Division of Hematology-Oncology, Department of Medicine, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul 06351, Korea.
Division of Hematology-Oncology, Department of Internal Medicine, Keimyung University Dongsan Hospital, Daegu 42601, Korea.
Int J Mol Sci. 2020 Apr 18;21(8):2825. doi: 10.3390/ijms21082825.
Despite comprehensive genomic analyses, no targeted therapies are approved for bladder cancer. Here, we investigate whether a single and combination therapy with targeted agents exert antitumor effects on bladder cancer cells through genomic alterations using a three-dimensional (3D) high-throughput screening (HTS) platform. Seven human bladder cancer cell lines were used to screen 24 targeted agents. The effects of 24 targeted agents were dramatically different according to the genomic alterations of bladder cancer cells. BEZ235 (dual phosphatidylinositol-3-kinase (PI3K)/mammalian target of rapamycin (mTOR) inhibitor) showed antitumor effects against most cell lines, while AZD2014 (mTOR inhibitor) had an IC50 value lower than 2 μM in 5637, J82, and RT4 cell lines. AZD5363 (protein kinase B (AKT) inhibitor) exerted antitumor effects on 5637, J82, and 253J-BV cells. J82 cells (PI3KCA and mTOR mutations) were sensitive to AZD5363, AZD2014, and BEZ235 alone or in AZD5363/AZD2014 and AZD5363/BEZ235 combinations. Although all single drugs suppressed cell proliferation, the combination of drugs exhibited synergistic effects on cell viability and colony formation. The synergistic effects of the combination therapy on the PI3K/Akt/mTOR pathway, apoptosis, and EMT were evident in Western blotting. Thus, the 3D culture-based HTS platform could serve as a useful preclinical tool to evaluate various drug combinations.
尽管进行了全面的基因组分析，但没有针对膀胱癌的靶向治疗方法获批。在这里，我们通过三维（3D）高通量筛选（HTS）平台研究了针对膀胱癌细胞的单一和联合靶向治疗是否通过基因组改变发挥抗肿瘤作用。使用七种人膀胱癌细胞系筛选了 24 种靶向药物。24 种靶向药物的作用根据膀胱癌细胞的基因组改变而有显著差异。BEZ235（双重磷脂酰肌醇-3-激酶（PI3K）/哺乳动物雷帕霉素靶蛋白（mTOR）抑制剂）对大多数细胞系表现出抗肿瘤作用，而 AZD2014（mTOR 抑制剂）在 5637、J82 和 RT4 细胞系中的 IC50 值低于 2 μM。AZD5363（蛋白激酶 B（AKT）抑制剂）对 5637、J82 和 253J-BV 细胞发挥抗肿瘤作用。J82 细胞（PI3KCA 和 mTOR 突变）对 AZD5363、AZD2014 和 BEZ235 单独或 AZD5363/AZD2014 和 AZD5363/BEZ235 联合治疗敏感。虽然所有单一药物均抑制细胞增殖，但药物联合治疗对细胞活力和集落形成具有协同作用。联合治疗对 PI3K/Akt/mTOR 通路、细胞凋亡和 EMT 的协同作用在 Western blot 中明显。因此，基于 3D 培养的 HTS 平台可以作为评估各种药物组合的有用临床前工具。
