miR-204 Targets SOX4 to Inhibit the Development of Cutaneous Squamous Cell Carcinoma
Objective:To explore the function and possible molecular mechanism of miR-204 and SOX4 in cutaneous squamous cell carcinoma(CSCC).Methods:A total of 70 patients with CSCC who were diagnosed and underwent surgical treatment in the derma-tology department of the Second Affiliated Hospital of Air Force Military Medical University from May 2019 to March 2022 were selected.CSCC tissues and corresponding paracancer tissues of the patients were collected,and the mRNA levels of miR-204 and SOX4 in the tissue samples were detected by qRT-PCR.The protein level of SOX4 in tissue samples were detected by Western blot and immunohisto-chemical staining.The CSCC cell line SCL-1 was divided into control group,miR-NC group,miR-204 group,miR-204+pcDNA group and miR-204+SOX4 group.miR-NC,miR-204 mimics,miR-204 mimics and pcDNA3.1 empty vector,miR-204 mimics and pcDNA3.1+SOX4 vector were transfected into miR-NC group,miR-204 group,miR-204+pcDNA group and miR-204+SOX4 group by Lipofec-tamine 2000TM transfection reagents,respectively.And control group cells were not transfected.The targeting relationship between miR-204 and SOX4 was analyzed by dual luciferase reporter gene experiment.SCL-1 cell proliferation was detected by CCK-8 assay,cell cloning was detected by clonogenesis assay,cell apoptosis was detected by flow cytometry assay,and cell migration and invasion were detected by Transwell assay.The protein expression levels of SOX4,E-cadherin,N-cadherin and Vimentin were detected by West-em blot.Results:The expression of miR-204 was low in CSCC,and the expression of SOX4 mRNA and protein was high.The online analysis results of TargetScan software showed that binding sites existed at 5'of miR-204 and 3'of SOX4.Compared with the control group or the NC group,the relative proliferation activity,number of clone formation,migration and invasion of SCL-1 cells in miR-204 group were decreased(P＜0.05),and the apoptosis rate was increased(P＜0.05).The protein levels of SOX4,N-cadherin and Vimentin in SCL-1 cells were decreased(P＜0.05),while the protein level of E-cadherin was increased(P＜0.05).Compared with miR-204 group or miR-204+pcDNA group,the relative proliferation activity,number of clone formation,migration and invasion of SCL-1 cells in miR-204+SOX4 group were increased(P＜0.05),and the apoptosis rate was decreased(P＜0.05).The protein levels of SOX4,N-cadherin and Vimentin in SCL-1 cells were increased(P＜0.05),while the protein level of E-cadherin was decreased(P＜0.05).Conclusion:miR-204 inhibits the proliferation,migration,invasion and epithelial mesenchymal transformation of CSCC cells by targeting SOX4,and promotes cell apoptosis,thus alleviating the development of CSCC.
Cutaneous squamous cell carcinoma(CSCC)miR-204SRY-box transcription factor-4(SOX4)Proliferation and apop-tosisMigration and invasionEpithelial mesenchymal transformation(EMT)
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