RGS17通过激活cAMP信号通路促进肾透明细胞癌进展

RGS17 promotes the progression of kidney renal clear cell carcinoma via activating the cAMP signaling pathway

摘要

目的 探讨G蛋白信号蛋白家族的调节因子17（regulator of G protein signaling 17，RGS17）在肾透明细胞癌患者中的临床意义和功能机制。 方法 获得癌症基因组图谱（The Cancer Genome Atlas，TCGA）数据库中肾透明细胞癌（kidney renal clear cell carcinoma，KIRC）的RNAseq数据和相应的临床信息。利用R软件研究RGS17在KIRC中的表达差异及其与临床特征的关系。使用免疫组化及PCR进行验证。采用Kaplan-Meier（K-M）分析、受试者工作特征（receiver operating characteristic，ROC）曲线、单因素COX分析、多因素COX分析来评估患者的生存和预后，并构建nomogram模型。使用STRING进行RGS17相关基因的PPI网络构建，并进行 GO及KEGG富集分析。并采用 RT-qPCR、Western blot、CCK-8、Transwell 和 划痕实验等方法检测RGS17对ACHN细胞增殖迁移及cAMP信号通路的影响。 结果 RGS17在KIRC中高表达，且RGS17的高表达与更高的T分期、临床分期、肿瘤转移显著相关。K-M生存分析显示，RGS17上调与KIRC患者总生存期、无疾病进展生存期下降密切相关。ROC曲线表明RGS17能较好的区分正常和KIRC患者，并在预测KIRC患者的预后方面具有一定的准确性。RGS17是KIRC独立预后因素。使用RGS17表达、临床分期、病理分级构建nomogram预后模型能较好预测1、3、5年生存率。RGS17敲低显著抑制ACHN细胞的增殖、迁移和侵袭能力。另外，RGS17敲低能够抑制cAMP信号通路的激活。 结论 RGS17在肾透明细胞癌中具有促癌基因的作用，其可能通过激活cAMP信号通路促进肾透明细胞癌的进展。

Abstract

Objective To investigate the clinical significance and functional mechanism of regulator of G protein signaling 17 （RGS17） in patients with kidney renal clear cell carcinoma（KIRC）. Methods RNAseq data and related clinical information on KIRC were obtained from The Cancer Genome Atlas（TCGA） database. The differential expression of RGS17 in KIRC and its relationship with clinical characteristics were studied using R software. and the results were validated using immunohistochemistry and polymerase chain reaction（PCR）. Patient survival and outcome were analyzed by using the Kaplan-Meier（K-M） method，receiver operating characteristic（ROC） curve，and univariable and multivariable Cox analyses，and a nomogram model was constructed. We constructed the protein-protein interaction（PPI） network of RGS17-related genes using the STRING database，and performed Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses. The effects of RGS17 on the proliferation and migration of ACHN cells and the cAMP signaling pathway were determined by real-time quantitative PCR，Western blot，Cell Counting Kit-8，transwell assay，and wound healing assay. Results RGS17 was highly expressed in KIRC，and its high expression was significantly associated with a higher T stage，a higher clinical stage，and tumor metastasis. The K-M survival analysis showed that RGS17 upregulation was closely related to decreased overall survival and progression-free survival in patients with KIRC. The ROC curve analysis indicated that RGS17 could well distinguish between normal people and patients with KIRC，showing certain accuracy in predicting the outcome of patients with KIRC. RGS17 was identified as an independent prognostic factor for KIRC. The nomogram model constructed using RGS17 expression，clinical stage，and pathological grade could well predict the 1-，3-，and 5-year survival rates. Knocking RGS17 down significantly suppressed the proliferation，migration，and invasion abilities of ACHN cells as well as the activation of the cAMP signaling pathway. Conclusion RGS17 acts as an oncogene in KIRC，which may promote the progression of KIRC by activating the cAMP signaling pathway.

关键词

肾透明细胞癌 / G蛋白信号蛋白家族的调节因子17 / 细胞增殖 / 细胞迁移和侵袭 / PKA/CREB信号通路

Key words

clear cell renal cell carcinoma / regulator of G protein signaling 17 / cell proliferation / cell migration / invasion / PKA/CREB signaling pathway

基金资助

2021年医疗卫生领域指导性科技计划资助项目(泉科〔2021〕150号)