Department of Tumor Radiotherapy, First People's Hospital of Fuyang, 311400 Hangzhou, China.
Department of General Medicine, First People's Hospital of Fuyang, 311400 Hangzhou, China.
Comput Math Methods Med. 2021 Apr 15;2021:5542485. doi: 10.1155/2021/5542485. eCollection 2021.
Protease-activated receptor-2 (PAR2) also known as F2RL1 is a G protein-coupled receptor that intimately correlates with cancer occurrence. DNA methylation turns out a vital mechanism regulating gene expression, while PAR2 promoter methylation is proven to be involved in cancer development. Hence, this study attempted to clarify the molecular mechanism by which PAR2 mediates lung adenocarcinoma (LUAD) progression, via identifying the effect of PAR2 promoter methylation on LUAD cell progression.
Associations of PAR2 promoter methylation with PAR2 gene expression and prognosis of LUAD patients were analyzed via bioinformatics analysis. PAR2 promoter methylation and gene expression at the cellular level were measured using methylation-specific PCR, qRT-PCR, and Western blot assays. DNA methyltransferase inhibitor 5-AzadC was used to treat cells to assess PAR2 gene expression alteration. Cell biological behaviors upon PAR2 overexpression were characterized via MTT, wound healing assay, and Transwell assay.
Bioinformatics analysis revealed that PAR2 promoter methylation was negatively related to PAR2 gene expression, while PAR2 promoter hypermethylation and low gene expression indicated favorable LUAD prognosis. Besides, it turned out that PAR2 presented upregulated expression and hypomethylated promoter in LUAD cells. Moreover, PAR2 gene expression was elevated in cells treated with 5-AzadC, and the proliferative, migratory, and invasive capabilities of cells with 5-AzadC or high PAR2 gene expression were all enhanced.
In sum, PAR2 promoter hypomethylation potentiates LUAD cell progression, in turn affecting the prognosis of LUAD patients.
蛋白酶激活受体-2（PAR2）也被称为 F2RL1，是一种与癌症发生密切相关的 G 蛋白偶联受体。DNA 甲基化是一种重要的基因表达调控机制，而 PAR2 启动子甲基化已被证明与癌症的发生发展有关。因此，本研究试图通过鉴定 PAR2 启动子甲基化对肺腺癌（LUAD）细胞进展的影响，阐明 PAR2 介导 LUAD 进展的分子机制。
通过生物信息学分析，分析 PAR2 启动子甲基化与 PAR2 基因表达及 LUAD 患者预后的关系。采用甲基化特异性 PCR、qRT-PCR 和 Western blot 检测 PAR2 启动子甲基化和细胞水平的基因表达。用 DNA 甲基转移酶抑制剂 5-AzadC 处理细胞，评估 PAR2 基因表达的变化。通过 MTT、划痕愈合实验和 Transwell 实验，研究 PAR2 过表达对细胞生物学行为的影响。
生物信息学分析显示，PAR2 启动子甲基化与 PAR2 基因表达呈负相关，而 PAR2 启动子高甲基化和低基因表达预示 LUAD 预后良好。此外，在 LUAD 细胞中，PAR2 表达上调且启动子低甲基化。此外，用 5-AzadC 处理细胞后，PAR2 基因表达上调，用 5-AzadC 或高 PAR2 基因表达处理的细胞增殖、迁移和侵袭能力均增强。
总之，PAR2 启动子低甲基化增强 LUAD 细胞的进展，进而影响 LUAD 患者的预后。
