Lv Yinxiang, Liu Rongrong, Xie Shangzhi, Zheng Xiaoxiao, Mao Jiayan, Cai Ying, Chen Wei
aDepartment of Oncology, Xinchang People's Hospital, Shaoxing bDepartment of Hematology-Oncology, Children's Hospital cThe Second Affiliated Hospital, School of Medicine, Zhejiang University, Zhejiang dMeizhong Disease Gene Research Institute Company Limited, Meizhong, People's Repubic of China.
Anticancer Drugs. 2017 Sep;28(8):861-868. doi: 10.1097/CAD.0000000000000527.
Calcein acetoxymethyl ester (calcein-AM) treatment has been reported to exert antitumor effects in certain cancer cells; however, the detailed mechanism of action of calcein-AM in cancers remains unclear, especially in nonsmall cell lung cancer (NSCLC). This study focused on the function and mechanism of action of calcein-AM in NSCLC. We used cell viability assays, western blotting, and EdU proliferation assay combined with calcein-AM treatment or siRNA interference to investigate the role of topoisomerase IIβ binding protein 1 (TopBP1) and p53 in NSCLC chemotherapy. We found that calcein-AM has antitumor effects in lung cancer and enhances the antitumor effects of doxorubicin in NSCLC. Furthermore, we found that TopBP1, which we previously showed was involved in doxorubicin resistance through upregulation of aberrant p53, was involved in calcein-AM-mediated increased doxorubicin sensitivity. Doxorubicin upregulated the expression of aberrant p53. Calcein-AM repressed the expression of TopBP1, which resulted in reduced expression of aberrant p53 and disrupted the antiapoptotic activity mediated by the TopBP1/mutp53 pathway in NSCLC. Together, our findings show that calcein-AM, the cell-permeable derivative of calcein, exerts significant antitumor effects in NSCLC, and can enhance the antitumor effect of doxorubicin by regulating the TopBP1/mutp53 pathway. These findings provide novel insight into lung cancer treatment.
据报道，钙黄绿素乙酰氧基甲酯（calcein-AM）处理在某些癌细胞中具有抗肿瘤作用；然而，calcein-AM在癌症中的详细作用机制仍不清楚，尤其是在非小细胞肺癌（NSCLC）中。本研究聚焦于calcein-AM在NSCLC中的功能及作用机制。我们使用细胞活力测定、蛋白质印迹法以及EdU增殖测定，并结合calcein-AM处理或小干扰RNA（siRNA）干扰，来研究拓扑异构酶IIβ结合蛋白1（TopBP1）和p53在NSCLC化疗中的作用。我们发现calcein-AM在肺癌中具有抗肿瘤作用，并增强了阿霉素在NSCLC中的抗肿瘤作用。此外，我们发现，我们之前表明通过异常p53的上调参与阿霉素耐药的TopBP1，也参与了calcein-AM介导的阿霉素敏感性增加。阿霉素上调了异常p53的表达。Calcein-AM抑制了TopBP1的表达，这导致NSCLC中异常p53的表达降低，并破坏了由TopBP1/突变型p53途径介导的抗凋亡活性。总之，我们的研究结果表明，calcein的细胞可渗透衍生物calcein-AM在NSCLC中发挥显著的抗肿瘤作用，并可通过调节TopBP1/突变型p53途径增强阿霉素的抗肿瘤作用。这些发现为肺癌治疗提供了新的见解。
