Expression of Cdc42 in tongue squamous cell carcinoma and its influence on cell proliferation, migration and invasion

doi:10.19438/j.cjoms.2023.03.001

Abstract

Abstract: PURPOSE: The study aimed to observe the expression of cell division cycle protein 42(Cdc42) in tongue squamous cell carcinoma (TSCC) and its relationship with clinicopathological features and prognosis, and explore the effect of silencing Cdc42 gene in vitro on proliferation, migration, invasion and epithelial-mesenchymal transformation (EMT) of TSCC and its mechanism. METHODS: The expression of Cdc42 gene in TSCC and adjacent tissues was detected by immunohistochemistry, the correlation between the expression of Cdc42 and clinicopathological parameters of TSCC and its influence on prognosis were analyzed. Human TSCC CAL27 and SCC-4 cells were randomly divided into 3 groups, namely Cdc42-siRNA group, negative control group and blank control group. Three small interfering RNA(siRNA) targeting human Cdc42 gene sequence were designed and constructed, Cdc42-siRNA and NC-siRNA was transfected into Cdc42-siRNA group and negative control group by liposome-mediated transfection, while the blank control group only added transfection reagent. Expression of Cdc42 mRNA and protein was detected by qRT-PCR and Western blot, and the Cdc42-siRNA transfection group with the best silencing effect was used as the follow-up experimental group. The expression of EMT and MAPK JNK/p38 pathway-related proteins was examined by Western blot. The proliferation, migration and invasion ability of cells in vitro were tested by CCK-8, wound healing test and Transwell invasion test. SPSS 22.0 software package was used for data analysis. RESULTS: The positive expression rate of Cdc42 was 70.5%(31/44) in 44 patients with TSCC, and 38.6%(17/44) in adjacent tissues, the difference was statistically significant(P<0.05). The expression of Cdc42 was related to cervical lymph node metastasis, clinical stage and depth of invasion(P<0.05). Kaplan-Meier survival analysis showed that the prognosis of TSCC patients with positive expression of Cdc42 was not significantly different from those with negative expression(P>0.05). In vitro experiments, in Cdc42-siRNA group, the expression of Cdc42 mRNA and protein decreased significantly(P<0.05), the relative expression of EMT-related protein mesenchymal markers N-cadherin and Vimentin decreased significantly(P<0.05), and the relative expression of MMP-9 decreased significantly(P<0.05); however, the relative expression of E-cadherin was increased significantly(P<0.05). The relative expression levels of p38-MAPK and JNK proteins related to MAPK JNK/p38 pathway had no significant difference(P>0.05), but the expression levels of p-p38-MAPK and p-JNK proteins decreased significantly (P<0.05). In addition, the ability of proliferation, invasion and migration of cells in Cdc42-siRNA group decreased significantly(P<0.05). CONCLUSIONS: The expression of Cdc42 increased in TSCC patients, and the expression was related to cervical lymph node metastasis, clinical stage and depth of invasion. The silencing of Cdc42 may inhibit the transformation of EMT process of TSCC cells by blocking MAPK JNK/p38 pathway, thus inhibiting its invasion and migration, and negatively regulating TSCC cell proliferation.

Key words: Cdc42, Tongue squamous carcinoma cell, Proliferation, Invasion, Migration, Epithelial-mesenchymal transformation

CLC Number:

R739.8

MU Jia-ning, YUAN Wen-qing, LU Zhou, ZHAO Yang, WANG Qi-min, TONG Lei, CHEN Zheng-gang. Expression of Cdc42 in tongue squamous cell carcinoma and its influence on cell proliferation, migration and invasion[J]. China Journal of Oral and Maxillofacial Surgery, 2023, 21(3): 209-217.

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