Inhibition of adenovirus-mediated p27kip1 gene on growth of esophageal carcinoma cell strain.
OBJECTIVE
To investigate the inhibition of p27kip1 gene on the growth of esophageal carcinoma cell strain (EC9706).


METHODS
Recombinant adenovirus Ad-p27kip1 was constructed and transfected into esophageal carcinoma cell EC-9706, and its effect on p27kip1 expression, the growth of esophageal carcinoma cell, DNA replication, protein synthesis, cell multiplication and apoptosis were explored by means of cell growth count, 3H-TdR, 3H-Leucine incorporation, flow cytometry, DNA fragment analysis and TUNEL.


RESULTS
Recombinant adenovirus Ad-p27kip1 was successfully constructed with a virus titer of 1.24 X 10(12) pfu/ml. p27kip protein expression increased markedly after EC-9706 transfection, while incorporation quantity of 3H-TdR and 3H-Leucine decreased significantly. The growth of esophageal carcinoma cell was inhibited obviously. Testing of flow cytometry displayed a typical apoptosis peak, and DNA gel electrophoresis showed a typical apoptosis ladder. TUNEL showed the apoptosis rate of Ad-p27kip1 group and control group to be 37.3% and 1.26% (P<0.001) respectively.


CONCLUSIONS
Ad-p27kip1 can inhibit the growth and multiplication of esophageal carcinoma cells and induce apoptosis. Therefore, enhanced p27kip1 expression may be a new way to treat esophageal carcinoma.
Wu
Qing-Ming
QM
Department of Gastroenterology, Renmin Hospital of Wuhan University, Hubei Province, China. wuhe9224@sina.com
Yu
Jie-Ping
JP
Tong
Qiang
Q
Wang
Xiao-Hu
XH
Xie
Guo-Jian
GJ
D000256:Adenoviridae / Q000235:genetics; D017209:Apoptosis; D018797:Cell Cycle Proteins / Q000235:genetics*; D002455:Cell Division / Q000502:physiology; D045744:Cell Line, Tumor / Q000166:cytology / Q000502:physiology; D050760:Cyclin-Dependent Kinase Inhibitor p27; D004938:Esophageal Neoplasms; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D020287:In Situ Nick-End Labeling; D007930:Leucine / Q000493:pharmacokinetics; D010957:Plasmids; D016133:Polymerase Chain Reaction; D013936:Thymidine / Q000493:pharmacokinetics; D014162:Transfection; D014316:Tritium; D025521:Tumor Suppressor Proteins / Q000235:genetics*
D018797:Cell Cycle Proteins; D025521:Tumor Suppressor Proteins; D014316:Tritium; D050760:Cyclin-Dependent Kinase Inhibitor p27; D007930:Leucine; D013936:Thymidine