Department of Urology, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, People's Republic of China.
Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation, Sun Yat-sen Memorial Hospital, Sun Yat-sen University, Guangzhou, 510120, People's Republic of China.
Drug Des Devel Ther. 2021 Dec 16;15:5061-5074. doi: 10.2147/DDDT.S343718. eCollection 2021.
Sunitinib is the first-line targeted drug for the treatment of advanced renal cell carcinoma (RCC), but its therapeutic potential is limited by premature drug resistance. In an attempt to overcome this limitation, a sunitinib-resistant cell-derived xenograft (CDX) model of clear cell renal cell carcinoma (ccRCC) in vivo was constructed in this study. The molecular mechanism of drug resistance was analyzed using sequencing and bioinformatics tools.
First, mice were injected subcutaneously with tumor cells 786-O to create tumors and were simultaneously treated with sunitinib. After three consecutive passages, a drug-resistant xenograft model was obtained. Then, key pathways and genes were identified via second-generation sequencing of the tissue and the examination of differentially expressed genes (DEGs) with bioinformatics tools.
Analysis of sequencing data revealed that 646 DEGs were upregulated and 465 were downregulated in the drug-resistant tissues when compared with the sensitive tissues. GO showed that the DEGs were significantly enriched in angiogenesis, cell hypoxia response, and apoptosis. KEGG analysis demonstrated that the main pathways were PI3K-Akt, HIF-1, NF-kappa B, and MAPK. Modular analysis of the PPI network indicated that the GO and KEGG analyses of module 1 with the highest ranking were mainly related to ubiquitinase activity. Similarly, the GO and KEGG analyses of the top 10 hub genes were also chiefly linked to ubiquitinase activity. Then, comprehensive expression analysis of the hub genes, and finally, the genes BTRC and TRIM32 were identified, which were consistent in all observations.
In this study, through the construction of in vitro models and bioinformatics analysis, the important pathways and key genes related to ccRCC sunitinib resistance were discovered. Among them, ubiquitinase may play an important role in drug resistance and may be a potential therapeutic target and biomarker.
舒尼替尼是治疗晚期肾细胞癌（RCC）的一线靶向药物，但由于其早期耐药性，其治疗潜力受到限制。本研究构建了一种体内舒尼替尼耐药肾透明细胞癌（ccRCC）衍生的异种移植（CDX）模型，试图克服这一局限性。采用测序和生物信息学工具分析耐药的分子机制。
首先，将肿瘤细胞 786-O 皮下注射到小鼠体内，形成肿瘤，并同时给予舒尼替尼治疗。连续传代 3 次后，获得耐药异种移植模型。然后，通过组织二代测序和生物信息学工具检测差异表达基因（DEGs），鉴定关键通路和基因。
测序数据分析显示，耐药组织与敏感组织相比，有 646 个 DEGs 上调，465 个 DEGs 下调。GO 分析显示，DEGs 主要富集在血管生成、细胞缺氧反应和细胞凋亡。KEGG 分析表明，主要通路为 PI3K-Akt、HIF-1、NF-kappa B 和 MAPK。PPI 网络的模块分析表明，排名最高的模块 1 的 GO 和 KEGG 分析主要与泛素酶活性相关。同样，前 10 个 hub 基因的 GO 和 KEGG 分析也主要与泛素酶活性相关。然后，对 hub 基因进行综合表达分析，最终发现 BTRC 和 TRIM32 基因在所有观察中一致。
本研究通过构建体外模型和生物信息学分析，发现了与 ccRCC 舒尼替尼耐药相关的重要通路和关键基因。其中，泛素酶可能在耐药中发挥重要作用，可能成为潜在的治疗靶点和生物标志物。
