Effect of leptin in proliferating and differentiated HC11 mouse mammary cells.
Leptin and its receptors have been shown to be expressed in several tissues thus suggesting that this protein might be effective not only at the CNS level, but also peripherically. We demonstrated by RT-PCR analysis that leptin and its long isoform receptor are expressed in the mouse mammary epithelial cell line HC11, an in vitro cell model considered suitable to study the regulation of the functional development of the mammary epithelium. Furthermore, leptin secretion by HC11 cells was demonstrated by heterologous ELISA. Neither mRNA expression nor protein secretion changed throughout the different phases of differentiation of the cell line. Receptor mRNA was not modified when cells were induced to express beta-casein. High concentrations of leptin (between 1.5 and 15 microM) significantly (p<0.05) reduced cell growth as measured by MTT test. HC11 cells were transfected with pbetacCAT, a chimeric rat-beta casein gene promoter-CAT gene construct and CAT ELISA was used to determine gene expression. Leptin, from 1.5 nM to 15 microM, was shown to positively (p<0.05) influence beta-casein expression both in the presence or in the absence of prolactin. These data provide evidence that leptin, through its receptor, may be an important mediator in regulating mammary gland growth and development.
Baratta
Mario
M
Department of Veterinary Morphophysiology, University of Turin, Via Leonardo da Vinci 44, I-10095, Grugliasco, Italy. baratta@veter.unito.it
Grolli
Stefano
S
Tamanini
Carlo
C
D000818:Animals; D002364:Caseins / Q000235:genetics / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002455:Cell Division / Q000187:drug effects*; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004847:Epithelial Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005260:Female; D020738:Leptin / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D008321:Mammary Glands, Animal / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D011401:Promoter Regions, Genetic / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D054411:Receptors, Leptin; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013778:Tetrazolium Salts; D013844:Thiazoles
D002364:Caseins; D020738:Leptin; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D054411:Receptors, Leptin; D011993:Recombinant Fusion Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; C515447:leptin receptor, mouse; D015500:Chloramphenicol O-Acetyltransferase; C022616:thiazolyl blue