Category BI L06 The Effects of Epidermal Growth Factor (EGF)-like Growth Factor

Ligands on EGF

Abstract Epidermal growth factor receptor (EGFR) is often found to be over

expressed in various cancers resulting in accumulation of EGFR to the

plasma membranes of malignant epithelial tumors. Previous research has

shown that one of the seven EGFR ligands, amphiregulin (AREG),

promotes EGFR recycling to the plasma membrane over receptor

internalization and degradation as observed following “canonical” EGF

stimulation of EGFR. In this study, we used six different EGF-like ligands to

investigate their effects on modulating cell growth and EGFR localization in

SUM-149 cells expressing AREG short hairpin RNA (shRNA), or knock-

down cells, in which AREG expression is muted. By western blot analysis,

growth assays, and immunofluorescent staining assays, it was found

that high concentrations of EGF promoted internalization of membrane-

localized EGFR and stunted cell growth. On the contrary, low

concentrations of EGF allowed for membrane-localized EGFR to remain on

the cell surface and enhanced cell proliferation in the AREG knock-down

cells. These results suggest that AREG expression and EGFR membrane

accumulation are independent factors. Furthermore, the results of the

immunofluorescence assays showed that SUM-149 cells, which express

endogenous AREG, and SUM-149 AREG knock-downs had similar levels

of EGFR plasma membrane accumulation. However, a morphological

difference in the manner of invasion of the SUM-149 cells and the SUM-

149 AREG knock-down cells was observed. SUM-149 cells, expressing

endogenous AREG, migrated in sheets while the knock-down cells were

more separate and spread out while migrating. These results further

suggest that AREG expression facilitates cancer cell invasion, as indicated

by the cancer cells’ movement in sheets. If this is so, AREG could be used

as a potential biomarker, allowing for specific chemotherapeutic

strategies.

Bibliography 1) Mendelsohn, Baird, Fan, Markowitz. “Growth Factors and their

Receptors in Epithelial

Malignancies.” Cancer Biology. 2001 2) McBryan, Howlin, Napoletano,

Martin. “Amphiregulin: Role in Mammary Gland Development

and Breast Cancer.” Journal of Mammary Gland Biology and Neoplasia 13

(2008): 159-169. 3) Wilmarth, Baillo, Dzubinski, Wilson, Riese, Ethier.

“Altered EGFR Localization and

Degradation in Human Breast Cancer Cells with an Amphiregulin/EGFR

Autocrine Loop.” Cell

Signal 21 (2009): 212-219.
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