title: Knockdown of Bcl-xL enhances growth-inhibiting and apoptosis-inducing effects of resveratrol and clofarabine in malignant mesothelioma H-2452 cells.
abstract: Mcl-1 and Bcl-xL, key anti-apoptotic proteins of the Bcl-2 family, have attracted attention as important molecules in the cell survival and drug resistance. In this study, we investigated whether inhibition of Bcl-xL influences cell growth and apoptosis against simultaneous treatment of resveratrol and clofarabine in the human malignant mesothelioma H-2452 cells. Resveratrol and clofarabine decreased Mcl-1 protein levels but had little effect on Bcl-xL levels. In the presence of two compounds, any detectable change in the Mcl-1 mRNA levels was not observed in RT-PCR analysis, whereas pretreatment with the proteasome inhibitor MG132 led to its accumulation to levels far above basal levels. The knockdown of Bcl-xL inhibited cell proliferation with cell accumulation at G2/M phase and the appearance of sub-G0/G1 peak in DNA flow cytometric assay. The suppression of cell growth was accompanied by an increase in the caspase-3/7 activity with the resultant cleavages of procaspase-3 and its substrate poly (ADP-ribose) polymerase, and increased percentage of apoptotic propensities in annexin V binding assay. Collectively, our data represent that the efficacy of resveratrol and clofarabine for apoptosis induction was substantially enhanced by Bcl-xL-lowering strategy in which the simultaneous targeting of Mcl-1 and Bcl-xL could be a more effective strategy for treating malignant mesothelioma.
authors.affiliation: Soonchunhyung Environmental Health Center for Asbestos, Soonchunhyang University Cheonan Hospital, Cheonan, Korea. ; Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Cheonan, Korea.
authors.forename: Yoon-Jin
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheonan, Korea.
authors.forename: In-Sung
authors.identifier: 0000-0002-1234-1828
authors.initials: IS
authors.lastname: Hwang
authors.affiliation: Soonchunhyung Environmental Health Center for Asbestos, Soonchunhyang University Cheonan Hospital, Cheonan, Korea.
authors.forename: Yong-Jin
authors.identifier: 0000-0002-0911-2485
authors.initials: YJ
authors.lastname: Lee
authors.affiliation: Department of Urology, Soonchunhyang University Cheonan Hospital, Cheonan, Korea.
authors.forename: Chang-Ho
authors.identifier: 0000-0003-3189-5532
authors.initials: CH
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Soonchunhyang University, Asan, Korea.
authors.forename: Sung-Ho
authors.identifier: 0000-0001-8989-6533
authors.initials: SH
authors.lastname: Kim
authors.affiliation: Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Cheonan, Korea.
authors.forename: Hae-Saeon
authors.identifier: 0000-0002-2498-2147
authors.initials: HS
authors.lastname: Nam
authors.affiliation: Division of Molecular Cancer Research, Soonchunhyang Medical Research Institute, Cheonan, Korea.
authors.forename: Young-Jin
authors.identifier: 0000-0002-7076-5725
authors.initials: YJ
authors.lastname: Choi
authors.affiliation: Soonchunhyung Environmental Health Center for Asbestos, Soonchunhyang University Cheonan Hospital, Cheonan, Korea. ; Department of Biochemistry, College of Medicine, Soonchunhyang University, Cheonan, Korea.
authors.forename: Sang-Han
authors.identifier: 0000-0001-6407-9959
authors.initials: SH
authors.lastname: Lee
mesh_terms: D000227:Adenine Nucleotides / Q000494:pharmacology*; D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D001087:Arabinonucleosides / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D000077866:Clofarabine; D059565:G2 Phase Cell Cycle Checkpoints / Q000187:drug effects; D055785:Gene Knockdown Techniques; D006801:Humans; D007976:Leupeptins / Q000494:pharmacology; D008175:Lung Neoplasms / Q000378:metabolism / Q000473:pathology; D059566:M Phase Cell Cycle Checkpoints / Q000187:drug effects; D008654:Mesothelioma / Q000378:metabolism / Q000473:pathology; D000086002:Mesothelioma, Malignant; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D000077185:Resveratrol; D013267:Stilbenes / Q000494:pharmacology*; D051020:bcl-X Protein / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism*
chemical_list: D000227:Adenine Nucleotides; D000964:Antimetabolites, Antineoplastic; D001087:Arabinonucleosides; D007976:Leupeptins; C579095:MCL1 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D013267:Stilbenes; D051020:bcl-X Protein; D000077866:Clofarabine; D053148:Caspase 3; D053179:Caspase 7; D000077185:Resveratrol; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde
keywords: Apoptosis; Bcl-xL; Clofarabine; Mcl-1; Mesothelioma, Malignant; Resveratol