Institute of Gansu Nephro-Urological Clinical Center, Department of Urology, Institute of Urology, Key Laboratory of Urological Disease of Gansu Province, Lanzhou University Second Hospital, Lanzhou, China.
J Cell Biochem. 2019 Apr;120(4):6347-6360. doi: 10.1002/jcb.27922. Epub 2018 Oct 10.
Chemotherapy is still a standard treatment of unresectable bladder cancer or distant metastases. The chemotherapy resistance always occurs after a period of treatment indicating poor prognosis. The current study aimed to explore the molecular mechanism of chemoresistance in bladder cancer cells. The gene expression profiles of GSE77883, including three untreated T24 cells samples and three gemcitabine-resistant T24 cells samples, was downloaded from Gene Expression Omnibus database. The screening of differentially expressed genes (DEGs), gene function analysis, and interaction prediction between microRNAs (miRNAs) and DEGs were performed by R software. The protein-protein interaction (PPI) and miRNA-DEGs networks were constructed and visualized by Cytoscape software. Then, the small molecules, with potential synergistic or antagonistic effects to gemcitabine resistance, were identified using the Connectivity Map database. Finally, gemcitabine-resistant T24 cell line was established and key genes were validated by quantitative real-time polymerase chain reaction (qRT-PCR). In total, 536 upregulated and 513 downregulated genes were screened and mainly enriched in oxidative stress response and signaling pathways related to extracellular matrix-receptor interaction and focal adhesion. PPI network showed interleukin 6, tumor necrosis factor, kinesin family member 11, and BUB1 mitotic checkpoint serine/threonine kinase B were key genes. The miRNA-DEGs regulatory networks included 18 miRNAs and 185 DEGs, including miR-182-5p, miR-590-3p, miR-320a and serum- and glucocorticoid-regulated kinase 1 (SGK1). Then, the related key genes and miRNAs were confirmed by qRT-PCR. Furthermore, 81 small molecules with antagonistic or synergistic effect to GEM were screened. We have investigated the molecular mechanisms driving GEM-resistance in bladder cancer cells that would contribute to the development of chemotherapy for advanced bladder cancer.
化疗仍然是不可切除的膀胱癌或远处转移的标准治疗方法。在经过一段时间的治疗后，化疗耐药总是会发生，预示着预后不良。本研究旨在探索膀胱癌细胞化疗耐药的分子机制。从基因表达综合数据库中下载了 GSE77883 的基因表达谱，包括三个未经处理的 T24 细胞样本和三个吉西他滨耐药的 T24 细胞样本。使用 R 软件进行差异表达基因（DEGs）的筛选、基因功能分析和 miRNA-DEGs 之间的相互作用预测。使用 Cytoscape 软件构建和可视化蛋白质-蛋白质相互作用（PPI）和 miRNA-DEGs 网络。然后，使用 Connectivity Map 数据库鉴定具有潜在协同或拮抗吉西他滨耐药作用的小分子。最后，通过定量实时聚合酶链反应（qRT-PCR）建立吉西他滨耐药 T24 细胞系并验证关键基因。共筛选出 536 个上调和 513 个下调基因，主要富集在氧化应激反应和细胞外基质-受体相互作用及焦点黏附相关信号通路中。PPI 网络显示白细胞介素 6、肿瘤坏死因子、驱动蛋白家族成员 11 和 BUB1 有丝分裂检查点丝氨酸/苏氨酸激酶 B 是关键基因。miRNA-DEGs 调控网络包括 18 个 miRNA 和 185 个 DEGs，其中包括 miR-182-5p、miR-590-3p、miR-320a 和血清和糖皮质激素调节激酶 1（SGK1）。然后，通过 qRT-PCR 验证了相关的关键基因和 miRNA。此外，筛选出 81 种对 GEM 具有拮抗或协同作用的小分子。我们已经研究了驱动膀胱癌细胞中 GEM 耐药的分子机制，这将有助于开发晚期膀胱癌的化疗。
