Id3-mediated enhancement of cisplatin-induced apoptosis in a sarcoma cell line MG-63.
The molecular mechanism(s) underlying the resistance to cis-diamminedichloroplatinum (CDDP)-induced growth inhibition include DNA repair, apoptosis and cell cycle progression. Inhibitor of differentiation (Id) proteins, which belong to the group of helix-loop-helix proteins, regulate cell cycle progression, differentiation and apoptosis. We examined whether CDDP exposure modulates the expression pattern of Ids and whether ectopic expression of Ids influences CDDP-induced cell death. Cell growth was assessed by WST-8 assay kit. Reactive oxygen species (ROS) was evaluated by flow cytometry using dihydroethidium. MG-63 sarcoma cells were stimulated with CDDP for various times and Id expression was assessed by reverse transcription-polymerase chain reaction. CDDP induced a considerable transient up-regulation of Id3 mRNA, but not Id2, 1-2 h after stimulation. Enforced expression of Id3 caused the MG-63 sarcoma cells to be more sensitive to CDDP-induced growth inhibition, through generation of ROS and caspase-3 activation. Together, our results suggest that CDDP-induced cell death appears to involve Id3.
Koyama
Takashi
T
Department of Immunology, Tokyo Medical University, Shinjuku-ku, Tokyo, 160-8402, Japan.
Suzuki
Hidekazu
H
Imakiire
Atsuhiro
A
Yanase
Noriko
N
Hata
Kikumi
K
Mizuguchi
Junichiro
J
D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D001859:Bone Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects / Q000502:physiology; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000494:pharmacology*; D004789:Enzyme Activation; D006801:Humans; D051796:Inhibitor of Differentiation Proteins; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*; D012516:Osteosarcoma / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species; D014162:Transfection; D015854:Up-Regulation / Q000187:drug effects
D000970:Antineoplastic Agents; D051796:Inhibitor of Differentiation Proteins; D009363:Neoplasm Proteins; D017382:Reactive Oxygen Species; C075640:ID3 protein, human; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D002945:Cisplatin