核因子-κB相关信号在肾透明细胞癌中与预后和药物敏感性的关系

An NF-κB-related signature associated with prognosis and drug sensitivity in clear cell renal cell carcinoma

目前尚缺乏准确的基于基因表达谱的分层策略来预测肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)的预后.核因子-κB(nuclear factor-κB,NF-κB)基因家族是一组在生理和病理状态下参与免疫反应和炎症反应的转录因子.我们利用基因集富集分析(Gene Set Enrichment Analysis,GSEA)对癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据集进行分析,鉴定了9个与预后相关的NF-κB相关基因(NF-κB-related genes,NRGs).采用LASSO和Cox回归构建了NF-κB相关评分(NF-κB-related score,NRS)模型,并通过国际癌症基因组联盟(International Cancer Genome Consortium,ICGC)数据集进行验证,从而预测ccRCC患者生存的新的生物标记物.使用R软件(版本4.0)进行相关分析,探究NRGs与总体生存期(overall survival,OS)、临床相关性、肿瘤免疫微环境以及对美国食品药品监督管理局(Food and Drug Administration,FDA)批准的抗癌药物的药物敏感性之间的关系,结果发现,NRS分数与肿瘤免疫微环境、致癌途径以及ccRCC中的免疫治疗和药物敏感性显著相关.最后,通过实时定量聚合酶链反应和蛋白质印迹验证786-O cc RCC和HK-2细胞中9个NRGs的表达,观察到这9个NRGs基因在肿瘤细胞786-O细胞和人近端肾小管上皮细胞HK-2细胞中呈现差异表达.通过过表达TLR9,行CCK8细胞活性实验和划痕实验,检测TLR9对于ccRCC化疗辅助用药环磷酰胺敏感性的影响.总之,本研究建立了一个NRS模型,为进一步研究利用NRS来预测ccRCC生存和治疗的分子生物标志物提供了理论基础.

Accurate stratification strategy based on the gene expression profile to predict the prognosis of clear cell renal cell carcinoma(ccRCC)is not available.Nuclear factor-κB(NF-κB)gene family is a group of transcription factors mainly participating in immune and inflammatory reactions in physiological and pathological conditions.Here,we have established a model to predict ccRCC prognosis and patient survival.Using gene set enrichment analysis(GSEA)on The Cancer Genome Atlas(TCGA)dataset,we identified 9 NF-κB-related genes(NRGs)associated with prognosis.We constructed an NF-κB-related score(NRS)model using LASSO and Cox regression,and validated it using the International Cancer Genome Consortium(ICGC)dataset to predict novel biomarkers for survival in ccRCC patients.Using R software(version 4.0),we conducted correlation analysis to explore the relationship between NRGs and overall survival(OS),clinical relevance,tumor immune microenvironment,and sensitivity to FDA(Food and Drug Administration)-approved anticancer drugs.The results revealed a significant correlation between NRS scores and tumor immune microenvironment,carcinogenic pathways,as well as immune therapy and drug sensitivity in ccRCC.Finally,the expression of 9 NRGs in 786-O ccRCC and HK-2 cells was validated through real-time quantitative polymerase chain reaction(RTqPCR)and the west blotting.Differential expression of these 9 NRGs genes was observed between tumor cells(786-O cells)and human proximal tubule epithelial cells(HK-2 cells).By overexpressing TLR9,CCK8 and cell scratch experiments were performed to detect the effect of TLR9 on the sensitivity of ccRCC to the adjuvant chemotherapy drug cyclophosphamide.In summary,we constructed an NRS model,which proposes a rationale for further validation of NRS as a molecular biomarker for predicting survival and therapeutic responses in ccRCC.