Department of Nephrology, The First Affiliated Hospital, Jiangxi Medical College, Nanchang University, Nanchang, 330000, Jiangxi Province, China.
Key Laboratory of Urinary System Diseases of Jiangxi Province, Nanchang, China.
Sci Rep. 2024 Aug 30;14(1):20204. doi: 10.1038/s41598-024-69892-5.
Numerous studies have shown that the Krüppel-like factors (KLFs) family of transcription factors regulate various eukaryotic physiological processes including the proliferation, differentiation, senescence, death, and carcinogenesis of animal cells. In addition, they are involved in the regulation of key biological processes such as cell cycle, DNA repair, and immune response. Current studies focus on investigating the role of KLFs in normal physiological conditions and the incidence and development of diseases. However002C the significance of KLFs family genes in clear cell renal cell carcinoma (ccRCC) remains partly understood; therefore, an in-depth investigation of their role and clinical value in this cancer is desired. The study aimed to investigate the role of KLF family genes in the incidence, development, and prognosis of ccRCC, and to identify the related potential biomarkers and therapeutic targets. The expression of KLFs in the RNA sequencing data of 613 ccRCCs from the TCGA database was analyzed using R software, and UALCAN and GEPIA assessed the expression of KLF genes in ccRCC. Real-time fluorescence quantitative PCR analysis was performed using 10 pairs of paired ccRCC sample tissues and renal cancer cell lines from the First Affiliated Hospital of Nanchang University. Overall survival (OS), progression-free interval (PFI), and disease-specific survival (DSS) of Kidney Clear Cell Carcinoma (KIRC) samples at differential expressions of KLFs in the TCGA database were analyzed using the R software, followed by generating a nomogram prediction model. GSCALite assessed the interactions of KLF genes with miRNAs and generated network maps. Protein interaction network maps of 50 neighboring genes associated with KLF mutations were analyzed using STRING with GO and KEGG functional enrichment analyses. The cBioPortal determined the probability of KLF gene mutations and their impact on OS and disease-free survival (DFS) in patients with ccRCC. Immune cell infiltration of KLFs was analyzed using TIMER. Finally, GSCALite was used to analyze the drug sensitivity and associated pathways of action of KLFs. Correlation validation using cellular experiments. KLF3/5/9/15 were significantly downregulated in ccRCC tissues, whereas KLF16/17 were upregulated compared with the adjacent tissues. Patients with high mRNA levels of KLF16/17 showed significantly lower OS, PFI, and DSS, whereas KLF3/5/9 showed a reverse trend. In patients with ccRCC, a significant correlation was observed between KLF mutations and OS and DSS. Furthermore, the correlation of KLF3/5/9 with immune cell infiltration was stronger than that of KLF15/16, while KLF17 was significantly associated with the Epithelial-Mesenchymal Transition (EMT) pathway. Overexpression of KLF5 inhibits the proliferative and migratory capacity of renal cancer cells (786-O and OS-RC-2), as well as their sensitivity to relevant small molecule drugs. Our research revealed the expression levels and biological significance of KLF genes in ccRCC, particularly highlighting the potential of KLF5 as a promising biomarker and therapeutic target for effective prognosis and diagnosis of ccRCC.
大量研究表明,Krüppel 样因子(KLF)转录因子家族调节包括动物细胞增殖、分化、衰老、死亡和癌发生在内的各种真核生理过程。此外,它们参与细胞周期、DNA 修复和免疫反应等关键生物过程的调节。目前的研究重点是调查 KLF 在正常生理条件下和疾病的发生和发展中的作用。然而,KLF 家族基因在透明细胞肾细胞癌(ccRCC)中的意义部分仍未被理解;因此,需要深入研究它们在这种癌症中的作用和临床价值。本研究旨在探讨 KLF 家族基因在 ccRCC 发病、发展和预后中的作用,以及鉴定相关的潜在生物标志物和治疗靶点。使用 R 软件分析了 TCGA 数据库中 613 例 ccRCC 的 RNA 测序数据中 KLFs 的表达情况,UALCAN 和 GEPIA 评估了 ccRCC 中 KLF 基因的表达。使用来自南昌大学第一附属医院的 10 对配对的 ccRCC 样本组织和肾癌细胞系进行实时荧光定量 PCR 分析。使用 R 软件分析 TCGA 数据库中 KLFs 差异表达的 KIRC 样本的总生存(OS)、无进展间隔(PFI)和疾病特异性生存(DSS),并生成预测模型。GSCALite 评估了 KLF 基因与 miRNAs 的相互作用,并生成了网络图谱。使用 STRING 进行与 KLF 突变相关的 50 个邻近基因的蛋白质相互作用网络图谱的分析,并进行 GO 和 KEGG 功能富集分析。cBioPortal 确定了 KLF 基因突变的概率及其对 ccRCC 患者 OS 和无病生存(DFS)的影响。使用 TIMER 分析 KLFs 的免疫细胞浸润情况。最后,使用 GSCALite 分析 KLFs 的药物敏感性及其相关作用途径。通过细胞实验进行相关性验证。KLF3/5/9/15 在 ccRCC 组织中显著下调,而 KLF16/17 则上调。与相邻组织相比,KLF16/17 mRNA 水平高的患者 OS、PFI 和 DSS 明显降低,而 KLF3/5/9 则呈现相反的趋势。在 ccRCC 患者中,KLF 突变与 OS 和 DSS 之间存在显著相关性。此外,KLF3/5/9 与免疫细胞浸润的相关性强于 KLF15/16,而 KLF17 与上皮-间充质转化(EMT)途径显著相关。KLF5 的过表达抑制肾癌细胞(786-O 和 OS-RC-2)的增殖和迁移能力,以及它们对相关小分子药物的敏感性。我们的研究揭示了 KLF 基因在 ccRCC 中的表达水平和生物学意义,特别强调了 KLF5 作为一种有前途的生物标志物和治疗靶点,可有效预测和诊断 ccRCC。