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M414238200v1
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Papers In Press, published online ahead of print May 11, 2005
J. Biol. Chem, 10.1074/jbc.M414238200
Submitted on December 17, 2004
Revised on April 25, 2005
Accepted on May 11, 2005

Reorganization of ErbB family and cell survival signaling following knockdown of ErbB2 in colon cancer cells

Yi Peter Hu, Srinivas Venkateswarlu, Natalia Sergina, Gillian Howell, Patricia St.Clair, Lisa E. Humphrey, Wenhui Li, Jennie Hauser, Elizabeth Zborowska, James K. V. Willson, and Michael G. Brattain

Department of Pharmacology and Therapeutics, Roswell Park Cancer Institute, Buffalo, NY 14263

Corresponding Author: michael.brattain{at}roswellpark.org

The role of ErbB family in supporting the malignant phenotype was characterized by stable transfection of a single chain antibody (ScFv5R) against ErbB2 containing a KDEL endoplasmic reticulum (ER) retention sequence into GEO human colon carcinoma cells. The antibody traps ErbB2 in the ER, thereby down-regulating cell surface ErbB2. The transfected cells showed inactivation of ErbB2 tyrosine phosphorylation and reduced heterodimerization of ErbB2 and ErbB3. This resulted in greater sensitivity to apoptosis induced by growth deprivation and delayed tumorigenicity in vivo. Furthermore, decreased heterodimerization of ErbB2 and ErbB3 led to a reorganization in ErbB function in transfected cells as heterodimerization between EGFR and ErbB3 increased while ErbB3 activation remained almost the same. Importantly, elimination of ErbB2 signaling resulted in an increase in EGFR expression and activation in transfected cells. Increased EGFR activation contributed to the sustained cell survival in transfected cells.


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