miR-let-7a-5p靶向调控SNAP23影响食管鳞状细胞癌发展的研究

doi:10.3969/j.issn.1004-616x.2025.01.011

癌变·畸变·突变 ›› 2025, Vol. 37 ›› Issue (1): 63-71.doi: 10.3969/j.issn.1004-616x.2025.01.011

• 肿瘤防治 • 上一篇

miR-let-7a-5p靶向调控SNAP23影响食管鳞状细胞癌发展的研究

陈娇¹, 刘清¹, 黄丛改², 郑树涛¹, 刘涛³, 卢晓梅¹

1. 新疆医科大学第一附属医院临床医学研究院, 省部共建中亚高发病成因与防治国家重点实验室, 新疆乌鲁木齐 830011;
2. 西南医科大学附属医院病理科, 四川泸州 646000;
3. 新疆医科大学第一附属医院检验科, 新疆乌鲁木齐 830011

收稿日期:2024-11-08 修回日期:2024-12-20 发布日期:2025-01-25
通讯作者: 卢晓梅
作者简介:陈娇，E-mail：2756761165@qq.com。
基金资助:
国家自然科学基金(82260568)；新疆维吾尔自治区“天山英才”培养计划科技创新领军人才项目(2022TSYCLJ0031)；新疆维吾尔自治区自然科学基金重点项目(2022D01D69)；新疆医科大学研究生创新创业项目(CXCY2024007)

miR-let-7a-5p targeted regulation of SNAP23 and malignant progression of esophageal squamous cell carcinoma

CHEN Jiao¹, LIU Qing¹, HUANG Conggai², ZHENG Shutao¹, LIU Tao³, LU Xiaomei¹

1. Clinical Medical Research Institute, The First Affiliated Hospital of Xinjiang Medical University, State Key Laboratory of Pathogenesis, Prevention and Treatment of High Incidence Diseases in Central Asia, Urumqi 830011, Xinjiang;
2. Department of Pathology, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, Sichuan;
3. Laboratory Department, The First Affiliated Hospital of Xinjiang Medical University, Urumqi 830011, Xinjiang, China

Received:2024-11-08 Revised:2024-12-20 Published:2025-01-25

摘要/Abstract

摘要： 目的：探讨微小RNA miR-let-7a-5p靶向调控突触体相关蛋白23(SNAP23)影响食管鳞状细胞癌(ESCC)发展的机制。方法：在人类癌症差异表达的miRNA数据库(dbDEMC)中分析了食管癌组织和癌旁正常组织中miR-let-7a-5p的表达水平，在人类癌症miRNA的交互式分析和可视化数据库(CancerMIRNome)中，采用Kaplan-Meier生存曲线及对数秩检验探讨了食管癌组织中miR-let-7a-5p的表达水平与患者预后之间的关系。采用实时荧光定量聚合酶链反应(RT-qPCR)法检测人永生化食管上皮细胞SHEE和ESCC细胞系中miR-let-7a-5p的表达；利用慢病毒载体转染技术，构建miR-let-7a-5p过表达的稳定转染细胞株；分别采用EdU细胞增殖实验、集落形成实验检测细胞增殖情况；细胞划痕实验检测细胞迁移；Transwell实验检测细胞侵袭。在dbDEMC和CancerMIRNome数据库中分析miR-let-7a-5p在食管癌患者外周血中的表达情况。在ENCORI、Diana Tarbase、miRDB数据库寻找miR-let-7a-5p的下游靶基因。Western blot法检测SNAP23蛋白的表达水平；双荧光素酶报告基因实验分析miR-let-7a-5p和SNAP23的靶向关系。结果：在食管癌组织中，miR-let-7a-5p的表达水平较癌旁正常组织降低，其表达水平与患者总生存期呈正相关(HR=0.57，P<0.05)。与人永生化食管上皮细胞SHEE相比，ESCC细胞系中miR-let-7a-5p呈低表达(P<0.01)。与对照组相比，增强miR-let-7a-5p表达后，ESCC细胞系TE-13的增殖、集落形成、迁移和侵袭能力均受到抑制(P<0.05或P<0.01)。在ESCC患者的外周血中，miR-let-7a-5p的表达水平较健康对照组增加(P<0.01)。分泌蛋白SNAP23被预测出是miR-let-7a-5p的下游靶基因。过表达miR-let-7a-5p后SNAP23的蛋白表达水平下降(P<0.01)。双荧光素酶报告基因实验结果显示，miR-let-7a-5p可下调野生型-SNAP23的相对荧光素酶活性(P<0.01)。结论： miR-let-7a-5p通过靶向下调SNAP23的表达，抑制ESCC的恶性生物学行为。

关键词: 食管鳞状细胞癌, miR-let-7a-5p, 突触体相关蛋白23, 侵袭转移, 靶向治疗

Abstract: OBJECTIVE： To explore the effect of miR-let-7a-5p on malignant development of esophageal squamous cell carcinoma (ESCC) by targeting and regulating the synaptosomal-associated protein of 23 kDa (SNAP23). METHODS： From database of Differentially Expressed MiRNAs in human Cancers (dbDEMC)， the expression levels of miR-let-7a-5p in cancer tissues and normal tissues adjacent to tumor were analyzed. From the database of MiRNAs in human Cancers (CancerMIRNome)，Kaplan-Meier survival curve and logarithmic rank test were used to investigate the relationship between the expression level of miR-let-7a-5p and the prognosis of patients with esophagea l cancer. Quantitative real-time reverse transcription-PCR (RT-qPCR) was used to detect the expression of miR-let-7a-5p in the human immortalized esophageal epithelial cell lines，SHEE and ESCC. Lentiviral vector transfection technology was utilized to construct stably transfected cell lines overexpressing miR-let-7a-5p. Cell proliferation was detected by the EdU cell proliferation and clone formation assay. Cell migration was detected by the cell scratch assay. The transwell assay was used to detect cell invasion. The expression of miR-let-7a-5p in peripheral blood of patients with esophageal cancer was analyzed in dbDEMC and CancerMIRNome databases. The downstream target genes of miR-let-7a-5p were searched in ENCORI，Diana Tarbase and miRDB databases. Protein expression of SNAP23 was detected by protein blotting (Western blot)；dual luciferase reporter gene assay was used to analyze the targeting relationship between miR-let-7a-5p and SNAP23. RESULTS： The expression level of miR-let-7a-5p in esophageal cancer tissues was lower than that in adjacent normal tissues，and the expression level was positively correlated with the overall survival of patients (P<0.05). The expression of miR-let-7a-5p in ESCC cell line was lower than that in human immortal SHEE (P<0.01). The proliferation，clone formation，migration and invasion abilities of TE-13 were significantly inhibited by enhanced miR-let-7a-5p expression compared with control (P<0.05 or P<0.01). In peripheral blood of ESCC patients，the expression level of miR-let-7a-5p was increased compared with that of healthy controls. The secreted protein SNAP23 was predicted to be the downstream target gene of miR-let-7a-5p. The protein expression level of SNAP23 decreased after overexpression of miR-let-7a-5p (P<0.01). The results of dual luciferase reporter gene assay showed that miR-let-7a-5p could down-regulate the relative luciferase activity of wild-type SNAP23 (P<0.01). CONCLUSION： miR-let-7a-5p can inhibit the malignant biological behavior of ESCC by down-regulating the expression of SNAP23.

Key words: esophageal squamous cell carcinoma, miR-let-7a-5p, SNAP23, invasive metastasis, targeted therapy

中图分类号:

R735.1

陈娇, 刘清, 黄丛改, 郑树涛, 刘涛, 卢晓梅. miR-let-7a-5p靶向调控SNAP23影响食管鳞状细胞癌发展的研究[J]. 癌变·畸变·突变, 2025, 37(1): 63-71.

CHEN Jiao, LIU Qing, HUANG Conggai, ZHENG Shutao, LIU Tao, LU Xiaomei. miR-let-7a-5p targeted regulation of SNAP23 and malignant progression of esophageal squamous cell carcinoma[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2025, 37(1): 63-71.

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miR-let-7a-5p靶向调控SNAP23影响食管鳞状细胞癌发展的研究

miR-let-7a-5p targeted regulation of SNAP23 and malignant progression of esophageal squamous cell carcinoma

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