title: Effect of anabolics on bovine granulosa-luteal cell primary cultures.
abstract: Granulosa cell tumours are observed with increased frequency among calves slaughtered in Northern Italy. The use of illegal anabolics in breeding was taken into account as a cause of this pathology. An in vitro approach was used to detect the possible alterations of cell proliferation induced by anabolics on primary cultures of bovine granulosa-luteal cells. Cultures were treated with different concentrations of substances illegally used in cattle (17beta-estradiol, clenbuterol and boldione). Cytotoxicity was determined by means of MTT test, to exclude toxic effects induced by anabolics and to determine the highest concentration to be tested. Morphological changes were evaluated by means of routine cytology, while PCNA expression was quantified in order to estimate cell proliferation. Cytotoxic effects were revealed at the highest concentrations. The only stimulating effect on cell proliferation was detected in boldione treated cultures: after 48 h treated cells, compared to controls, showed a doubled expression of PCNA. In clenbuterol and 17beta-estradiol treated cells PCNA expression was similar to controls or even decreased. As the data suggest an alteration in cell proliferation, boldione could have a role in the early stage of pathogenesis of granulosa cell tumour in cattle.
authors.affiliation: University of Turin, Dipartimento di Patologia Animale, Grugliasco (TO), Italy. paola.pregel@unito.it
authors.forename: Paola
authors.initials: P
authors.lastname: Pregel
authors.forename: Enrico
authors.initials: E
authors.lastname: Bollo
authors.forename: Francesca Tiziana
authors.initials: FT
authors.lastname: Cannizzo
authors.forename: Antonella
authors.initials: A
authors.lastname: Rampazzo
authors.forename: Simonetta
authors.initials: S
authors.lastname: Appino
authors.forename: Bartolomeo
authors.initials: B
authors.lastname: Biolatti
mesh_terms: D045930:Anabolic Agents / Q000633:toxicity*; D000730:Androstadienes / Q000633:toxicity; D000818:Animals; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D002976:Clenbuterol / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004958:Estradiol / Q000633:toxicity; D005260:Female; D006107:Granulosa Cells / Q000187:drug effects*; D008184:Luteal Cells / Q000187:drug effects*; D018809:Proliferating Cell Nuclear Antigen / Q000096:biosynthesis; D023382:Toxicity Tests, Acute
chemical_list: D045930:Anabolic Agents; D000730:Androstadienes; D018809:Proliferating Cell Nuclear Antigen; C006573:1,4-androstadiene-3,17-dione; D004958:Estradiol; D002976:Clenbuterol