@inbook{c3a49981969e4f188e035c53c6e8b1ea,
title = "Tools to Study the Function of the Ras-Related, Estrogen-Regulated Growth Inhibitor in Breast Cancer",
abstract = "The Ras-related, estrogen-regulated growth inhibitor (Rerg) is a Ras-related small GTPase and candidate tumor suppressor. Rerg gene expression is stimulated by the estrogen receptor α (ERα), and Rerg gene expression is absent in ER-negative breast cancers. ER-negative breast cancers are highly invasive and metastastic and are typically more advanced than their ER-positive counterparts. Like Ras, Rerg binds and hydrolyzes GTP, but unlike Ras, Rerg has been shown to possess growth inhibitory activity in breast cancer cells. The precise role that Rerg loss plays in breast cancer growth and the mechanisms by which it does so are unknown. This chapter describes tools used to detect and manipulate the expression of Rerg in breast cancer cells. We validate use of an antibody to detect Rerg expression. We describe the generation of expression vectors that encode wild-type and mutants of Rerg that are altered in GDP/GTP regulation. We also describe the development of an inducible Rerg expression system and of a retrovirus-based RNA interference approach to repress Rerg expression. These tools will be invaluable in evaluating the biological function of Rerg in breast cancer.",
author = "Hanker, {Ariella B.} and Staeci Morita and Repasky, {Gretchen A.} and Ross, {Douglas T.} and Seitz, {Robert S.} and Der, {Channing J.}",
note = "Funding Information: We thank Douglas Andres (University of Kentucky) for the pKH3 Rerg wild type, Rerg Q64L, and Rerg S20N plasmids; Christopher Counter (Duke University) for pBabe ER H‐Ras G12V; Kevin Healy (UNC‐Chapel Hill) for the pBabe ER empty vector; Natalia Mitin (UNC‐Chapel Hill) for the pSUPERIOR.retro.puro GFP shRNA plasmid; Stephen Ethier (University of Michigan) and Carolyn Sartor (UNC‐Chapel Hill) for the SUM149 cells; and Brian Z. Ring (Applied Genomics Inc.) for assistance in manuscript writing. This work was supported in part by a Susan G. Komen Foundation grant to C.J.D. (BCTR0402537). A.B.H. is a recipient of the UNC Cancer Cell Biology Training Grant and Department of Defense Breast Cancer Research Program Predoctoral Fellowship (BC061107).",
year = "2008",
doi = "10.1016/S0076-6879(07)00405-3",
language = "English (US)",
isbn = "9780123743114",
series = "Methods in Enzymology",
pages = "53--72",
editor = "William Balch and Channing Der and Alan Hall",
booktitle = "Small GTPases in Disease, Part B",
}