Xu Li, Shi Wen, Li Yuehua, Shen Yajun, Xie Shang, Shan Xiaofeng, Cai Zhigang
Department of Oral and Maxillofacial Surgery, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing 100081, China.
First Clinical Division, Peking University School and Hospital of Stomatology & National Center for Stomatology & National Clinical Research Center for Oral Diseases & National Engineering Research Center of Oral Biomaterials and Digital Medical Devices, Beijing 100081, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2025 Feb 18;57(1):19-25. doi: 10.19723/j.issn.1671-167X.2025.01.004.
To explore the function of LIM and calponin homology domains 1 (LIMCH1) in the development and progression of oral squamous cell carcinoma (OSCC), along with their potential clinical applications.
By utilizing transcriptome sequencing data from two groups of oral squamous cell carcinoma patients, along with bioinformatics analytical techniques such as Gene Ontology (GO) and gene co-expression networks, we identified genes that might play a pivotal role in the pathogenesis of oral squamous cell carcinoma. We employed real-time quantitative PCR and Western blotting to validate the expression patterns of these genes across twelve patient tissue samples. Furthermore, we conducted CCK-8 assays, flow cytometry analyses, and scratch wound healing assays to assess the impact of key genes on the biological behaviors of both the Cal27 oral squamous cell carcinoma cell line and the potentially malignant DOK oral lesion cell line. Additionally, we examined correlations between these key genes and clinical disease parameters in 214 oral squamous cell carcinoma patients using The Cancer Genome Atlas (TCGA) data; gene set enrichment analysis (GSEA) analysis results were also incorporated to enhance our findings from real-time quantitative PCR and Western blotting regarding potential mechanisms underlying the action of these key genes.
The integrated analysis of sequencing data and bioinformatics revealed that exhibited significantly reduced mRNA ( < 0.001) and protein levels ( < 0.01) in the oral squamous cell carcinoma tissues compared with normal control tissues. In the Cal27 cells, the low LIMCH1 level group demonstrated a larger wound healing area within 24 hours than the control group ( < 0.01), enhanced proliferation capacity over 72 hours relative to the control group ( < 0.01), and an increased apoptosis rate within 24 hours compared with the high expression group ( < 0.05). However, no significant differences were observed between the low and high level groups in DOK cells. Furthermore, it was determined that low LIMCH1 level correlated with poor prognosis in the patients (=0.013) and a higher lymph node metastasis rate ( < 0.05). Investigations into the potential mechanisms of action indicated that did not influence the onset or progression of oral squamous cell carcinoma via the epithelial-mesenchymal transition pathway.
LIMCH1 level may function as a promising biomarker to aid in the prognostic assessment of oral squamous cell carcinoma; however, its precise mechanistic role requires further investigation.
探讨LIM和钙调蛋白同源结构域1(LIMCH1)在口腔鳞状细胞癌(OSCC)发生发展中的作用及其潜在的临床应用价值。
利用两组口腔鳞状细胞癌患者的转录组测序数据,结合基因本体论(GO)和基因共表达网络等生物信息学分析技术,鉴定可能在口腔鳞状细胞癌发病机制中起关键作用的基因。我们采用实时定量PCR和蛋白质免疫印迹法验证这些基因在12例患者组织样本中的表达模式。此外,我们进行了CCK-8实验、流式细胞术分析和划痕伤口愈合实验,以评估关键基因对Cal27口腔鳞状细胞癌细胞系和潜在恶性DOK口腔病变细胞系生物学行为的影响。另外,我们利用癌症基因组图谱(TCGA)数据研究了214例口腔鳞状细胞癌患者中这些关键基因与临床疾病参数之间的相关性;基因集富集分析(GSEA)分析结果也被纳入,以加强我们从实时定量PCR和蛋白质免疫印迹法中获得的关于这些关键基因作用潜在机制的发现。
测序数据与生物信息学的综合分析显示,与正常对照组织相比,口腔鳞状细胞癌组织中LIMCH1的mRNA水平(<0.001)和蛋白质水平(<0.01)显著降低。在Cal27细胞中,低LIMCH1水平组在24小时内的伤口愈合面积大于对照组(<0.01),在72小时内相对于对照组的增殖能力增强(<0.01),且与高表达组相比,24小时内的凋亡率增加(<0.05)。然而,在DOK细胞的低水平组和高水平组之间未观察到显著差异。此外,研究确定低LIMCH1水平与患者预后不良(=0.013)和较高的淋巴结转移率相关(<0.05)。对潜在作用机制的研究表明,LIMCH1不会通过上皮-间质转化途径影响口腔鳞状细胞癌的发生或发展。
LIMCH1水平可能作为一种有前景的生物标志物,有助于口腔鳞状细胞癌的预后评估;然而,其确切的作用机制仍需进一步研究。