Pretreatment with glucocorticoid is essential for lactogenic induction of the bovine β-casein/CAT expression in HC11 cells

Hormonal regulation of the bovine beta-casein gene expression was studied in a murine mammary epithelial HC11 cells and compared with that of the rat beta-casein gene expression. CAT expression vectors driven by their promoter sequences were transfected into HC11 cells. Stable transfectents were treated with lactogenic hormones, dexamethasone and prolactin for 2 days in confluent cultures. While the lactogenic hormones synergistically induced a strong activation of the rat beta-casein/CAT expression, neither a single or combined treatment of dexamethasone and prolactin induced the bovine beta-casein/CAT expression. To test a sequential treatment effect of lactogenic hormones on the bovine beta-casein/CAT expression, cells were first heated with either dexamethasone or prolactin for Various days and then subjected to the second treatment with both hormones for 2 days. Only dexamethasone-, but not prolactin-pretreated cells showed a strong lactogenic induction. Moreover, the fold induction of dexamethasone-pretreated cells increased gradually as a function of duration of dexamethasone pretreatment. A series of the bovine beta-casein/CAT constructs with different length of the bovine beta-casein 5' flanking region ranged from 0.3 kb to about 15 kb was analyzed in 12-days dexamethasone-pretreated cultures. CAT expression was increased even in 0.3 kb-containing construct, but prominent induction was seen in more than 1.8 kb-containing constructs. Therefore, it could be concluded that a long-term dexamethasone pretreatment is essential for lactogenic induction of the bovine beta-casein expression and the 0.3 kb proximal promoter region is important, but more distal promoter element(s) is necessary for mediating the coordinated action of lactogenic hormones to the bovine beta-casein expression.