Adenovirus-mediated Bak gene transfer induces apoptosis in mesothelioma cell lines

J Thorac Cardiovasc Surg. 2001 Jan;121(1):61-7. doi: 10.1067/mtc.2001.111419.


Objective: Conventional treatment for mesothelioma is largely ineffective. We therefore evaluated the novel approach of adenoviral gene transfer of the proapoptotic Bcl-2 family member Bak in mesothelioma cancer cell lines, which are sensitive and resistant to adenoviral p53.

Methods: Binary adenoviral Bak (Ad/GT-Bak and Ad/GV16) and LacZ (Ad/GT-LacZ and Ad/GV16) vectors were used for transduction of the mesothelioma cell lines I-45 (p53 resistant) and REN (p53 sensitive). Protein levels were determined by Western blotting. Apoptosis was assessed by morphologic changes, caspase-3 cleavage, and fluorescence-activated cell sorter analysis of subdiploid populations. Cell viability was determined with the XTT assay. Statistical analysis was performed with analysis of variance and the Student t test.

Results: High levels of Bak gene transfer were seen after coadministration of Ad/GT-Bak and Ad/GV16 in both mesothelioma cell lines. Apoptosis was induced 24 hours after Bak but not LacZ gene transfer ([Bak: I-45, 36%; REN, 25%] vs [LacZ: I-45, 1%; REN, 3%], P <.05]) in p53-sensitive (REN) and p53-resistant (I-45) cell lines. Cellular viability was significantly decreased 48 to 72 hours after Bak gene transfer compared with control vector in both cell lines (72 hours: Bak I-45, 1.4% +/- 1.0%, and Bak REN, 4.7% +/- 1%, vs Lac-Z I-45, 83% +/- 3%, and Lac-Z REN, 100% +/- 1%; P <.05).

Conclusions: Adenovirus-mediated overexpression of the Bak gene induces apoptosis and decreased cellular viability in p53-sensitive and p53-resistant mesothelioma cells. These data suggest that the gene transfer of proapoptotic Bcl-2 family members may represent a novel gene therapy strategy to treat mesothelioma.

Publication types

  • Comparative Study
  • Evaluation Study
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenoviridae / genetics*
  • Apoptosis* / genetics
  • Blotting, Western
  • Cell Survival
  • Flow Cytometry
  • Gene Expression
  • Genetic Vectors
  • Humans
  • Membrane Proteins / genetics*
  • Membrane Proteins / metabolism
  • Mesothelioma / genetics
  • Mesothelioma / pathology*
  • Mesothelioma / virology
  • Mutation
  • Pleural Neoplasms / genetics
  • Pleural Neoplasms / pathology*
  • Pleural Neoplasms / virology
  • Transduction, Genetic / methods*
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 / genetics
  • bcl-2 Homologous Antagonist-Killer Protein


  • BAK1 protein, human
  • Membrane Proteins
  • Tumor Suppressor Protein p53
  • bcl-2 Homologous Antagonist-Killer Protein