Inhibition of adenovirus-mediated p27kip1 gene on growth of esophageal carcinoma cell strain

World J Gastroenterol. 2003 Nov;9(11):2404-8. doi: 10.3748/wjg.v9.i11.2404.

Abstract

Aim: 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.

Conclusion: 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.

MeSH terms

  • Adenoviridae / genetics
  • Apoptosis
  • Cell Cycle Proteins / genetics*
  • Cell Division / physiology
  • Cell Line, Tumor / cytology
  • Cell Line, Tumor / physiology
  • Cyclin-Dependent Kinase Inhibitor p27
  • Esophageal Neoplasms*
  • Gene Expression Regulation, Neoplastic
  • Humans
  • In Situ Nick-End Labeling
  • Leucine / pharmacokinetics
  • Plasmids
  • Polymerase Chain Reaction
  • Thymidine / pharmacokinetics
  • Transfection
  • Tritium
  • Tumor Suppressor Proteins / genetics*

Substances

  • Cell Cycle Proteins
  • Tumor Suppressor Proteins
  • Tritium
  • Cyclin-Dependent Kinase Inhibitor p27
  • Leucine
  • Thymidine