Role of MEK/ERK pathway in the MAD2-mediated cisplatin sensitivity in testicular germ cell tumour cells.
Testicular germ cell tumour (TGCT) is the most common malignancy in young males. Although most TGCTs are sensitive to cisplatin-based chemotherapy, significant numbers of TGCT patients still relapse and die each year because of the development of resistance to cisplatin. Previously, we first reported that a key regulator of the mitotic checkpoint, mitotic arrest deficient-2 (MAD2), was a mediator of cisplatin sensitivity in human cancer cells. In this study, we investigated whether MAD2 played a role in cellular sensitivity to cisplatin in TGCT cells and the underlying molecular mechanisms responsible. Using 10 TGCT cell lines, we found that increased MAD2 expression was correlated with cellular sensitivity to cisplatin, which was associated with activation of the MEK pathway. Treatment of cells expressing high levels of MAD2 with an MEK inhibitor, U0126, led to cellular protection against cisplatin-induced apoptosis. Inactivation of MAD2 by transfecting a dominant-negative construct in TGCT cells with high levels of MAD2 resulted in the suppression of MEK pathway and resistance to cisplatin-induced cell death. These results support previous suggestion on the involvement of mitotic checkpoint in DNA damage response in human cancer cells and demonstrate a possible molecular mechanism responsible for the MAD2-mediated sensitivity to cisplatin in TGCT cells. Our results also suggest that downregulation of MAD2 may be an indicator for identification of TGCT cancer cells that are potentially resistant to cisplatin-based therapy.
Fung
M K L
MK
Cancer Biology Group, Department of Anatomy, Laboratory Block, Faculty of Medicine, The University of Hong Kong, 21 Sassoon Road, Hong Kong, SAR, China.
Cheung
H-W
HW
Ling
M-T
MT
Cheung
A L M
AL
Wong
Y-C
YC
Wang
X
X
D002070:Butadienes / Q000494:pharmacology; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D018797:Cell Cycle Proteins / Q000494:pharmacology*; D045744:Cell Line, Tumor; D002945:Cisplatin / Q000627:therapeutic use*; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm; D006801:Humans; D048369:MAP Kinase Kinase 1 / Q000378:metabolism*; D064246:Mad2 Proteins; D008297:Male; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D008954:Models, Biological; D009373:Neoplasms, Germ Cell and Embryonal; D009570:Nitriles / Q000494:pharmacology; D012097:Repressor Proteins / Q000494:pharmacology*; D015398:Signal Transduction; D013736:Testicular Neoplasms / Q000188:drug therapy*; D014162:Transfection
D002070:Butadienes; D002135:Calcium-Binding Proteins; D018797:Cell Cycle Proteins; C577697:MAD2L1 protein, human; D064246:Mad2 Proteins; D009570:Nitriles; D012097:Repressor Proteins; C113580:U 0126; D048052:Mitogen-Activated Protein Kinase 3; D048369:MAP Kinase Kinase 1; D002945:Cisplatin