Chen Yung-Liang, Chueh Fu-Shin, Yang Jai-Sing, Hsueh Shu-Ching, Lu Chi-Cheng, Chiang Jo-Hua, Lee Ching-Sung, Lu Hsu-Feng, Chung Jing-Gung
Department of Medical Laboratory Science and Biotechnology, Yuanpei University, Hsinchu 300, Taiwan.
School of Pharmacy, China Medical University, Taichung 404, Taiwan.
Environ Toxicol. 2015 Jul;30(7):803-15. doi: 10.1002/tox.21959. Epub 2014 Jan 29.
Irinotecan HCl (CPT-11) is an anticancer prodrug, but there is no available information addressing CPT-11-inhibited leukemia cells in in vitro and in vivo studies. Therefore, we investigated the cytotoxic effects of CPT-11 in promyelocytic leukemia HL-60 cells and in vivo and tumor growth in a leukemia xenograft model. Effects of CPT-11 on HL-60 cells were determined using flow cytometry, immunofluorescence staining, comet assay, real-time PCR, and Western blotting. CPT-11 demonstrated a dose- and time-dependent inhibition of cell growth, induction of apoptosis, and cell-cycle arrest at G0/G1 phase in HL-60 cells. CPT-11 promoted the release of AIF from mitochondria and its translocation to the nucleus. Bid, Bax, Apaf-1, caspase-9, AIF, Endo G, caspase-12, ATF-6b, Grp78, CDK2, Chk2, and cyclin D were all significantly upregulated and Bcl-2 was down-regulated by CPT-11 in HL-60 cells. Induction of cell-cycle arrest by CPT-11 was associated with changes in expression of key cell-cycle regulators such as CDK2, Chk2, and cyclin D in HL-60 cells. To test whether CPT-11 could augment antitumor activity in vivo, athymic BALB/c(nu/nu) nude mice were inoculated with HL-60 cells, followed by treatment with either CPT-11. The treatments significantly inhibited tumor growth and reduced tumor weight and volume in the HL-60 xenograft mice. The present study demonstrates the schedule-dependent antileukemia effect of CPT-11 using both in vitro and in vivo models. CPT-11 could potentially be a promising agent for the treatment of promyelocytic leukemia and requires further investigation.
盐酸伊立替康（CPT - 11）是一种抗癌前体药物，但在体外和体内研究中，尚无关于CPT - 11抑制白血病细胞的可用信息。因此，我们研究了CPT - 11对早幼粒细胞白血病HL - 60细胞的细胞毒性作用以及在白血病异种移植模型中的体内肿瘤生长情况。使用流式细胞术、免疫荧光染色、彗星试验、实时聚合酶链反应和蛋白质免疫印迹法测定CPT - 11对HL - 60细胞的影响。CPT - 11在HL - 60细胞中表现出剂量和时间依赖性的细胞生长抑制、细胞凋亡诱导以及G0/G1期细胞周期阻滞。CPT - 11促进了凋亡诱导因子（AIF）从线粒体的释放及其向细胞核的转位。在HL - 60细胞中，Bid、Bax、凋亡蛋白酶激活因子-1（Apaf - 1）、半胱天冬酶-9、AIF、核酸内切酶G（Endo G）、半胱天冬酶-12、活化转录因子6β（ATF - 6b）、葡萄糖调节蛋白78（Grp78）、细胞周期蛋白依赖性激酶2（CDK2）、细胞周期检测点激酶2（Chk2）和细胞周期蛋白D均被CPT - 11显著上调，而Bcl - 2被下调。CPT - 11诱导的细胞周期阻滞与HL - 60细胞中关键细胞周期调节因子如CDK2、Chk2和细胞周期蛋白D表达的变化有关。为了测试CPT - 11是否能增强体内抗肿瘤活性，将HL - 60细胞接种到无胸腺BALB/c（nu/nu）裸鼠体内，随后用CPT - 11进行治疗。这些治疗显著抑制了HL - 60异种移植小鼠的肿瘤生长，并降低了肿瘤重量和体积。本研究使用体外和体内模型证明了CPT - 11的时间依赖性抗白血病作用。CPT - 11可能是一种有前景的早幼粒细胞白血病治疗药物，需要进一步研究。
