Estrogen-dependent expression of the chicken very low density apolipoprotein II gene in serum-free cultures of LMH cells.
The estrogen-responsive Leghorn strain M chicken hepatoma (LMH) cell line provides a model system for studying the estrogen-dependent, liver-specific expression of avian genes. Serum-free culture conditions have been established that allow expression of apolipoprotein B, very low density apolipoprotein II (apoVLDLII), serum albumin, and transferrin at levels detectable by Northern blot analysis. Regulation of apoVLDLII mRNA by estrogen occurred in an appropriate time- and dose-dependent manner in serum-free cultures of the LMH cells. The expression of apoVLDLII mRNA in serum-free culture was at least 100-fold higher than that expressed in cultures containing 10% serum. The level of estrogen receptors in LMH cells cultured with 10% serum was approximately 2000 receptors per cell, and in serum-free culture approximately 1000 receptors per cell. When these cells were transfected with estrogen receptor DNA and cultured in serum-free medium, apoVLDLII mRNA was decreased relative to that expressed in cells transfected with a control plasmid. These results indicate that when the LMH cells are cultured without serum, estrogen receptors are not the limiting factor for the expression of the apoVLDLII gene.
Berkowitz
E A
EA
Department of Biochemistry, School of Medicine, West Virginia University, Morgantown 26506.
Evans
M I
MI
D000818:Animals; D016633:Apolipoprotein A-II / Q000235:genetics*; D015152:Blotting, Northern; D002645:Chickens; D016895:Culture Media, Serum-Free; D004305:Dose-Response Relationship, Drug; D004967:Estrogens / Q000494:pharmacology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D008114:Liver Neoplasms, Experimental / Q000235:genetics* / Q000473:pathology* / Q000648:ultrastructure; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D011960:Receptors, Estrogen / Q000032:analysis; D012709:Serum Albumin / Q000235:genetics; D014168:Transferrin / Q000235:genetics; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000473:pathology / Q000648:ultrastructure
D016633:Apolipoprotein A-II; D016895:Culture Media, Serum-Free; D004967:Estrogens; D012333:RNA, Messenger; D011960:Receptors, Estrogen; D012709:Serum Albumin; D014168:Transferrin