title: Transfection of Sertoli cells with androgen receptor alters gene expression without androgen stimulation.
abstract: BACKGROUND
  Androgens play an important role for the development of male fertility and gained interest as growth and survival factors for certain types of cancer. Androgens act via the androgen receptor (AR/Ar), which is involved in various cell biological processes such as sex differentiation. To study the functional mechanisms of androgen action, cell culture systems and AR-transfected cell lines are needed. Transfection of AR into cell lines and subsequent gene expression analysis after androgen treatment is well established to investigate the molecular biology of target cells. However, it remains unclear how the transfection with AR itself can modulate the gene expression even without androgen stimulation. Therefore, we transfected Ar-deficient rat Sertoli cells 93RS2 by electroporation using a full length human AR.
  
  
  RESULTS
  Transfection success was confirmed by Western Blotting, immunofluorescence and RT-PCR. AR transfection-related gene expression alterations were detected with microarray-based genome-wide expression profiling of transfected and non-transfected 93RS2 cells without androgen stimulation. Microarray analysis revealed 672 differentially regulated genes with 200 up- and 472 down-regulated genes. These genes could be assigned to four major biological categories (development, hormone response, immune response and metabolism). Microarray results were confirmed by quantitative RT-PCR analysis for 22 candidate genes.
  
  
  CONCLUSIONS
  We conclude from our data, that the transfection of Ar-deficient Sertoli cells with AR has a measurable effect on gene expression even without androgen stimulation and cause Sertoli cell damage. Studies using AR-transfected cells, subsequently stimulated, should consider alterations in AR-dependent gene expression as off-target effects of the AR transfection itself.
authors.affiliation: Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University, Frankfurter Straße 98, 35392, Giessen, Germany. Daniela.Fietz@vetmed.uni-giessen.de.
authors.forename: D
authors.initials: D
authors.lastname: Fietz
authors.affiliation: Institute of Medical Microbiology, Justus Liebig University, Giessen, Germany. Melanie.Markmann@mikrobio.med.uni-giessen.de.
authors.forename: M
authors.initials: M
authors.lastname: Markmann
authors.affiliation: Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University, Frankfurter Straße 98, 35392, Giessen, Germany. de.lang@arcor.de.
authors.forename: D
authors.initials: D
authors.lastname: Lang
authors.affiliation: Department of Gynecology and Obstetrics, Justus Liebig University, Giessen, Germany. Lutz.Konrad@gyn.med.uni-giessen.de.
authors.forename: L
authors.initials: L
authors.lastname: Konrad
authors.affiliation: Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University, Frankfurter Straße 98, 35392, Giessen, Germany. Joachim.M.Geyer@vetmed.uni-giessen.de.
authors.forename: J
authors.initials: J
authors.lastname: Geyer
authors.affiliation: Department of Clinical Andrology, Centre for Reproductive Medicine and Andrology, University Clinic Münster, Münster, Germany. kliesch@ukmuenster.de.
authors.forename: S
authors.initials: S
authors.lastname: Kliesch
authors.affiliation: Institute of Medical Microbiology, Justus Liebig University, Giessen, Germany. Trinad.Chakraborty@mikrobio.med.uni-giessen.de.
authors.forename: T
authors.initials: T
authors.lastname: Chakraborty
authors.affiliation: Institute of Medical Microbiology, Justus Liebig University, Giessen, Germany. Hamid.M.Hossain@mikrobio.med.uni-giessen.de.
authors.forename: H
authors.initials: H
authors.lastname: Hossain
authors.affiliation: Institute of Veterinary Anatomy, Histology and Embryology, Justus Liebig University, Frankfurter Straße 98, 35392, Giessen, Germany. Martin.Bergmann@vetmed.uni-giessen.de.
authors.forename: M
authors.initials: M
authors.lastname: Bergmann
mesh_terms: D000728:Androgens / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D015870:Gene Expression / Q000235:genetics; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation / Q000235:genetics*; D006801:Humans; D008297:Male; D051379:Mice; D011471:Prostatic Neoplasms / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D011944:Receptors, Androgen / Q000235:genetics*; D012708:Sertoli Cells / Q000166:cytology / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
chemical_list: D000728:Androgens; D012333:RNA, Messenger; D011944:Receptors, Androgen