Mechanism of apoptotic cell death of human gastric carcinoma cells mediated by transforming growth factor beta.
Human gastric carcinoma cell line HSC-39 has been shown to undergo apoptotic cell death in response to treatment with transforming growth factor beta1 (TGF-beta1). To understand better the cell death mechanism in this TGF-beta1-mediated apoptosis, we investigated the effect of the expression of TGF-beta-stimulated clone 22 (TSC-22) on cell death events. TGF-beta1 induced TSC-22 gene expression in HSC-39 cells only when the cells had previously been adapted to the serum-free culture conditions required to undergo TGF-beta1-mediated apoptosis. HSC-39 cells transfected with a TSC-22 expression vector showed a significant decrease in cell viability compared with those transfected with a control vector. The cellular events characteristic of apoptosis, chromatin condensation and DNA fragmentation were observed only in cells transfected with a TSC-22 expression vector. On immunostaining of the transfected cells, almost every cell that expressed TSC-22 tagged with influenza virus haemagglutinin exhibited the morphology of an apoptotic cell. Partial protection from the cell death effect of TGF-beta1 on HSC-39 cells was observed when cells were treated with acetyl-L-aspartyl-L-glutamyl-L-valyl-L-aspart-1-al (Ac-DEVD-CHO, an inhibitor specific for CPP32-type protease). Protection against cell death by the transfection of a TSC-22 expression vector was also offered by Ac-DEVD-CHO addition. These results suggest that TSC-22 elicits the apoptotic cell death of human gastric carcinoma cells through the activation of CPP32-like protease and mediates the TGF-beta1 signalling pathway to apoptosis.
Ohta
S
S
Shionogi Research Laboratories, Shionogi and Co., Ltd., 5-12-4 Sagisu, Fukushima-ku, Osaka, Osaka 553, Japan.
Yanagihara
K
K
Nagata
K
K
D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases / Q000378:metabolism; D004792:Enzyme Precursors / Q000378:metabolism; D005822:Genetic Vectors; D006801:Humans; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000494:pharmacology; D012097:Repressor Proteins; D013274:Stomach Neoplasms / Q000201:enzymology / Q000473:pathology*; D014157:Transcription Factors / Q000235:genetics; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D004792:Enzyme Precursors; D012333:RNA, Messenger; D011994:Recombinant Proteins; D012097:Repressor Proteins; C099726:TSC22D1 protein, human; D014157:Transcription Factors; D016212:Transforming Growth Factor beta; C505407:CASP3 protein, human; D053148:Caspase 3; D020169:Caspases; D003546:Cysteine Endopeptidases