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

doi:10.7518/hxkq.2021.03.013

Abstract

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

CLC Number:

R 782

Zhou Nan, Chi Zengpeng, Li Wenjian, Zhao Kai, Wang Shaoru, Wang Qimin, Tong Lei, He Zongxuan, Han Hongyu, Wang Ying, Chen Zhenggang. Effects of isoprenylcysteine carboxylmethyltransferase silencing on the migration and invasion of tongue squamous cell carcinoma[J]. West China Journal of Stomatology, 2021, 39(3): 328-335.

Figures/Tables 6

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ICMT-siRNA	产品编号	产品名称	靶序列
ICMT-siRNA-1	StB0010313A	genOFFTMst-h-ICMT_001	GAAGAAGAAAT-CTCACTAA
ICMT-siRNA-2	StB0010313B	genOFFTMst-h-ICMT_002	GTTAGAGTTCACACTTGAA
ICMT-siRNA-3	StB0010313C	genOFFTMst-h-ICMT_003	CTTCCGCGATCGAACAGAA

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