Objective: 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.
Methods: 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.
Results: The integrated analysis of sequencing data and bioinformatics revealed that LIMCH1 exhibited significantly reduced mRNA (P < 0.001) and protein levels (P < 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 (P < 0.01), enhanced proliferation capacity over 72 hours relative to the control group (P < 0.01), and an increased apoptosis rate within 24 hours compared with the high expression group (P < 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 (P=0.013) and a higher lymph node metastasis rate (P < 0.05). Investigations into the potential mechanisms of action indicated that LIMCH1 did not influence the onset or progression of oral squamous cell carcinoma via the epithelial-mesenchymal transition pathway.
Conclusion: 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.
目的: 探究含LIM调宁蛋白同源域蛋白1(LIM and calponin homology domains 1，LIMCH1)在口腔鳞状细胞癌发生发展中的作用及潜在应用价值。
方法: 利用口腔鳞状细胞癌患者组织样本全转录组测序结合GO(Gene Ontology)、基因共表达网络等生物信息学分析方法筛选可能在口腔鳞状细胞癌发生发展中起关键作用的基因，使用12组口腔鳞状细胞癌患者组织样本采用实时定量PCR及Western blotting验证其表达趋势，采用CCK-8、流式细胞术、划痕实验等探究关键基因对口腔鳞状细胞癌细胞系Cal27及口腔潜在恶性病变细胞系DOK生物学行为的影响, 并采用TCGA(The Cancer Genome Atlas)中214例口腔鳞状细胞癌患者生存数据分析关键基因与患者临床疾病信息的关联, 同时结合基因集富集分析(gene set enrichment analysis，GSEA)的方法分析结果辅以实时定量PCR及Western blot探究关键基因可能的作用机制。
结果: 测序数据结合生物信息学分析发现LIMCH1在口腔鳞状细胞癌组织中mRNA(P＜0.001)及蛋白质(P＜0.01)表达显著低于正常对照组织。Cal27中LIMCH1低表达组24 h内划痕愈合面积大于高表达组(P＜0.01)，72 h内增殖能力高于高表达组(P＜0.01)，24 h凋亡率高于其高表达组(P＜0.05)，而DOK中LIMCH1低表达组与其高表达组差异无统计学意义。同时发现LIMCH1低表达与患者较差的远期预后(P=0.013)及较高的淋巴结转移率(P＜0.05)相关。对于LIMCH1潜在作用机制的探究中明确其并非通过上皮-间质转化途径影响口腔鳞状细胞癌的发生和发展。
结论: LIMCH1可能作为辅助口腔鳞状细胞癌预后判断的潜在生物学标志物，但其具体作用机制仍有待深入探究。
Keywords: LIM and calponin homology domains 1; Oral potentially malignant disorders; Oral squamous cell carcinoma.