Ma Jinhua, Hu Xiaodong, Dai Baoqiang, Wang Qiang, Wang Hongqin
Department of Otolaryngology, Cangzhou Central Hospital, Cangzhou, Hebei, China.
PeerJ. 2021 Apr 14;9:e11259. doi: 10.7717/peerj.11259. eCollection 2021.
Laryngeal squamous cell carcinoma (LSCC) is the second most aggressive head and neck squamous cell carcinoma. Although much work has been done to optimize its treatment, patients with LSCC still have poor prognosis. Therefore, figuring out differentially expressed genes (DEGs) contained in the progression of LSCC and employing them as potential therapeutic targets or biomarkers for LSCC is extremely meaningful.
Overlapping DEGs were screened from two standalone Gene Expression Omnibus datasets, and Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses were performed. By applying STRING and Cytoscape, a protein-protein network was built, and module analysis was carried out. The hub genes were selected by maximal clique centrality with the CytoHubba plugin of Cytoscape. UALCAN and GEPIA data were examined to validate the gene expression findings. Moreover, the connection of the hub genes with LSCC patient overall survival was studied employing The Cancer Genome Atlas. Then, western blot, qRT-PCR, CCK-8, wound healing and transwell assays were bring to use for further verify the key genes.
A total of 235 DEGs were recorded, including 83 upregulated and 152 downregulated genes. A total of nine hub genes that displayed a high degree of connectivity were selected. UALCAN and GEPIA databases verified that these genes were highly expressed in LSCC tissues. High expression of the SPP1, SERPINE1 and Matrix metalloproteinases 1 (MMP1) genes was connected to worse prognosis in patients with LSCC, according to the GEPIA online tool. Western blot and qRT-PCR testify SPP1, SERPINE1 and MMP1 were upregulated in LSCC cells. Inhibition of SPP1, SERPINE1 and MMP1 suppressed cell proliferation, invasion and migration.
The work here identified effective and reliable diagnostic and prognostic molecular biomarkers by unified bioinformatics analysis and experimental verification, indicating novel and necessary therapeutic targets for LSCC.
喉鳞状细胞癌（LSCC）是第二大侵袭性头颈部鳞状细胞癌。尽管在优化其治疗方面已经做了很多工作，但LSCC患者的预后仍然很差。因此，找出LSCC进展过程中包含的差异表达基因（DEGs）并将其用作LSCC的潜在治疗靶点或生物标志物具有极其重要的意义。
从两个独立的基因表达综合数据库中筛选重叠的DEGs，并进行基因本体论和京都基因与基因组百科全书通路富集分析。通过应用STRING和Cytoscape构建蛋白质-蛋白质网络，并进行模块分析。使用Cytoscape的CytoHubba插件通过最大团中心性选择枢纽基因。检查UALCAN和GEPIA数据以验证基因表达结果。此外，利用癌症基因组图谱研究枢纽基因与LSCC患者总生存期的关系。然后，使用蛋白质免疫印迹、qRT-PCR、CCK-8、伤口愈合和Transwell实验进一步验证关键基因。
共记录了235个DEGs，包括83个上调基因和152个下调基因。总共选择了9个具有高度连通性的枢纽基因。UALCAN和GEPIA数据库验证了这些基因在LSCC组织中高表达。根据GEPIA在线工具，SPP1、SERPINE1和基质金属蛋白酶1（MMP1）基因的高表达与LSCC患者的较差预后相关。蛋白质免疫印迹和qRT-PCR证明SPP1、SERPINE1和MMP1在LSCC细胞中上调。抑制SPP1、SERPINE1和MMP1可抑制细胞增殖、侵袭和迁移。
本研究通过统一的生物信息学分析和实验验证，确定了有效且可靠的诊断和预后分子生物标志物，为LSCC指明了新的必要治疗靶点。
