Effects of silencing Icmt on the proliferation, apoptosis, and cell cycle of CAL-27 and SCC-4 cells

doi:10.19438/j.cjoms.2021.02.001

Abstract

Abstract: PURPOSE: This study was aimed to explore the effects and regulatory mechanisms of silencing Icmt on cell proliferation, apoptosis, and cell cycle of cell line CAL-27 and SCC-4 in vitro. METHODS: Three siRNAs were designed and constructed for Icmt gene sequence, and then transfected into CAL-27 and SCC-4 cells to silence Icmt expression. The tested cells were divided as follows: RNA interference groups including Icmt-siRNA-1, Icmt-siRNA-2, and Icmt-siRNA-3, negative control group, and blank control group. The mRNA and protein expression of Icmt and K-Ras were examined by real-time PCR and Western blot, respectively. The expression of Cyclin D1, p21, Akt, and p-Akt were ex-amined by Western blot. The proliferation abilities of CAL-27 and SCC-4 cells were determined by cell counting kit-8 assay. Cell cycle analysis and apoptosis abilities of CAL-27 and SCC-4 cells were detected by flow cytometry. Statistical analysis and presentation was performed using GraphPad Prism 8.2.1 software. RESULTS: The expression of Icmt mRNA and protein in CAL-27 and SCC-4 cells was reduced significantly after Icmt siRNAs were transfected (P<0.05). No significant difference in K-Ras mRNA and protein expression was detected(P>0.05), but the expression of K-Ras membrane protein was decreased significantly compared with the negative control group and the blank control group (P<0.05). Cyclin D1 expression was decreased, whereas p21 expression was increased significantly. The expression of Akt was invariant (P>0.05), but the expression of p-Akt was significantly decreased (P<0.05). The cell cycle was altered in G1/S, the growth-proliferative activity was inhibited and apoptosis was significantly induced (P<0.05). CONCLUSIONS: Silencing Icmt can effectively reduce the proliferation and induce apoptosis of CAL-27 and SCC-4 cells by affecting K-Ras membrane targeting localization, and then negatively regulating cell cycle and down-regulating K-Ras /PI3K/Akt/mTOR signaling pathway.

Key words: Isoprenylcysteine carboxyl methyltransferase, K-Ras, Tongue squamous cell carcinoma, CAL-27, SCC-4, Cell proliferation, Cell cycle, Cell apoptosis

CLC Number:

R739.8

CHEN Zheng-gang, WANG Qi-min, TONG Lei, WANG Yun-ying, XU Xiao-na, WANG Ying, HAN Hong-yu, SHENG Shan-gui, WANG Shao-ru. Effects of silencing Icmt on the proliferation, apoptosis, and cell cycle of CAL-27 and SCC-4 cells[J]. China Journal of Oral and Maxillofacial Surgery, 2021, 19(2): 97-104.

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