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[Cancer Research 56, 5079-5086, November 1, 1996]
© 1996 American Association for Cancer Research
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A Common Mutant Epidermal Growth Factor Receptor Confers Enhanced Tumorigenicity on Human Glioblastoma Cells by Increasing Proliferation and Reducing Apoptosis1

Motoo Nagane, Frank Coufal, Hong Lin, Oliver Bögler, Webster K. Cavenee and H-J. Su Huang2

Ludwig Institute for Cancer Research [M. N., F. C., H. L., O. B., W. K. C., H-J. S. H], Department of Medicine [W. K. C., H-J. S. H.], and Center for Molecular Genetics [W. K. C.], University of California at San Diego, La Jolla, CA 92093-0660

Alterations of the EGFR gene occur frequently in human gliomas where the most common is an in-frame deletion of exons 2–7 from the extracellular domain, resulting in a truncated mutant receptor ({Delta}EGFR or de 2–7 EGFR). We previously demonstrated that introduction of {Delta}EGFR into human U87MG glioblastoma cells (U87MG.{Delta}EGFR) conferred remarkably enhanced tumorigenicity in vivo. Here, we show by cell-mixing experiments that the enhanced tumorigenicity conferred by {Delta}EGFR is attributable to a growth advantage intrinsic to cells expressing the mutant receptor. We analyzed the labeling index of the proliferation markers Ki-67 and bromodeoxyuridine and found that tumors derived from U87MG.{Delta}EGFR cells had significantly higher labeling indexes than those of tumors derived from U87MG cells that were either naive, expressed kinase-deficient mutants of {Delta}EGFR, or overexpressed exogenous wild-type EGFR. We also utilized terminal deoxynucleotidyl transferase-mediated nick end-labeling assays and showed that the apoptotic index of U87MG.{Delta}EGFR tumors was more than 4-fold lower than that of parental U87MG tumors. This decrease in cell death was inversely correlated with the expression level of Bcl-XL, a negative regulator of apoptosis, which was more than 3-fold higher in U87MG.{Delta}EGFR-derived tumors than in those derived from parental cells. Similar observations were obtained in vitro in serum-free conditions. These results suggest that {Delta}EGFR exerts its pronounced enhancement of glioblastoma tumorigenicity by stimulating proliferation and inhibiting apoptosis and that the effects are directly attributable to its constitutively active signal.

1 This work was supported in part by the Yasuda Medical Research Foundation, Osaka (to M. N.), NIH Training Grant 5-T32-NS 07342 (to F. C.), and California Division of the American Cancer Society Fellowship 1-62-95 (to O. B.).

2 To whom requests for reprints should be addressed, at Ludwig Institute for Cancer Research, 9500 Gilman Drive, La Jolla, CA 92093-0660.

Received 5/13/96. Accepted 8/30/96.




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[Abstract] [Full Text] [PDF]


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