Department of Neurosurgery, Zhongnan Hospital, Wuhan University, Wuhan, China.
Department of Urology, Zhongnan Hospital of Wuhan University, Wuhan, China.
J Cancer Res Clin Oncol. 2023 Oct;149(13):11057-11071. doi: 10.1007/s00432-023-04983-w. Epub 2023 Jun 20.
Patients with clear cell renal cell carcinoma (ccRCC), which is the most commonly diagnosed subtype of renal cell carcinoma, are at risk of tumor metastasis and recrudescence. Previous research has shown that oxidative stress can induce tumorigenesis in many cancers and can be a target of cancer treatment. Despite these findings, little progress has been made understanding in the association of oxidative stress-related genes (OSRGs) with ccRCC.
In vitro experiments were conducted with MTT survival assays, qRT‒PCR, apoptosis assays, cell cycle assays, ROS assays, and IHC staining.
In our study, 12 differentially expressed oxidative stress-related genes (DEOSGs) and related transcription factors (TFs) that are relevant to overall survival (OS) were screened, and their mutual regulatory networks were constructed with data from the TCGA database. Moreover, we constructed a risk model of these OSRGs and performed clinical prognostic analysis and validation. Next, we performed protein-protein interaction (PPI) network analysis and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of MELK, PYCR1, and PML. A tissue microarray also verified the high expression of MELK and PYCR1 in ccRCC. Finally, in vitro cellular experiments demonstrated that knockdown of MELK or PYCR1 significantly inhibited ccRCC cell proliferation by causing cell apoptosis and inducing cell cycle arrest in the G1 phase. Intracellular ROS levels were elevated after these two genes were knocked down.
Our results revealed the potential DEORGs to be used in ccRCC prognostic prediction and identified two biomarkers, named PYCR1 and MELK, which regulated the proliferation of ccRCC cells by affecting ROS levels. Furthermore, PYCR1 and MELK could be promising targets for predicting the progression and prognosis of ccRCC, thereby serving as new targets for medical treatments.
透明细胞肾细胞癌（ccRCC）是最常见的肾细胞癌亚型，患者存在肿瘤转移和复发的风险。既往研究表明，氧化应激可诱发多种癌症的发生，且可能成为癌症治疗的靶点。尽管已有这些发现，但人们对氧化应激相关基因（OSRGs）与 ccRCC 之间的关联仍知之甚少。
采用 MTT 生存分析、qRT-PCR、凋亡分析、细胞周期分析、ROS 分析和 IHC 染色进行体外实验。
在本研究中，我们筛选了 12 个差异表达的氧化应激相关基因（DEOSGs）和相关转录因子（TFs），这些基因与总生存期（OS）相关，并利用 TCGA 数据库中的数据构建了它们的互调控网络。此外，我们构建了这些 OSRGs 的风险模型，并进行了临床预后分析和验证。接下来，我们进行了蛋白质-蛋白质相互作用（PPI）网络分析以及 MELK、PYCR1 和 PML 的基因本体（GO）和京都基因与基因组百科全书（KEGG）通路富集分析。组织微阵列还验证了 MELK 和 PYCR1 在 ccRCC 中的高表达。最后，体外细胞实验表明，敲低 MELK 或 PYCR1 可通过诱导细胞凋亡和使细胞周期停滞在 G1 期，显著抑制 ccRCC 细胞的增殖。敲低这两个基因后，细胞内 ROS 水平升高。
我们的研究结果揭示了用于 ccRCC 预后预测的潜在 DEORGs，并确定了两个标志物，即 PYCR1 和 MELK，它们通过影响 ROS 水平来调节 ccRCC 细胞的增殖。此外，PYCR1 和 MELK 可能成为预测 ccRCC 进展和预后的有前途的靶点，从而成为新的治疗靶点。
