DataSheet1_Transcription Factors BARX1 and DLX4 Contribute to Progression of Clear Cell Renal Cell Carcinoma via Promoting Proliferation and Epithelial–Mesenchymal Transition.docx

Dysregulation of transcription factors contributes to the carcinogenesis and progression of cancers. However, their roles in clear cell renal cell carcinoma remain largely unknown. This study aimed to evaluate the clinical significance of TFs and investigate their potential molecular mechanisms in ccRCC. Data were accessed from the cancer genome atlas kidney clear cell carcinoma cohort. Bioinformatics algorithm was used in copy number alterations mutations, and differentially expressed TFs’ analysis. Univariate and multivariate Cox regression analyses were performed to identify clinically significant TFs and construct a six-TF prognostic panel. TFs’ expression was validated in human tissues. Gene set enrichment analysis (GSEA) was utilized to find enriched cancer hallmark pathways. Functional experiments were conducted to verify the cancer-promoting effect of BARX homeobox 1 (BARX1) and distal-less homeobox 4 (DLX4) in ccRCC, and Western blot was performed to explore their downstream pathways. As for results, many CNAs and mutations were identified in transcription factor genes. TFs were differentially expressed in ccRCC. An applicable predictive panel of six-TF genes was constructed to predict the overall survival for ccRCC patients, and its diagnostic efficiency was evaluated by the area under the curve (AUC). BARX1 and DLX4 were associated with poor prognosis, and they could promote the proliferation and migration of ccRCC. In conclusion, the six-TF panel can be used as a prognostic biomarker for ccRCC patients. BARX1 and DLX4 play oncogenic roles in ccRCC via promoting proliferation and epithelial–mesenchymal transition. They have the potential to be novel therapeutic targets for ccRCC.

转录因子（transcription factor, TF）的表达失调参与了癌症的发生与进展过程，但其在透明细胞肾细胞癌（clear cell renal cell carcinoma, ccRCC）中的具体作用仍未被充分阐明。本研究旨在评估转录因子在透明细胞肾细胞癌中的临床价值，并探究其在该疾病中潜在的分子调控机制。本研究数据来源于癌症基因组图谱（The Cancer Genome Atlas, TCGA）透明细胞肾细胞癌队列。本研究采用生物信息学算法开展拷贝数变异（copy number alteration, CNA）、突变及差异表达转录因子的相关分析。通过单因素及多因素Cox回归分析，筛选出具有临床意义的转录因子，并构建了由6个转录因子组成的预后特征模型。研究通过人体组织样本验证了上述转录因子的表达水平。采用基因集富集分析（Gene Set Enrichment Analysis, GSEA）筛选出显著富集的癌症特征通路。通过功能实验验证了BARX同源框1（BARX homeobox 1, BARX1）及远端同源框4（distal-less homeobox 4, DLX4）在透明细胞肾细胞癌中的促癌作用，并采用蛋白质印迹法（Western blot）探究了二者的下游调控通路。研究结果显示，转录因子基因中存在大量拷贝数变异与突变事件，且透明细胞肾细胞癌组织中存在差异表达的转录因子。本研究构建了一套可用于预测透明细胞肾细胞癌患者总生存期的六转录因子基因预后特征模型，并通过曲线下面积（Area Under the Curve, AUC）评估了该模型的诊断效能。BARX1与DLX4的异常高表达与患者不良预后显著相关，且二者可促进透明细胞肾细胞癌细胞的增殖与迁移能力。综上，六转录因子预后特征模型可作为透明细胞肾细胞癌患者的预后生物标志物。BARX1与DLX4通过促进细胞增殖与上皮间质转化（epithelial-mesenchymal transition, EMT）在透明细胞肾细胞癌中发挥致癌作用，二者有望成为该疾病的新型治疗靶点。