| Objective:Although the size of the larynx is small,it is responsible for the important physiological functions of human body,such as vocal production,breathing,swallowing and so on.Laryngeal squamous cell carcinoma(LSCC)ranks second in head and neck cancer and respiratory tract malignant tumors,and has a high incidence in northern China.At present,surgery is still the main treatment for the disease,and in situ recurrence and easy metastasis are the main risk factors for poor prognosis.In this study,the relationship between the expression of hsa-miR-1207-5p and clinicopathological parameters of patients with LSCC was revealed through cell lines in vitro,transplanted tumor models in nude mice and clinical samples,and the biological function and downstream regulatory mechanism of hsa-miR-1207-5p and its target gene spindle and centromere associated protein 3(SKA3)on the malignant phenotype of LSCC were discussed.Methods:The expression of hsa-miR-1207-5p in LSCC and adjacent normal mucosal(ANM)tissues were analyzed via in situ hybridization and quantitative PCR(q PCR),and clinical significance was analyzed by bioinformatics.The function of hsa-miR-1207-5p in LSCC was investigated by gene overexpression,proliferation analysis,monoclonal assay,scratch assay,Transwell assay and flow cytometry.The xenograft tumor model of LSCC in nude mice was established and hsa-miR-1207-5p agomir was injected into the tumor to evaluate the regulatory effect of hsa-miR-1207-5p on tumorigenesis of LSCC in vivo.Target genes of hsa-miR-1207-5p were predicted,and then functional annotation was performed.In addition,the regulatory effect of hsa-miR-1207-5p on SKA3,the predicted target gene,was verified by q PCR,western blot,luciferase and immunohistochemistry.The expression and clinical significance of SKA3 were analyzed in TCGA database and transcriptional data of 150 LSCC and ANM tissues.The phenotypic changes of LSCC cells were observed after knock-down SKA3,in FD-LSC-1 and TU-177 cells.Next,we designed a rescue experiment to inspect whether hsa-miR-1207-5p inhibits LSCC cell malignant behavior via downregulating the target gene SKA3.Results:Hsa-miR-1207-5p was downregulated in LSCC tissues and cells,which was correlated with the malignant progression and poor prognosis of LSCC.Overexpression of hsa-miR-1207-5p inhibited proliferation,migration,invasion and in vivo tumorigenesis of LSCC cells,affected the cell cycle progression.In vivo experiments indicated that hsa-miR-1207-5p could notably restrain the growth of LSCC cells.The results of GO and KEGG pathway enrichment analysis showed,hsa-miR-1207-5p target genes were involved in cell cycle regulation,proliferation,adhesion,PI3K/Akt pathway and other biological processes.Luciferase report showed that hsa-miR-1207-5p directly combined with SKA3 3’UTR and downregulated SKA3 expression.Combined with transcriptome sequencing data and TCGA database data analysis,it was found that the expression of SKA3 was significantly upregulated in LSCC,and the high expression of SKA3 was associated with malignant progression and poor prognosis of 32 kinds of tumors such as LSCC.The results of cell function study showed that SKA3 played a completely opposite function to hsa-miR-1207-5p in LSCC.Rescue experiments evidenced that hsa-miR-1207-5p inhibits suppresses the malignant phenotypes of LSCC through SKA3.In addition,overexpression of hsa-miR-1207-5p and knockdown of SKA3 inhibited the migration and invasion of LSCC by regulating epithelial–mesenchymal transition(EMT).Conclusion:The low expression of hsa-miR-1207-5p is related to the malignant progression of LSCC.Hsa-miR-1207-5p inhibits the proliferation,migration and invasion of LSCC cells by down-regulating the expression of SKA3.Hsa-miR-1207-5p/SKA3 axis may be play an important biological role in the malignant progression of LSCC and is expected to be a new marker and target for the diagnosis and therapy of LSCC. |
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