title: Relaxin-like factor (RLF) is differentially expressed in the normal and neoplastic human mammary gland.
abstract: BACKGROUND
  Expression of relaxin-like factor (RLF), a member of the relaxin family, was studied in normal, benign, and malignant neoplastic human breast tissue.
  
  
  METHODS
  Reverse transcription polymerase chain reaction (RT-PCR) and nonradioactive in situ hybridization were employed to detect RLF transcripts. RLF epitopes were detected with a rabbit polyclonal antiserum generated against the putative receptor binding domain of human RLF. The RLF antiserum was characterized by Western blot analysis on human testicular and placental tissues, recombinant glutathione S-transferase-RLF fusion protein, and baculovirus-derived recombinant marmoset-RLF and marmoset-relaxin.
  
  
  RESULTS
  RT-PCR analysis revealed RLF amplicons in a cDNA library of normal human breast tissue and in malignant neoplastic breast tissue. RLF hybridization signals were localized exclusively in the tubuloalveolar and ductal breast epithelium but were absent in stromal cells. Benign breast disease displayed weaker RLF hybridization signals compared with normal tubuloalveolar breast tissue. Malignant transformation of breast epithelial tissues resulted in down-regulation of RLF gene expression. The weakest expression of RLF mRNA was observed in lymph node metastases of corresponding primary ductal carcinomas. Immunoreactive RLF was exclusively expressed in breast epithelial cells. Despite strong RLF hybridization signals, the tubuloalveolar epithelial cells of normal breast tissue displayed only very weak immunoreactive RLF. Benign breast disease showed clearly detectable levels of both RLF mRNA and immunoreactive protein. In contrast, epithelial cells in breast carcinoma and lymph node metastases displayed strong expression of immunoreactive RLF, although expression of RLF transcripts was weak.
  
  
  CONCLUSIONS
  Results demonstrated that transcriptional and posttranscriptional mechanisms affected human RLF gene expression in normal and neoplastic epithelial breast cells.
authors.affiliation: Department of Anatomy and Cell Biology, Martin Luther University Faculty of Medicine, Halle (Saale), Germany. sabine.hombach-klonisch@medizin.uni-halle.de
authors.forename: S
authors.initials: S
authors.lastname: Hombach-Klonisch
authors.forename: J
authors.initials: J
authors.lastname: Buchmann
authors.forename: S
authors.initials: S
authors.lastname: Sarun
authors.forename: B
authors.initials: B
authors.lastname: Fischer
authors.forename: T
authors.initials: T
authors.lastname: Klonisch
mesh_terms: D000818:Animals; D001665:Binding Sites; D015153:Blotting, Western; D001940:Breast / Q000378:metabolism* / Q000502:physiology; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D018270:Carcinoma, Ductal, Breast / Q000235:genetics / Q000378:metabolism / Q000556:secondary; D000939:Epitopes / Q000032:analysis; D005260:Female; D015870:Gene Expression; D006801:Humans; D017403:In Situ Hybridization; D007328:Insulin; D008198:Lymph Nodes / Q000378:metabolism; D008207:Lymphatic Metastasis; D008297:Male; D014176:Protein Biosynthesis; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000235:genetics / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011817:Rabbits; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D000939:Epitopes; D007328:Insulin; C084797:Leydig insulin-like protein; D011506:Proteins; D012333:RNA, Messenger