| 胡新红,曹天宇,蔺建苹,吕雅洁,周 芳.miR-204靶向SOX4抑制皮肤鳞状细胞癌的发展[J].,2024,(14):2626-2633 |
| miR-204靶向SOX4抑制皮肤鳞状细胞癌的发展 |
| miR-204 Targets SOX4 to Inhibit the Development of Cutaneous Squamous Cell Carcinoma |
| 投稿时间：2024-01-29 修订日期：2024-02-27 |
| DOI：10.13241/j.cnki.pmb.2024.14.005 |
| 中文关键词: 皮肤鳞状细胞癌 miR-204 SRY相关HMG盒转录因子4（SOX4） 增殖和凋亡 迁移和侵袭 上皮间质转化 |
| 英文关键词: Cutaneous squamous cell carcinoma (CSCC) miR-204 SRY-box transcription factor-4 (SOX4) Proliferation and apoptosis Migration and invasion Epithelial mesenchymal transformation (EMT) |
| 基金项目:国家自然科学基金青年基金项目（81703119） |
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| 中文摘要: |
| 摘要 目的：探究miR-204和SRY相关HMG盒转录因子4（SOX4）在皮肤鳞状细胞癌（CSCC）中的功能以及可能的分子作用机制。方法：选择2019年5月至2022年3月于空军军医大学第二附属医院皮肤科确诊并接受手术治疗的皮肤鳞状细胞癌患者共70例，分别收集患者的CSCC组织和相应癌旁组织，采用qRT-PCR检测组织标本中miR-204和SOX4 mRNA的水平，Western blot和免疫组织化学染色检测组织标本中SOX4水平。将CSCC细胞系SCL-1分为对照组、miR-NC组、miR-204组、miR-204+pcDNA组和miR-204+SOX4组，采用Lipofectamine 2000TM转染试剂分别将miR-NC、miR-204模拟物、miR-204模拟物和pcDNA3.1空载体、miR-204模拟物和pcDNA3.1+SOX4载体分别转染至miR-NC组、miR-204组、miR-204+pcDNA组和miR-204+SOX4组，对照组细胞不转染。双荧光素酶报告基因实验检测miR-204与SOX4靶向关系。采用CCK-8法检测SCL-1细胞增殖，克隆形成实验检测SCL-1细胞克隆形成，流式细胞术实验检测SCL-1细胞凋亡，Transwell实验检测检测SCL-1细胞迁移和侵袭，Western blot检测SOX4、E-cadherin、N-cadherin和Vimentin蛋白表达水平。结果：CSCC组织中miR-204低表达，SOX4的mRNA和蛋白均高表达。TargetScan软件在线分析结果显示miR-204的5'端与SOX4的3'端存在结合位点。与对照组或NC组比较，miR-204组SCL-1细胞相对增殖活力、克隆形成数量、迁移和侵袭数量均降低（P<0.05），细胞凋亡率升高（P<0.05）；SCL-1细胞中SOX4、N-cadherin和Vimentin蛋白水平均降低（P<0.05），E-cadherin蛋白水平升高（P<0.05）。与miR-204组或miR-204+pcDNA组比较，miR-204+SOX4组SCL-1细胞相对增殖活力、克隆形成数量、迁移和侵袭数量均升高（P<0.05），细胞凋亡率降低（P<0.05）；SCL-1细胞中SOX4、N-cadherin和Vimentin蛋白水平均升高（P<0.05），E-cadherin蛋白水平降低（P<0.05）。结论：miR-204通过靶向SOX4抑制CSCC细胞的增殖、迁移、侵袭和上皮间质转化过程，并促进细胞凋亡，进而缓解CSCC的发展。 |
| 英文摘要: |
| ABSTRACT 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 dermatology 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 immunohistochemical 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 Lipofectamine 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 Western 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. |
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