Yu Rui, Lu Yefen, Yu Ren, Xie Jianjun, Zhou Shoujun
Department of Biochemistry and Molecular Biology, Zhejiang Key Laboratory of Pathophysiology, Medical School of Ningbo University, Ningbo, People's Republic of China.
Department of Neurology, The Fifth Affiliated Hospital of Wenzhou Medical University, Lishui, People's Republic of China.
Cancer Manag Res. 2021 Feb 3;13:953-963. doi: 10.2147/CMAR.S265788. eCollection 2021.
Renal cell carcinoma (RCC) is a common urological system malignancy lack of effective therapeutic options. Upregulation of the Bcl-2 proteins was correlated with poor prognosis of RCC, suggesting that BH-3 mimetics may be a promising treatment option. ABT-263 is a BH3 mimetic that possesses anti-tumor effects. TW-37 is another inhibitor of Bcl-2 family protein with potential anti-tumor activities. However, since their effect as single agent is limited, combination treatment represents a strategy to improve the efficiency. We studied the ABT-263 in combination with TW-37 and analyzed the molecular mechanisms of action in RCC cells.
MTT and colony formation assays were used to measure the proliferation of RCC cells. Transwell assay was used to assay the migration and invasion of RCC cells. Cell cycle distribution and apoptosis were measured using the flow cytometry and apoptotic nucleosome assay, respectively. Western blotting was performed to measure the change of proteins. The anti-tumor effects of ABT-263, TW-37 and their combination were also evaluated .
Cotreatment of TW-37 and ABT-263 synergistically repressed the proliferation of RCC cells. TW-37 and ABT-263 also synergistically inhibited the migration and invasion of RCC cells It was also showed that TW-37 and ABT-263 synergistically induced cell cycle arrest at the G2/M phase. Furthermore, increased apoptosis was observed after exposure to TW-37 and ABT-263. Mechanism investigation showed that TW-37 and ABT-263 synergistically induced apoptosis via the mitochondrial pathway and relied on the activation of Bax and caspases. Furthermore, ERK signaling pathway activation was detected after treated with TW-37 and ABT-263. Finally, TW-37 and ABT-263 also synergistically repressed the growth of RCC cells in xenograft mice.
In summary, our data demonstrated that combined treatment with TW-37 and ABT-263 exhibited synergistic RCC cell death and this combination may be applied as an effective therapeutic strategy against RCC.
肾细胞癌（RCC）是泌尿系统常见的恶性肿瘤，缺乏有效的治疗选择。Bcl-2蛋白上调与RCC预后不良相关，提示BH-3模拟物可能是一种有前景的治疗选择。ABT-263是一种具有抗肿瘤作用的BH3模拟物。TW-37是另一种具有潜在抗肿瘤活性的Bcl-2家族蛋白抑制剂。然而，由于它们作为单一药物的效果有限，联合治疗是提高疗效的一种策略。我们研究了ABT-263与TW-37联合使用，并分析了其在RCC细胞中的分子作用机制。
采用MTT和集落形成试验检测RCC细胞的增殖。采用Transwell试验检测RCC细胞的迁移和侵袭能力。分别用流式细胞术和凋亡核小体试验检测细胞周期分布和凋亡情况。通过蛋白质免疫印迹法检测蛋白质的变化。还评估了ABT-263、TW-37及其联合用药的抗肿瘤效果。
TW-37与ABT-263联合处理可协同抑制RCC细胞的增殖。TW-37和ABT-263还协同抑制RCC细胞的迁移和侵袭。还表明TW-37和ABT-263协同诱导细胞周期阻滞于G2/M期。此外，暴露于TW-37和ABT-263后观察到凋亡增加。机制研究表明，TW-37和ABT-263通过线粒体途径协同诱导凋亡，并依赖于Bax和半胱天冬酶的激活。此外，用TW-37和ABT-263处理后检测到ERK信号通路激活。最后，TW-37和ABT-263还协同抑制了异种移植小鼠体内RCC细胞的生长。
总之，我们的数据表明，TW-37与ABT-263联合治疗可协同诱导RCC细胞死亡，这种联合治疗可能作为一种有效的RCC治疗策略。
