Abstract
Within the large group of Estrogen Receptor alpha (ERα)-negative breast cancer patients, there is a subgroup carrying the phenotype ERα(-), PR(-), and Her2(-), named accordingly "Triple-Negative" (TN). Using cell lines derived from this TN group, we wished to establish cell clones, in which ERα is ectopically expressed, forming part of a synthetic lethality screening system. Initially, we generated cell transfectants expressing a mono-cistronic ERα transcription unit, adjacent to a separate dominant selectable marker transcription unit. However, the yield of ERα expressing colonies was rather low (5-12.5%), and only about half of these displayed stable ectopic ERα expression over time. Generation and maintenance of such cell clones under minimal exposure to the ERα ligand, did not improve yield or expression stability. Indeed, other groups have also reported grave difficulties in obtaining ectopic expression of ERα in ERα-deficient breast carcinoma cells. We therefore switched to transfecting these cell lines with pERα-IRES, a plasmid vector encoding a bicistronic translation mRNA template: ERα Open Reading Frame (ORF) being upstream followed by a dominant-positive selectable marker (hygro(R)) ORF, directed for translation from...Continue Reading
References
Nov 1, 1978·In Vitro·R CailleauQ V Cruciger
Dec 1, 1992·Proceedings of the National Academy of Sciences of the United States of America·M GarciaH Rochefort
Apr 15, 1992·Journal of the National Cancer Institute·S Y Jiang, V C Jordan
Jan 1, 1990·European Journal of Cancer : Official Journal for European Organization for Research and Treatment of Cancer (EORTC) [and] European Association for Cancer Research (EACR)·M W Madsen, P Briand
Oct 6, 1995·The Journal of Biological Chemistry·H WangS A Khan
Oct 6, 1995·The Journal of Biological Chemistry·C MatsuiW K Hoeffler
Jan 1, 1996·BioTechniques·S ReesM G Lee
Dec 4, 1996·Biochemical and Biophysical Research Communications·V GurtuG Zhang
Nov 14, 1997·Science·L H HartwellS H Friend
Aug 10, 2000·Molecular Therapy : the Journal of the American Society of Gene Therapy·H MizuguchiT Hayakawa
Mar 15, 2001·The New England Journal of Medicine·D J SlamonL Norton
Jun 8, 2001·Molecular and Cellular Biology·H Y Mak, M G Parker
Nov 6, 2003·Clinical & Experimental Metastasis·Dina Chelouche LevJanet E Price
Jul 29, 2004·BMC Biotechnology·Victorine DouinBettina Couderc
May 3, 2005·The Journal of Steroid Biochemistry and Molecular Biology·J S LewisV C Jordan
Aug 20, 2005·Nature Reviews. Cancer·William G Kaelin
Sep 9, 2006·Science·Tobias SjöblomVictor E Velculescu
Mar 9, 2007·Nature·Christopher GreenmanMichael R Stratton
Oct 4, 2007·Breast Cancer Research : BCR·Bas KreikeMarc J van de Vijver
Oct 13, 2007·Science·Laura D WoodBert Vogelstein
Feb 2, 2008·Science·Jose M SilvaKenneth Chang
Feb 8, 2008·Nature Reviews. Cancer·Thomas HelledayRicky A Sharma
Jul 23, 2008·Current Opinion in Pharmacology·Christopher J Lord, Alan Ashworth
Mar 26, 2009·British Journal of Cancer·D Canaani
Jan 7, 2010·BMC Genomics·Michael BoettcherJörg D Hoheisel
Mar 8, 2011·Cell·Tingting SunThomas F Westbrook
Jun 29, 2011·Annals of Oncology : Official Journal of the European Society for Medical Oncology·A GoldhirschUNKNOWN Panel members
Citations
Jun 6, 2016·Immunology Letters·Wen Yi WangLi Xin Li
Feb 5, 2015·Placenta·G BarrientosC L Varone
Oct 6, 2017·Oncotarget·Ronni AramDan Canaani
Jun 3, 2021·International Journal of Molecular Sciences·Guus Gijsbertus Hubert van den AkkerTim Johannes Maria Welting