| Objective Oral squamous cell carcinoma(OSCC),as the most common malignant cancer in the oral cavity and maxillofacial region,has a very high recurrence rate.Although more and more studies have been conducted to gain an in-depth understanding of OSCC,the exact identifying molecules of OSCC are still unknown.Therefore,finding new molecular targets is very important to improve the therapeutic effect of OSCC.Previous research reports have shown that the protein voltage-gated sodium channels(VGSC)Nav1.5 are highly expressed in various types of cancer and are involved in promoting the proliferation,migration and invasion of cancer cells.However,the regulatory mechanism of Nav1.5 in cancer remains unclear.In this study,we found that Nav1.5 is highly expressed in both tissues and cells of OSCC.By analyzing the clinical characteristics of the patients,we found that the expression level of Nav1.5 and the neutrophil to lymphocyte ratio(NLR),platelet to lymphocyte ratio(PLR),and stage of tumor lymph node metastasis(TNM)Etc.are closely related.In addition,we found that Nav1.5 is mainly localized on the cell membrane,and knocking down the expression level of Nav1.5 can promote the apoptosis of HSC-3(human oral squamous cell carcinoma)cells and reduce the proliferation of HSC-3 cells.The results of Transwell experiments showed that knocking down the expression level of Nav1.5can effectively inhibit the migration and invasion of HSC-3 cells.Furthermore,we found that si-Nav1.5 inhibited the protein and m RNA expression levels of the key?-catenin genes in the Wnt /?-catenin signaling pathway,cyclin D1 and c-Myc downstream target genes.In summary,these results indicate that Nav1.5 may participate in the process of OSCC through the Wnt /?-catenin signaling pathway.Methods Eight primary OSCC patients who had not undergone chemoradiotherapy in the Oral and Maxillofacial Surgery of the First Affiliated Hospital of Anhui Medical University were selected.Part of the tumor tissue removed during the operation was collected as the experimental group,and the adjacent part of the tumor tissue was normal Tissue was used as a control group.The study was implemented with the approval of the Ethics Committee of Anhui Medical University.By immunohistochemical staining and Western blotting analysis,at the OSCC tissue level,the expression levels of Nav1.5 in the experimental group and the control group were different.In addition,the clinical characteristics of 8 patients with OSCC and the correlation between Nav1.5 expression levels were analyzed.Secondly,by analyzing the differences in Nav1.5 expression levels in four different OSCC cell lines,a cell line(HSC-3)with the highest Nav1.5 expression level was selected for subsequent series of cytological studies.In addition,the expression localization of Nav1.5 in HSC-3 cells after immunofluorescence staining was observed with a laser copolymer gel microscope.Real-time polymerase chain reaction(RT-PCR),Western blotting analysis and immunofluorescence staining were used to determine the silencing efficiency of siRNA(small interference RNA)on Nav1.5 expression level.The effects of si-Nav1.5 on the biological behavior(including cell proliferation,apoptosis,cell cycle,metastasis,and invasion)of HSC-3 cells were measured by CCK-8,flow cytometry,and Transwell experiments.QRT-PCR and Western blotting analysis were used to detect m RNA and protein expression levels of Wnt /?-catenin signaling pathway-related proteins(including key pathway genes ?-catenin and downstream target genes cyclin D1,c-Myc)after transfection..SPSS 19.0 software was used to analyze the experimental data,and the measured data was expressed as "x ± SD".All data were statistically analyzed using t test or one-way analysis of variance.P <0.05 indicates that the difference in data is statistically significant.Results Immunohistochemistry and Western blotting analysis showed that Nav1.5 was highly expressed in oral squamous cell carcinoma tissues,and lowly expressed in adjacent normal tissues.In addition,the expression level of Nav1.5 is higher in the neutrophil-to-lymphocyte ratio(NLR)and the platelet-to-lymphocyte ratio(PLR)),Higher tumor lymph node metastasis stage(TNM stage,Tumor-Node-Metastasis stage)and higher in tissues with lymph node metastasis.In addition,the results of immunofluorescence of Nav1.5 in HSC-3 cells revealed that Nav1.5 was mainly localized on the cytoplasm and cell membrane.In addition,si-Nav1.5 can significantly reduce the proliferation ability of HSC-3 cells and promote their apoptosis,as well as induce the accumulation of G1 phase cells in the cell cycle.The results of Transwell experiments showed that si-Nav1.5 can significantly reduce the ability of HSC-3 cells to metastasize and invade.In addition,si-Nav1.5 inhibited the expression of key genes?-catenin in the Wnt /?-catenin signaling pathway and downstream target genes cyclin D1 and c-Myc of the pathway.Conclusions Si-Nav1.5 inhibits the biological behavior of oral squamous cell carcinoma through the Wnt /?-catenin signaling pathway... |
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