Antitumor effects with apoptotic death in human promyelocytic leukemia HL-60 cells and suppression of leukemia xenograft tumor growth by irinotecan HCl.
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.
Department of Medical Laboratory Science and Biotechnology, Yuanpei University, Hsinchu 300, Taiwan.
Yung-Liang
YL
Chen
School of Pharmacy, China Medical University, Taichung 404, Taiwan.
Fu-Shin
FS
Chueh
Department of Pharmacology, China Medical University, Taichung 404, Taiwan.
Jai-Sing
JS
Yang
Department of Clinical Pathology, Cheng Hsin General Hospital, Taipei 112, Taiwan.
Shu-Ching
SC
Hsueh
Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan.
Chi-Cheng
CC
Lu
Department of Life Sciences, National Chung Hsing University, Taichung 402, Taiwan.
Jo-Hua
JH
Chiang
Department of Restaurant, Hotel and Institutional Management, Fu-Jen Catholic University, New Taipei 242, Taiwan.
Ching-Sung
CS
Lee
Department of Clinical Pathology, Cheng Hsin General Hospital, Taipei 112, Taiwan.|Department of Restaurant, Hotel and Institutional Management, Fu-Jen Catholic University, New Taipei 242, Taiwan.
Hsu-Feng
HF
Lu
Department of Biological Science and Technology, China Medical University, Taichung 404, Taiwan.|Department of Biotechnology, Asia University, Taichung 413, Taiwan.
Jing-Gung
JG
Chung
D000818:Animals; D000972:Antineoplastic Agents, Phytogenic / Q000627:therapeutic use / Q000633:toxicity*; D017209:Apoptosis / Q000187:drug effects*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism; D002166:Camptothecin / Q000031:analogs & derivatives* / Q000627:therapeutic use / Q000633:toxicity; D053148:Caspase 3 / Q000378:metabolism; D018797:Cell Cycle Proteins / Q000378:metabolism; D004249:DNA Damage / Q000187:drug effects; D004195:Disease Models, Animal; D000091342:Endoplasmic Reticulum Chaperone BiP; D059585:G1 Phase Cell Cycle Checkpoints / Q000187:drug effects; D018922:HL-60 Cells; D006801:Humans; D000077146:Irinotecan; D015473:Leukemia, Promyelocytic, Acute / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology; D008297:Male; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D018613:Microscopy, Confocal; D014183:Transplantation, Heterologous
D000972:Antineoplastic Agents, Phytogenic; D051017:Apoptosis Regulatory Proteins; D018797:Cell Cycle Proteins; D000091342:Endoplasmic Reticulum Chaperone BiP; C000717247:HSPA5 protein, human; C000717248:Hspa5 protein, mouse; D000077146:Irinotecan; D053148:Caspase 3; D002166:Camptothecin
antileukemia; apoptosis; in vitro; irinotecan HCl; tumor xenograft model