Feng Jing, He Wei, Fang Jin
Department of Stomatology, The Affiliated Huaian No. 1 People's Hospital of Nanjing Medical University, Huaian, 223300, Jiangsu, China.
Department of Stomatology, The People's Hospital of Suichang Country, Lishui, 323300, Zhejiang, China.
Sci Rep. 2025 May 31;15(1):19179. doi: 10.1038/s41598-025-02172-y.
To delve into the role and molecular mechanism of lysosome-associated membrane protein 3 (LAMP3) in tongue squamous cell carcinoma (TSCC). Differential expression of LAMP3 in TSCC from GEO microarray was analyzed, and pathway enrichment analysis of LAMP3 was performed utilizing GSEA. LAMP3 expression was detected by western blot, immumohistochemical staining and qRT-PCR. LAMP3 knockdown plasmid was constructed for a variety of biological function assays to verify the involvement of LAMP3 in TSCC. The upstream transcription factors and binding sites of LAMP3 were bioinformatically predicted. Dual luciferase reporter gene assay was applied to check the presence of c-Myc binding to the promoter region of LAMP3 and the regulation of its transcription. Further, a xenograft tumor model was developed to corroborate the impact of LAMP3 on tumor growth in vivo. LAMP3 expression was enhanced in TSCC tissues and cells. LAMP3 knockdown attenuated TSCC cell proliferation, DNA replication and metastatic capacity while induced glucose metabolism reprogramming in vitro. Meanwhile, LAMP3 depletion contributed to the delay of tumor progression in vivo. c-Myc was found bind to the LAMP3 promoter region to positively modulate LAMP3 transcriptional expression. LAMP3 regulated by c-Myc enhanced TSCC cell proliferation, DNA replication capacity while induced glucose metabolism reprogramming, suggesting a potential target for clinical trials in TSCC.
探究溶酶体相关膜蛋白3(LAMP3)在舌鳞状细胞癌(TSCC)中的作用及分子机制。分析来自GEO微阵列的TSCC中LAMP3的差异表达,并利用GSEA对LAMP3进行通路富集分析。通过蛋白质免疫印迹、免疫组织化学染色和qRT-PCR检测LAMP3表达。构建LAMP3敲低质粒用于各种生物学功能检测,以验证LAMP3在TSCC中的作用。通过生物信息学预测LAMP3的上游转录因子和结合位点。应用双荧光素酶报告基因检测来检查c-Myc与LAMP3启动子区域的结合情况及其转录调控。此外,建立异种移植肿瘤模型以证实LAMP3对体内肿瘤生长的影响。LAMP3在TSCC组织和细胞中表达增强。LAMP3敲低减弱了TSCC细胞增殖、DNA复制和转移能力,同时在体外诱导葡萄糖代谢重编程。同时,LAMP3缺失导致体内肿瘤进展延迟。发现c-Myc与LAMP3启动子区域结合以正向调节LAMP3转录表达。由c-Myc调控的LAMP3增强了TSCC细胞增殖、DNA复制能力,同时诱导葡萄糖代谢重编程,提示其可能是TSCC临床试验的潜在靶点。