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© 1998 Oxford University Press

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Bcl-2 and Bcl-xL in Peroxide-Resistant A549 and U87MG Cells

Heidi K. Bojes*, P. K. Suresh*, Edward M. Mills{dagger}, Douglas R. Spitz{ddagger}, Julia E. Sim{ddagger} and James P. Kehrer*

*Division of Pharmacology and Toxicology, College of Pharmacy, The University of Texas at Austin Austin, Texas 78712-1074 {dagger}Department of Medicinal Chemistry and Molecular Pharmacology, School of Pharmacy and Pharmacal Sciences, Purdue University West Lafayette, Indiana 47907 {ddagger}Section of Cancer Biology, Radiation Oncology Center, Washington University School of Medicine St. Louis, Missouri 63108

Received October 3, 1997; accepted December 12, 1997

Overexpression of the bcl-2 and the related bcl-xL protoonco-gene proteins enhance cell survival by inhibiting apoptosis induced by many agents including oxidants. Whether these proteins contribute to survival in oxidant-resistant cells is not known. The current study assessed the expression of bcl-2 and bcl-xL proteins in human glioblastoma U87MG cells and human lung adenocar-cinoma A549 cells selected for resistance to 0, 50, 100, 200, and 400 µM H2O2 by exposure to this oxidant one time each passage for 9 months. When examined 7 to 32 days after cessation of peroxide exposure (times when peroxide resistance was maintained), bcl-2 protein levels were significantly increased in most peroxide-resistant U87MG cells. However, the increase was not dose dependent and was not accompanied by an increase in mRNA levels. A549 cells did not express significant levels of bcl-2 protein, although bcl-2 mRNA was detectable. A549 cells expressed large amounts of bcl-xL and immunohistochemistry demonstrated extensive localization of this protein around the nucleus. However, expression of this protein was not altered in peroxide-resistant lines nor was bcl-2 protein increased to a measurable level. U87MG cells also expressed bcl-xL but it was not altered in peroxide-resistant cells. Although the increased bcl-2 protein in peroxide-resistant U87MG cells may contribute to their oxidant tolerance, the lack of a dose-response relationship, the failure to induce bcl-xL protein, and the absence of any bcl-2 or bcl-xL protein induction in peroxide-resistant A549 cells suggest these genes are not primary factors in oxidant resistance.

Key Words: bcl-2; bcl-xL; hydrogen peroxide; oxidant toxicity; A549 cells; U87MG cells.


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