Facilitation of adenoviral wild-type p53-induced apoptotic cell death by overexpression of p33(ING1) in T.Tn human esophageal carcinoma cells.
To investigate the effect of p33(ING1) on wild-type p53 gene therapy, T.Tn human esophageal carcinoma cells were stably transfected with p33(ING1) cDNA. Infection with Ad-p53 (recombinant adenovirus containing wild-type p53) into p33-transfected cells reduced cell viability, while infection with empty vector had little effect. This reduced viability was shown to be due to apoptotic cell death by the TUNEL (terminal deoxynucleotidyl transferase-mediated nick end-labeling) assay. Following infection with Ad-p53, levels of p53 were similar in p33-expressing cells and in the parental line. However, levels of p21 and Mdm2 were elevated in p33-transfected cells. Nonetheless, this enhanced expression of Mdm2 appeared to be ineffective in downregulating p53. Transient transfection with mutant Mdm2 prior to Ad-p53 infection provided a significant protection as compared with cells transfected with wild-type Mdm2. These results imply a synergistic effect between p33 and p53 in the induction of apoptosis of human esophageal carcinoma cells. A role for Mdm2 in this synergism is suggested.
Shimada
Hideaki
H
Department of Academic Surgery, Chiba University, Graduate School of Medicine, Inohana 1-8-1, Chuo-ku, Chiba 260-8670, Japan.
Liu
Tian-Ling
TL
Ochiai
Takenori
T
Shimizu
Takanori
T
Haupt
Ygal
Y
Hamada
Hirofumi
H
Abe
Toshihiro
T
Oka
Masaaki
M
Takiguchi
Masaki
M
Hiwasa
Takaki
T
D000256:Adenoviridae / Q000235:genetics*; D017209:Apoptosis; D015153:Blotting, Western; D002448:Cell Adhesion; D018797:Cell Cycle Proteins; D002455:Cell Division; D002470:Cell Survival; D003260:Contact Inhibition; D004268:DNA-Binding Proteins; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000473:pathology*; D016147:Genes, Tumor Suppressor; D016158:Genes, p53 / Q000235:genetics; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D020287:In Situ Nick-End Labeling; D000076023:Inhibitor of Growth Protein 1; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000378:metabolism; D051736:Proto-Oncogene Proteins c-mdm2; D014162:Transfection; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000378:metabolism*; D025521:Tumor Suppressor Proteins
D018797:Cell Cycle Proteins; D004268:DNA-Binding Proteins; C501503:ING1 protein, human; D000076023:Inhibitor of Growth Protein 1; D047908:Intracellular Signaling Peptides and Proteins; D009687:Nuclear Proteins; D011506:Proteins; D011518:Proto-Oncogene Proteins; D016159:Tumor Suppressor Protein p53; D025521:Tumor Suppressor Proteins; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2