Gupta Pranav, Zhang Yun-Kai, Zhang Xiao-Yu, Wang Yi-Jun, Lu Kimberly W, Hall Timothy, Peng Richard, Yang Dong-Hua, Xie Ni, Chen Zhe-Sheng
Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John's University, Queens, New York, USA.
Institute of translation medicine, Shenzhen Second People's Hospital, First Affiliated Hospital of Shenzhen University, Shenzhen, China.
Cell Physiol Biochem. 2018;45(4):1515-1528. doi: 10.1159/000487578. Epub 2018 Feb 19.
BACKGROUND/AIMS: The overexpression of ATP-Binding Cassette (ABC) transporters has known to be one of the major obstacles impeding the success of chemotherapy in drug resistant cancers. In this study, we evaluated voruciclib, a CDK 4/6 inhibitor, for its chemo-sensitizing activity in ABCB1- and ABCG2- overexpressing cells.
Cytotoxicity and reversal effect of voruciclib was determined by MTT assay. The intracellular accumulation and efflux of ABCB1 and ABCG2 substrates were measured by scintillation counter. The effects on expression and intracellular localization of ABCB1 and ABCG2 proteins were determined by Western blotting and immunofluorescence, respectively. Vanadate-sensitive ATPase assay was done to determine the effect of voruciclib on the ATPase activity of ABCB1 and ABCG2. Flow cytometric analysis was done to determine the effect of voruciclib on apoptosis of ABCB1 and ABCG2-overexpressing cells and docking analysis was done to determine the interaction of voruciclib with ABCB1 and ACBG2 protein.
Voruciclib significantly potentiated the effect of paclitaxel and doxorubicin in ABCB1-overexpressing cells, as well as mitoxantrone and SN-38 in ABCG2-overexpressing cells. Voruciclib moderately sensitized ABCC10- overexpressing cells to paclitaxel, whereas it did not alter the cytotoxicity of substrates of ABCC1. Furthermore, voruciclib increased the intracellular accumulation and decreased the efflux of substrate anti-cancer drugs from ABCB1- or ABCG2-overexpressing cells. However, voruciclib did not alter the expression or the sub-cellular localization of ABCB1 or ABCG2. Voruciclib stimulated the ATPase activity of both ABCB1 and ABCG2 in a concentration-dependent manner. Lastly, voruciclib exhibited a drug-induced apoptotic effect in ABCB1- or ABCG2- overexpressing cells.
Voruciclib is currently a phase I clinical trial drug. Our findings strongly support its potential use in combination with conventional anti-cancer drugs for cancer chemotherapy.
背景/目的：已知ATP结合盒（ABC）转运蛋白的过表达是阻碍耐药性癌症化疗成功的主要障碍之一。在本研究中，我们评估了CDK 4/6抑制剂沃鲁西利布在ABCB1和ABCG2过表达细胞中的化疗增敏活性。
通过MTT法测定沃鲁西利布的细胞毒性和逆转作用。用闪烁计数器测量ABCB1和ABCG2底物的细胞内积累和外排。分别通过蛋白质免疫印迹法和免疫荧光法测定对ABCB1和ABCG2蛋白表达和细胞内定位的影响。进行钒酸盐敏感ATP酶测定以确定沃鲁西利布对ABCB1和ABCG2的ATP酶活性的影响。通过流式细胞术分析来确定沃鲁西利布对ABCB1和ABCG2过表达细胞凋亡的影响，并进行对接分析以确定沃鲁西利布与ABCB1和ACBG2蛋白的相互作用。
沃鲁西利布显著增强了紫杉醇和阿霉素在ABCB1过表达细胞中的作用，以及米托蒽醌和SN-38在ABCG2过表达细胞中的作用。沃鲁西利布使ABCC10过表达细胞对紫杉醇有适度的敏感性，而它没有改变ABCC1底物的细胞毒性。此外，沃鲁西利布增加了ABCB1或ABCG2过表达细胞中底物抗癌药物的细胞内积累并减少了其外排。然而，沃鲁西利布没有改变ABCB1或ABCG2的表达或亚细胞定位。沃鲁西利布以浓度依赖性方式刺激ABCB1和ABCG2的ATP酶活性。最后，沃鲁西利布在ABCB1或ABCG2过表达细胞中表现出药物诱导的凋亡作用。
沃鲁西利布目前是一种I期临床试验药物。我们的研究结果有力地支持了其与传统抗癌药物联合用于癌症化疗的潜在用途。
