微小RNA-302a-3p靶向溶酶体相关膜蛋白5抑制口腔鳞状细胞癌侵袭转移的研究
miR-302a-3p targeting lysosomal-associated membrane protein 5 inhibits the invasion and metastasis of oral squamous cell carcinoma
作者：余丽, 周铁军, 吴晓, 蔺辛红, 张晓艳, 赖永先, 廖新月, 斯航, 冯云, 简洁, 冯燕
Author：Yu Li, Zhou Tiejun, Wu Xiao, Lin Xinhong, Zhang Xiaoyan, Lai Yongxian, Liao Xinyue, Si Hang, Feng Yun, Jian Jie, Feng Yan
收稿日期：2024-11-08 年卷（期）页码：2025,43(4):547-547-558
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：微小RNA-302a-3p,溶酶体相关膜蛋白5,口腔鳞状细胞癌,侵袭转移,
Key words：microRNA-302a-3p,lysosomal-associated membrane protein 5,oral squamous cell carcinoma,inva-sion and metastasis,
基金项目：国家自然科学基金项目(82403404);四川省科学技术厅科技计划项目(2024JDRC0040);四川省医学青年创新课题计划(Q23028);泸州市科技计划项目(2023JYJ002);泸州市科技计划项目(2023SYF139);德阳市口腔医院-西南医科大学科技战略合作(2024DYKQXNYD-03);2024大学生创新创业计划项目(202410632009);2024大学生创新创业计划项目(2024368)
中文摘要
目的 探讨溶酶体相关膜蛋白5（LAMP5）和miR-302a-3p在口腔鳞状细胞癌（OSCC）中的表达及其对OSCC侵袭转移的影响。 方法 癌症基因组图谱（TCGA）数据库分析OSCC中LAMP5的表达及其作为预后指标的灵敏度。蛋白质印迹、定量聚合酶链反应（qRT-PCR）和细胞免疫染色检测LAMP5在OSCC组织及细胞中的表达。细胞计数试剂盒-8（CCK-8）、细胞免疫染色、迁移和侵袭实验评估LAMP5对细胞增殖、迁移和侵袭的影响。微小RNA（miR）靶向预测网站预测调控LAMP5的miR并采用双荧光素酶报告实验验证。裸鼠荷瘤模型评估敲低LAMP5对OSCC生长的影响。 结果 LAMP5在OSCC组织和细胞中高表达且具有较高的诊断灵敏度。敲低LAMP5显著抑制OSCC细胞的增殖、迁移和侵袭；而过表达LAMP5则促进OSCC细胞的增殖、迁移和侵袭。LAMP5的表达受miR-302a-3p的调控。敲低LAMP5显著抑制荷瘤小鼠OSCC肿瘤的生长。 结论 LAMP5通过增加OSCC细胞的增殖、迁移和侵袭促进OSCC的恶性进展，其表达受到miR-302a-3p因子的负向调控。
英文摘要
ObjectiveThis study aimed to explore the expression of lysosomal-associated membrane protein 5 (LAMP5) and microRNA (miR)-302a-3p in oral squamous cell carcinoma (OSCC) and their functional mechanism on the invasion and metastasis of OSCC.MethodsThe expression of LAMP5 in OSCC and its sensitivity as a prognostic indicator were analyzed on the basis of The Cancer Genome Atlas database. Western blot, quantitative reverse transcription polymerase chain reaction, and cell immunocytochemistry were used to detect the expression of LAMP5 in OSCC tissues and cells. The effect of LAMP5 on the proliferation, migration, and invasion of OSCC cells was evaluated through cell counting kit-8, immunocytochemistry, migration, and invasion assays, respectively. The miRNA targeting prediction websites were used to predict the miR that regulates LAMP5 and verify the targeted regulatory effect of miR-302a-3p on LAMP5. The effect of LAMP5 knockdown on OSCC tumor growth was evaluated in a nude mouse tumorigenesis model.ResultsLAMP5 was highly expressed in OSCC tissues and cells. It showed high sensitivity in the early diagnosis of OSCC. LAMP5 knockdown significantly inhibited the proliferation, migration, and invasion of OSCC cells, whereas LAMP5 overexpression increased these cell activities. The expression of LAMP5 was regulated by miR-302a-3p.In vivo, LAMP5 knockdown significantly inhibited the growth of OSCC tumor.ConclusionLAMP5 promotes the malignant progression of OSCC by enhancing the proliferation, migration, and invasion of OSCC cells. The expression of LAMP5 is negatively regulated by miR-302a-3p.
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