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

doi:10.7518/hxkq.2025.2024409

华西口腔医学杂志 ›› 2025, Vol. 43 ›› Issue (4): 547-558.doi: 10.7518/hxkq.2025.2024409

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

余丽¹^,²(), 周铁军³, 吴晓², 蔺辛红², 张晓艳¹^,², 赖永先²^,⁴, 廖新月²^,⁵, 斯航²^,⁵, 冯云²^,⁵, 简洁⁶(), 冯燕²^,⁵()

^1.西南医科大学附属口腔医院牙周黏膜病科，泸州 646000
^2.口颌面修复重建和再生泸州市重点实验室，泸州 646000
^3.西南医科大学附属医院病理科，泸州 646000
^4.西南医科大学附属口腔医院预防保健科，泸州 646000
^5.西南医科大学附属口腔医院儿童口腔科，泸州 646000
^6.德阳市口腔医院，德阳 618000

收稿日期:2024-11-08 修回日期:2025-01-23 出版日期:2025-08-01 发布日期:2025-08-29
通讯作者: 简洁,冯燕 E-mail:yulixt126@swmu.edu.cn;jianjie_4950@163.com;fyxt126@swmu.edu.cn
作者简介:余丽，主治医师，博士，E-mail：yulixt126@swmu.edu.cn
基金资助:
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)

miR-302a-3p targeting lysosomal-associated membrane protein 5 inhibits the invasion and metastasis of oral squamous cell carcinoma

Yu Li¹^,²(), Zhou Tiejun³, Wu Xiao², Lin Xinhong², Zhang Xiaoyan¹^,², Lai Yongxian²^,⁴, Liao Xinyue²^,⁵, Si Hang²^,⁵, Feng Yun²^,⁵, Jian Jie⁶(), Feng Yan²^,⁵()

^1.Dept. of Periodontal and Oral Mucosal Disease, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China
^2.Oral & Maxillofacial Reconstruction and Regeneration of Luzhou Key Laboratory, Luzhou 646000, China
^3.Dept. of Pathology, The Affiliated Hospital of Southwest Medical University, Luzhou 646000, China
^4.Dept. of Preventive Health Care, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China
^5.Dept. of Pediatric Dentistry, The Affiliated Stomatology Hospital of Southwest Medical University, Luzhou 646000, China
^6.Deyang Stomatological Hospital, Deyang 618000, China

Received:2024-11-08 Revised:2025-01-23 Online:2025-08-01 Published:2025-08-29
Contact: Jian Jie,Feng Yan E-mail:yulixt126@swmu.edu.cn;jianjie_4950@163.com;fyxt126@swmu.edu.cn
Supported by:
National Natural Science Foundation of China(82403404);Sichuan Science and Technology Program(2024JDRC0040);Youth Innovation Project of Sichuan Provincial Medical Association(Q23028);Luzhou Science and Technology Program(2023JYJ002);Deyang Stomatological Hospital-Southwest Medical University Science and Technology Strategic Cooperation Project(2024DYKQXNYD03);2024 National and School Undergraduate Innovation and Entrepreneurship Program(202410632009)

摘要/Abstract

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。

关键词: 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

Abstract:

Objective This 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. Methods The 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. Results LAMP5 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. Conclusion LAMP5 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.

Key words: microRNA-302a-3p, lysosomal-associated membrane protein 5, oral squamous cell carcinoma, inva-sion and metastasis

中图分类号:

R739.8

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. 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[J]. 华西口腔医学杂志, 2025, 43(4): 547-558.

Yu Li, Zhou Tiejun, Wu Xiao, Lin Xinhong, Zhang Xiaoyan, Lai Yongxian, Liao Xinyue, Si Hang, Feng Yun, Jian Jie, Feng Yan. miR-302a-3p targeting lysosomal-associated membrane protein 5 inhibits the invasion and metastasis of oral squamous cell carcinoma[J]. West China Journal of Stomatology, 2025, 43(4): 547-558.

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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

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