新型基质/免疫评分相关 P2RY12 在肺腺癌微环境中的预后价值及免疫浸润分析。

Prognostic value and immune infiltration of a novel stromal/immune score-related P2RY12 in lung adenocarcinoma microenvironment.

机构信息

Department of Pulmonary Medicine, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai, China.

出版信息

Int Immunopharmacol. 2021 Sep;98:107734. doi: 10.1016/j.intimp.2021.107734. Epub 2021 Jun 25.

DOI:10.1016/j.intimp.2021.107734

PMID:34175738

Abstract

OBJECTIVE

Increasing evidence highlights the clinical implications of P2RY12 that belongs to the family of G-protein coupled receptors in carcinogenesis. Here, this study was designed to explore the associations between P2RY12 and tumor immune microenvironment of lung adenocarcinoma (LUAD).

METHODS

Based on 352 LUAD samples from The Cancer Genome Atlas (TCGA), stromal and immune scores of each sample were estimated by ESTIMATE algorithm. Differential expression analysis was presented between stromal/immune high- and low-score groups. Protein-protein interaction (PPI) was then constructed by STRING database. Univariate and multivariate Cox regression analysis was utilized to screen prognosis-related factors. Co-expressed genes of P2RY12 were analyzed, followed by functional enrichment analysis. Furthermore, the correlation between P2RY12 and immune cell infiltrations was estimated using the TIMER database. P2RY12 expression was validated between 37 pairs of LUAD and normal tissues using RT-qPCR and immunohistochemistry. After overexpressing P2RY12, the proliferation and migration of A549 cells was detected by CCK-8 and scratch test.

RESULTS

145 up- and 102 down-regulated stromal- and immune-related mRNAs were identified for LUAD. Based on 145 up-regulated mRNAs, a PPI network was conducted, consisting of 95 nodes and 210 relationship pairs. Ten hub genes were then identified. Among them, CCR8, CNR2, CXCR5, GPR18, GPR31 and P2RY12 were in association with overall survival of patients with LUAD. After adjusting other variables, P2RY12 expression was an independent prognostic factor for LUAD. 288 co-expressed genes of P2RY12 were determined and these co-expressed genes were primarily involved in immune-related biological processes or pathways. Moreover, P2RY12 was significantly correlated with M2 macrophage and dendritic cell infiltration. After validation, P2RY12 expression was significantly decreased in LUAD than normal tissues. Its overexpression distinctly suppressed proliferation and migration of A549 cells.

CONCLUSION

Our findings suggest that P2RY12 is an immune infiltration-related prognostic marker for LUAD.

摘要

目的

越来越多的证据强调了 P2RY12 在致癌作用中的临床意义，P2RY12 属于 G 蛋白偶联受体家族。本研究旨在探讨 P2RY12 与肺腺癌（LUAD）肿瘤免疫微环境之间的关系。

方法

基于 352 例来自癌症基因组图谱（TCGA）的 LUAD 样本，通过 ESTIMATE 算法估计每个样本的基质和免疫评分。对基质/免疫高分和低分组间进行差异表达分析。然后通过 STRING 数据库构建蛋白质-蛋白质相互作用（PPI）网络。利用单因素和多因素 Cox 回归分析筛选预后相关因素。分析 P2RY12 的共表达基因，并进行功能富集分析。进一步使用 TIMER 数据库评估 P2RY12 与免疫细胞浸润的相关性。使用 RT-qPCR 和免疫组织化学法验证 37 对 LUAD 和正常组织中 P2RY12 的表达。过表达 P2RY12 后，通过 CCK-8 和划痕实验检测 A549 细胞的增殖和迁移。

结果

确定了 145 个上调和 102 个下调的 LUAD 基质和免疫相关 mRNAs。基于 145 个上调的 mRNAs，构建了一个 PPI 网络，包含 95 个节点和 210 个关系对。然后确定了 10 个枢纽基因。其中，CCR8、CNR2、CXCR5、GPR18、GPR31 和 P2RY12 与 LUAD 患者的总生存期相关。调整其他变量后，P2RY12 表达是 LUAD 的独立预后因素。确定了 288 个 P2RY12 的共表达基因，这些共表达基因主要参与免疫相关的生物过程或途径。此外，P2RY12 与 M2 巨噬细胞和树突状细胞浸润显著相关。验证后，P2RY12 在 LUAD 中的表达明显低于正常组织。其过表达明显抑制 A549 细胞的增殖和迁移。