Enhancement of chemosensitivity toward peplomycin by calpastatin-stabilized NF-kappaB p65 in esophageal carcinoma cells: possible involvement of Fas/Fas-L synergism.
Chemosensitivity to anticancer drugs was compared between two human esophageal carcinoma cell lines, T.Tn and YES-6 cells. T.Tn cells were more resistant than YES-6 cells to peplomycin (PEP) but not to the other anticancer drugs such as camptothecin, mitomycin C and cytosine arabinoside. Western blot analysis showed higher expression levels of m-calpain and activated mu-calpain in T.Tn cells than in YES-6 cells. On the other hand, YES-6 cells showed a high expression level of calpastatin, which is a calpain-specific endogenous inhibitor. To investigate whether calpain activity was involved in the chemosensitivity, T.Tn cells were transfected with calpastatin cDNA in an inducible expression vector. The induction of calpastatin was accompanied by increased chemosensitivity to PEP. The increases in calpastatin levels were followed by serial increases in the expression levels of NF-kappaB p65 and Fas. Since purified m- or mu-calpain degraded NF-kappaB p65 in vitro, it is possible that calpastatin suppressed calpain-mediated degradation of NF-kappaB p65. Fas ligand (Fas-L) protein levels increased after treatment of the parental T.Tn and calpastatin-transfected cells with PEP, suggesting the synergism between calpastatin-induced Fas and PEP-induced Fas-L. These results suggest that calpain/calpastatin expression levels are effective markers for predicting the sensitivity of human esophageal carcinoma cells to PEP.
Department of Biochemistry and Genetics, Graduate School of Medicine, Chiba University, Chiba, 260-8670, Japan.
T-L
TL
Liu
H
H
Shimada
T
T
Ochiai
T
T
Shiratori
S-E
SE
Lin
M
M
Kitagawa
K
K
Harigaya
M
M
Maki
M
M
Oka
T
T
Abe
M
M
Takiguchi
T
T
Hiwasa
D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D002135:Calcium-Binding Proteins / Q000494:pharmacology*; D002277:Carcinoma / Q000188:drug therapy; D045744:Cell Line, Tumor; D004938:Esophageal Neoplasms / Q000188:drug therapy*; D053222:Fas Ligand Protein / Q000378:metabolism*; D006801:Humans; D017663:Peplomycin / Q000494:pharmacology*; D051996:Transcription Factor RelA / Q000502:physiology*; D019014:fas Receptor / Q000378:metabolism*
D000903:Antibiotics, Antineoplastic; D002135:Calcium-Binding Proteins; C491901:FAS protein, human; D053222:Fas Ligand Protein; D051996:Transcription Factor RelA; D019014:fas Receptor; D017663:Peplomycin; C033668:calpastatin