综合分析鉴定 CCT3 为调节因子，可重塑肺腺癌免疫抑制性肿瘤微环境。

Integrated analyzes identify CCT3 as a modulator to shape immunosuppressive tumor microenvironment in lung adenocarcinoma.

机构信息

Department of Laboratory Medicine, Second Hospital of Anhui Medical University, Hefei, China.

Research Center for Translational Medicine, The Second Hospital of Anhui Medical University, Hefei, China.

出版信息

BMC Cancer. 2023 Mar 14;23(1):241. doi: 10.1186/s12885-023-10677-w.

DOI:10.1186/s12885-023-10677-w

PMID:36918801

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC10012614/

Abstract

BACKGROUND

Chaperonin-containing tailless complex polypeptide 1 (TCP1) subunit 3 (CCT3) has tumor-promoting effects in lung adenocarcinoma (LUAD). This study aims to investigate the molecular mechanisms of CCT3 in LUAD oncogenesis.

METHODS

The UALCAN databases, Human Protein Atlas (HPA) and The Cancer Genome Atlas (TCGA) data were used to analyze CCT3 expression in LUAD. Both the Wilcoxon rank-sum test and the regression model were used to investigate the connection between clinicopathologic characteristics of LUAD patients and CCT3 expression. The prognostic value of CCT3 was determined by Cox regression models, the Kaplan-Meier method and Nomogram prediction. Next, we identified the most related genes with CCT3 via GeneMANIA and String databases, and the association between CCT3 and infiltrated immune cells using single-sample Gene Set Enrichment Analysis (ssGSEA). CCT3-related pathway enrichment analysis was investigated by GSEA. Finally, CCT3 roles in cell proliferation and apoptosis of LUAD A549 cells was verified by siRNA (small interfering RNA) mediated CCT3 knockdown.

RESULTS

CCT3 was upregulated in LUAD both in mRNA and protein levels. CCT3 overexpression was associated with clinicopathological characteristics including sex, smoking, T- and N-categories, pathological staging, and a poor prognosis of LUAD patients. GeneMANIA and String databases found a set of CCT3-related genes that are connected to the assembly and stability of proteins involved in proteostasis of cytoskeletal filaments, DNA repair and protein methylation. Furthermore, CCT3 was found to be positively correlated with the infiltrating Th2 cells (r = 0.442, p < 0.01) while negatively correlated with mast cells (r = -0.49, p < 0.01) and immature dendritic cells (iDCs, r = -0.401, p < 0.001) according to ssGSEA analyzes. The pathway analysis based on GSEA method showed that the cell cycle pathway, the protein export pathway, the proteasome pathway and the ribosome pathway are enriched in CCT3 high group, whereas the JAK/STAT pathway, B cell receptor pathway, T cell receptor pathway and toll like receptor pathway were enriched in CCT3 low group. Finally, CCT3 knockdown substantially inhibited proliferation while promoted apoptosis of A549 cells.

CONCLUSION

Integrated analyzes identify CCT3 as a modulator to shape immunosuppressive tumor microenvironment in LUAD and therefore, a prognostic factor for LUAD.

摘要

背景

含伴侣素的无尾复合物多肽 1（TCP1）亚基 3（CCT3）在肺腺癌（LUAD）中具有促进肿瘤的作用。本研究旨在探讨 CCT3 在 LUAD 致癌中的分子机制。

方法

使用 UALCAN 数据库、人类蛋白质图谱（HPA）和癌症基因组图谱（TCGA）数据分析 LUAD 中 CCT3 的表达。采用 Wilcoxon 秩和检验和回归模型分析 LUAD 患者的临床病理特征与 CCT3 表达的关系。采用 Cox 回归模型、Kaplan-Meier 方法和 Nomogram 预测确定 CCT3 的预后价值。接下来，我们通过 GeneMANIA 和 String 数据库确定与 CCT3 最相关的基因，并通过单样本基因集富集分析（ssGSEA）确定 CCT3 与浸润免疫细胞的关系。通过 GSEA 进行 CCT3 相关通路富集分析。最后，通过 siRNA（小干扰 RNA）介导的 CCT3 敲低验证 CCT3 在 LUAD A549 细胞增殖和凋亡中的作用。

结果

CCT3 在 LUAD 的 mRNA 和蛋白水平均上调。CCT3 过表达与临床病理特征有关，包括性别、吸烟、T 期和 N 期、病理分期和 LUAD 患者的不良预后。GeneMANIA 和 String 数据库发现了一组与 CCT3 相关的基因，这些基因与参与细胞骨架丝蛋白组装和稳定性、DNA 修复和蛋白质甲基化的蛋白质的组装和稳定性有关。此外，根据 ssGSEA 分析，CCT3 与浸润性 Th2 细胞呈正相关（r=0.442，p<0.01），而与肥大细胞（r=-0.49，p<0.01）和不成熟树突状细胞（iDCs，r=-0.401，p<0.001）呈负相关。基于 GSEA 方法的通路分析表明，细胞周期通路、蛋白质输出通路、蛋白酶体通路和核糖体通路在 CCT3 高组中富集，而 JAK/STAT 通路、B 细胞受体通路、T 细胞受体通路和 Toll 样受体通路在 CCT3 低组中富集。最后，CCT3 敲低显著抑制了 A549 细胞的增殖，同时促进了其凋亡。