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J Thorac Cardiovasc Surg 2001;121:0061-0067
© 2001 The American Association for Thoracic Surgery

General Thoracic Surgery

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

From the Section of Thoracic Molecular Oncology, Department of Thoracic and Cardiovascular Surgery,^a and the Department of Molecular Pathology,^b The University of Texas M.D. Anderson Cancer Center, Houston, Tex.

Supported by grants from the National Institutes of Health (grant PO1 CA78778-01A1) to J.A.R., S.G.S., and T.J.M., by Specialized Program of Research Excellence in Lung Cancer Grant P50-CA70907, by gifts to the Division of Surgery and Anesthesiology from Tenneco and Exxon for the Core Laboratory Facility, by the University of Texas M. D. Anderson Cancer Center Support Core Grant CA16672, by a sponsored research agreement with Introgen Therapeutics, and by the W. M. Keck Foundation and by the tobacco settlement funds (project 8).

Received for publication May 4, 2000. Revisions requested July 25, 2000; revisions received Aug 28, 2000. Accepted for publication Sept 9, 2000. Address for reprints: Stephen G. Swisher, MD, Department of Thoracic and Cardiovascular Surgery, The University of Texas M.D. Anderson Cancer Center, 1515 Holcombe Blvd, Box 109, Houston, TX 77030 (E-mail: sswisher{at}mdanderson.org).

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.