METHODS: The expression and consensus cluster analyses of m6A methylation regulators in early-stage LUAD were performed. The clinicopathological features, immune cell infiltration, survival and functional enrichment in different subtypes were analyzed. We also constructed a prognostic model. Clinical tissue samples were used to validate the expression of model genes through real-time polymerase chain reaction (RT-PCR). In addition, cell scratch assay and Transwell assay were also performed.
RESULTS: Expression of m6A methylation regulators was abnormal in early-stage LUAD. According to the consensus clustering of m6A methylation regulators, patients with early-stage LUAD were divided into two subtypes. Two subtypes showed different infiltration levels of immune cell and survival time. A prognostic model consisting of HNRNPC, IGF2BP1 and IGF2BP3 could be used to predict the survival of early-stage LUAD. RT-PCR results showed that HNRNPC, IGF2BP1 and IGF2BP3 were significantly up-regulated in early-stage LUAD tissues. The results of cell scratch assay and Transwell assay showed that overexpression of HNRNPC promotes the migration and invasion of NCI-H1299 cells, while knockdown HNRNPC inhibits the migration and invasion of NCI-H1299 cells.
CONCLUSIONS: This work reveals that m6A methylation regulators may be potential biomarkers for prognosis in patients with early-stage LUAD. Our prognostic model may be of great value in predicting the prognosis of early-stage LUAD.
方法:进行了早期LUAD中m6A甲基化调节因子的表达和一致性聚类分析。临床病理特征,免疫细胞浸润,分析了不同亚型的存活和功能富集。我们还构建了一个预后模子。临床组织样本用于通过实时聚合酶链反应(RT-PCR)验证模型基因的表达。此外,还进行了细胞划痕测定和Transwell测定。
结果:在早期LUAD中m6A甲基化调节因子的表达异常。根据m6A甲基化调节因子的共识聚类,早期LUAD患者分为两种亚型.两种亚型表现出不同的免疫细胞浸润水平和存活时间。由HNRNPC组成的预后模型,IGF2BP1和IGF2BP3可用于预测早期LUAD的生存。RT-PCR结果显示,HNRNPC,IGF2BP1和IGF2BP3在早期LUAD组织中显著上调。细胞划痕实验和Transwell实验结果表明,HNRNPC过表达促进NCI-H1299细胞的迁移和侵袭,而敲低HNRNPC抑制NCI-H1299细胞的迁移和侵袭。
结论:这项工作揭示了m6A甲基化调节因子可能是早期LUAD患者预后的潜在生物标志物。我们的预后模型可能对预测早期LUAD的预后具有重要价值。