Abstract
We have developed an image-based technique for signal pathway analysis, target validation, and compound screening related to mammary epithelial cell differentiation. This technique used the advantages of optical imaging and the HC11-Lux model system. The HC11-Lux cell line is a subclone of HC11 mammary epithelial cells transfected stably with a luciferase construct of the beta-casein gene promoter (p-344/-1betac-Lux). The promoter activity was imaged optically in real time following lactogenic induction. The imaging signal intensity was closely correlated with that measured using a luminometer following protein extraction (R=0.99, P<0.0001) and consistent with the messenger RNA (mRNA) level of the endogenous beta -casein gene. Using this technique, we examined the roles of JAK2/Stat5A, Raf-1/MEK/MAKP, and PI3K/Akt signal pathways with respect to differentiation. The imaging studies showed that treatment of the cells with epidermal growth factor (EGF), AG490 (JAK2-specific inhibitor), and LY294002 (PI3K-specific inhibitor) blocked lactogenic differentiation in a dose-dependent manner. PD98059 (MEK-specific inhibitor) could reverse EGF-mediated differentiation arrest. These results indicate that these pathways are essential in ce...Continue Reading
References
Feb 19, 2000·Current Opinion in Biotechnology·S A Sundberg
Feb 1, 2002·The Journal of Biological Chemistry·Jianwu XieHallgeir Rui
Jun 10, 2003·Proceedings of the National Academy of Sciences of the United States of America·Sheng DingPeter G Schultz
Oct 22, 2003·Journal of Biomolecular Screening·Zhuyin LiBruce Baron
Nov 25, 2003·The American Journal of Pathology·Liang ShanElizabeth G Snyderwine
Jun 9, 2004·Current Opinion in Chemical Biology·Paul A Clemons
Aug 31, 2004·Journal of Cellular Physiology·Maria Grazia CerritoMary Lou Cutler
Dec 21, 2004·Breast Cancer Research and Treatment·Liang ShanElizabeth G Snyderwine
Oct 4, 2005·Nature Biotechnology·Michael J EvansBenjamin F Cravatt
May 6, 2006·Biochimica Et Biophysica Acta·Ercole CavalieriThomas Sutter
May 13, 2006·Endocrinology·Suzanne E Fenton
Aug 1, 2006·Oncogene·A RaafatR Callahan
Nov 7, 2006·Analytical Biochemistry·Michael P ValleyKeith V Wood
Nov 28, 2006·Biochimica Et Biophysica Acta·James A McCubreyRichard A Franklin
Feb 27, 2007·Oncogene·R L DillonW J Muller
Oct 13, 2007·Current Opinion in Chemical Biology·Kerstin Korn, Eberhard Krausz
Jan 30, 2008·Journal of Mammary Gland Biology and Neoplasia·Kay-Uwe Wagner, Hallgeir Rui
Mar 29, 2008·Expert Opinion on Drug Metabolism & Toxicology·James J CaliJohn Shultz
Citations
Mar 20, 2012·Biochimica Et Biophysica Acta·Alain Pauloin, Eric Chanat
Sep 10, 2014·Bioscience Reports·Xinxin LiJiwen Wang
Apr 7, 2015·Journal of Dairy Science·M H KungY T Ju