP - 2021 Feb 1 TI - Effects of isoprenylcysteine carboxyl methyltransferase silencing on the proliferation and apoptosis of tongue squamous cell carcinoma. PG - 64-73 LID - 1000-1182(2021)01-0064-10 [pii] LID - 10.7518/hxkq.2021.01.010 [doi] AB - OBJECTIVES: 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). METHODS: 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. RESULTS: 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). CONCLUSIONS: 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. FAU - Wang, Shao-Ru AU - Wang SR AD - Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China. AD - School of Stomatology, Dalian Medical University, Dalian 116044, China. FAU - Sun, Wei AU - Sun W AD - Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China. FAU - Zhou, Nan AU - Zhou N AD - College of Stomatology, Weifang Medical University, Weifang 261021, China. FAU - Zhao, Kai AU - Zhao K AD - School of Stomatology, Qingdao University, Qingdao 266003, China. FAU - Li, Wen-Jian AU - Li WJ AD - School of Stomatology, Dalian Medical University, Dalian 116044, China. FAU - Chi, Zeng-Peng AU - Chi ZP AD - College of Stomatology, Weifang Medical University, Weifang 261021, China. FAU - Wang, Ying AU - Wang Y AD - Dept. of Stomatology, Fourth People,s Hospital of Jinan, Jinan 250031, China. FAU - Wang, Qi-Min AU - Wang QM AD - Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China. FAU - Tong, Lei AU - Tong L AD - Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China. FAU - He, Zong-Xuan AU - He ZX AD - Dept. of Oral and Maxillafacial Surgery, The Affiliated Hospital of Qingdao University, Qingdao 266005, China. FAU - Han, Hong-Yu AU - Han HY AD - Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China. FAU - Chen, Zheng-Gang AU - Chen ZG AD - Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China. LA - eng LA - chi PT - Journal Article PL - China TA - Hua Xi Kou Qiang Yi Xue Za Zhi JT - Hua xi kou qiang yi xue za zhi = Huaxi kouqiang yixue zazhi = West China journal of stomatology JID - 9422648 RN - 0 (RNA, Small Interfering) RN - EC 2.1.1.- (Protein Methyltransferases) RN - EC 2.1.1.100 (protein-S-isoprenylcysteine O-methyltransferase) SB - IM MH - Apoptosis MH - *Carcinoma, Squamous Cell MH - Cell Line, Tumor MH - Cell Proliferation MH - Humans MH - Protein Methyltransferases MH - RNA, Small Interfering MH - Tongue MH - *Tongue Neoplasms PMC - PMC7905405 OAB - OBJECTIVE: This study aimed to explore the effects of silencing isoprenylcysteine carboxyl methyltransferase (Icmt) through small interfering RNA (siRNA) interference on the proliferation and apoptosis of tongue squamous cell carcinoma (TSCC). METHODS: 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. RESULTS: 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). CONCLUSION: 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. OABL- eng OTO - NOTNLM OT - RhoA OT - apoptosis OT - cell cycle OT - cell proli-feration OT - isoprenylcysteine carboxyl methyltransferase OT - tongue squamous cell carcinoma COIS- 利益冲突声明：作者声明本文无利益冲突。 EDAT- 2021/03/17 06:00 MHDA- 2021/03/18 06:00 PMCR- 2021/02/01 CRDT- 2021/03/16 07:09 PHST- 2021/03/16 07:09 [entrez] PHST- 2021/03/17 06:00 [pubmed] PHST- 2021/03/18 06:00 [medline] PHST- 2021/02/01 00:00 [pmc-release] AID - 1000-1182(2021)01-0064-10 [pii] AID - wcjs-39-01-064 [pii] AID - 10.7518/hxkq.2021.01.010 [doi] PST - ppublish SO - Hua Xi Kou Qiang Yi Xue Za Zhi. 2021 Feb 1;39(1):64-73. doi: 10.7518/hxkq.2021.01.010.