Growth and cell cycle regulation of mRNA levels in GH3 cells.
In the rat anterior pituitary gland, estrogen increases both prolactin (PRL) mRNA levels and stimulates the proliferation of PRL-producing cells. The temporal sequence of these events suggests that PRL gene expression may be coordinated with cell proliferation. We investigated the relationship between cell cycle progression and the accumulation of the PRL mRNA, as well as several other mRNAs, in the rat pituitary tumor GH3 cell line. Serum-deprived cells progressed from G0 to S phase in 20-24 h following serum stimulation. During this time, beta-actin mRNA levels increased 7-fold in 5 h, then returned to basal levels prior to the beginning of S phase. Histone H1 mRNA levels increased approximately 3-fold as cells entered S phase. These data are consistent with the cell cycle-dependent regulation of beta-actin and histone H1 gene expression reported for other cell types. Glucocorticoid receptor mRNA levels were barely detectable in serum-deprived cells but rapidly increased 3- to 5-fold following serum stimulation. This increase resulted in glucocorticoid receptor mRNA levels that were equivalent to those seen in cells maintained in serum-containing medium, suggesting that serum factors regulate glucocorticoid receptor gene expression. In contrast to these changes in gene expression, the levels of PRL and growth hormone (GH) mRNAs gradually increased 2-fold while the cells progressed through G1 phase. Similarly, in cells synchronized to progress through S and G2 phases following aphidicolin treatment, histone H1 gene expression showed a specific increase in S phase cells, whereas PRL and GH mRNA levels changed little with cell cycle progression. These results indicate that the levels of PRL and GH mRNAs are not regulated in a cell cycle-dependent manner. When changes in estrogen responsiveness were determined during the cell cycle, we found that estradiol treatment was capable of increasing PRL mRNA accumulation independent of cell cycle progression and cell cycle distribution in synchronized GH3 cells. These results support the hypothesis that the hormonal regulation of PRL gene expression is not significantly affected by cell growth.
Rhode
P R
PR
Department of Chemistry, California Institute of Technology, Pasadena 91125.
Gorski
J
J
D000199:Actins / Q000378:metabolism; D000818:Animals; D001769:Blood; D015152:Blotting, Northern; D002453:Cell Cycle; D015342:DNA Probes; D004958:Estradiol / Q000502:physiology; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000378:metabolism; D006657:Histones / Q000378:metabolism; D010902:Pituitary Gland / Q000166:cytology* / Q000378:metabolism; D011388:Prolactin / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D011965:Receptors, Glucocorticoid / Q000378:metabolism; D013997:Time Factors; D014407:Tumor Cells, Cultured
D000199:Actins; D015342:DNA Probes; D006657:Histones; D012333:RNA, Messenger; D011965:Receptors, Glucocorticoid; D004958:Estradiol; D011388:Prolactin; D013006:Growth Hormone