Korean J Hematol 2003; 38(2):
Published online June 30, 2003
© The Korean Society of Hematology
급성백혈병 세포주에서 저용량 Doxorubicin에 의한 TRAIL 저항성의 극복
정희정, 김숙자, 김현정, 김성한, 변재호, 이남수, 김찬규, 이규택, 박성규, 원종호, 홍대식, 박희숙
순천향대학교 의과대학 임상분자생물학연구소,
순천향대학교 의과대학 종양혈액내과,
가톨릭대학교 의과대학 종양내과
Restoration of the TRAIL Resistance with a Low Dose Doxorubicin in Acute Leukemia Cell Lines
BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been reported to induce apoptosis in various tumor cells but not in normal cells, which suggests its potential use as a tumor-specific antineoplastic agent. In the present study, we examined here that the treatment of TRAIL-resistant leukemia cells with a low dose doxorubicin in combination of TRAIL induces a synergistic apoptotic response.
METHODS : Cell growth inhibition was assessed by MTT assay, and the induction of apoptosis was examined using flow cytometry after stained with Annexin V. To find out the molecular change of the TRAIL receptors and caspase expression in acute leukemia cell lines and human umbilical cord blood (UCB) mononuclear cells, RT-PCR for TRAIL receptors and western blot analysis for DR4, DR5, and caspase 3, 7, 8, and 9 were performed.
RESULTS : The Jurkat cell line was TRAIL sensitive and TRAIL-resistant Molt-4 cell line became sensitive after treatment with TRAIL and a low dose of doxorubicin (0.1 μM), but UCB mononuclear cells remained resistant. DR4 expression was increased when TRAIL-sensitive Jurkat cells were treated with TRAIL. DR5 expression was increased after exposing TRAIL- resistant Molt-4 cell line to TRAIL plus a low dose of doxorubicin for 24 hours. The expression of DR4 and DR5 in UCB mononuclear cells was unchanged after treatment with TRAIL, a low dose doxorubicin, or TRAIL plus a low dose of doxorubicin. Activated caspase 3 expression was augmented by
TRAIL plus a low dose of doxorubicin than TRAIL or a low dose of doxorubicin alone in TRAIL-resistant Molt-4 cells.
CONCLUSION : A low dose doxorubicin in combination with TRAIL may effectively promote caspase activation in TRAIL-resistant leukemia cells. Apoptosis synergistically induced by the combination of a low dose doxorubicin and TRAIL might be considered as an effective and safe tool in treating TRAIL-resistant acute leukemia.
Keywords TRAIL; Doxorubicin; Apoptosis; Acute leukemia;
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Korean J Hematol 2003; 38(2): 108-118
Published online June 30, 2003
Copyright © The Korean Society of Hematology.
급성백혈병 세포주에서 저용량 Doxorubicin에 의한 TRAIL 저항성의 극복
정희정, 김숙자, 김현정, 김성한, 변재호, 이남수, 김찬규, 이규택, 박성규, 원종호, 홍대식, 박희숙
순천향대학교 의과대학 임상분자생물학연구소,
순천향대학교 의과대학 종양혈액내과,
가톨릭대학교 의과대학 종양내과
Restoration of the TRAIL Resistance with a Low Dose Doxorubicin in Acute Leukemia Cell Lines
Hee Jeong Cheong, Sook Ja Kim, Hyun Jung Kim, Sung Han Kim, Jae Ho Byun, Nam Su Lee, Chan Kyu Kim, Kyu Taeg Lee, Sung Kyu Park, Jong Ho Won, Dae Sik Hong, Hee Sook Park
Institute for Clinical Molecular Biology Research, Hematology, Oncology, Soon Chun Hyang University College of Medicine, Seoul, Korea
Division of Oncology, The Catholic University of Korea, Seoul, Korea
Abstract
BACKGROUND: Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) has been reported to induce apoptosis in various tumor cells but not in normal cells, which suggests its potential use as a tumor-specific antineoplastic agent. In the present study, we examined here that the treatment of TRAIL-resistant leukemia cells with a low dose doxorubicin in combination of TRAIL induces a synergistic apoptotic response.
METHODS : Cell growth inhibition was assessed by MTT assay, and the induction of apoptosis was examined using flow cytometry after stained with Annexin V. To find out the molecular change of the TRAIL receptors and caspase expression in acute leukemia cell lines and human umbilical cord blood (UCB) mononuclear cells, RT-PCR for TRAIL receptors and western blot analysis for DR4, DR5, and caspase 3, 7, 8, and 9 were performed.
RESULTS : The Jurkat cell line was TRAIL sensitive and TRAIL-resistant Molt-4 cell line became sensitive after treatment with TRAIL and a low dose of doxorubicin (0.1 μM), but UCB mononuclear cells remained resistant. DR4 expression was increased when TRAIL-sensitive Jurkat cells were treated with TRAIL. DR5 expression was increased after exposing TRAIL- resistant Molt-4 cell line to TRAIL plus a low dose of doxorubicin for 24 hours. The expression of DR4 and DR5 in UCB mononuclear cells was unchanged after treatment with TRAIL, a low dose doxorubicin, or TRAIL plus a low dose of doxorubicin. Activated caspase 3 expression was augmented by
TRAIL plus a low dose of doxorubicin than TRAIL or a low dose of doxorubicin alone in TRAIL-resistant Molt-4 cells.
CONCLUSION : A low dose doxorubicin in combination with TRAIL may effectively promote caspase activation in TRAIL-resistant leukemia cells. Apoptosis synergistically induced by the combination of a low dose doxorubicin and TRAIL might be considered as an effective and safe tool in treating TRAIL-resistant acute leukemia.
Keywords: TRAIL, Doxorubicin, Apoptosis, Acute leukemia,
