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CLOCK regulates mammary epithelial cell growth and differentiation
Year:
2016
Authors :
Mabjeesh, Sameer Jermaya
;
.
Shamay, Avi
;
.
Volume :
311
Co-Authors:
Casey, T., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Crodian, J., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Trujillo, A.S., Department of Animal Science, Universidad de Las Palmas de Gran Canaria, Arucas, Canary Islands, Spain
Erickson, E., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Weldon, B., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Crow, K., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Cummings, S., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Chen, Y., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Shamay, A., Department of Ruminant, Agriculture Research Organization, Volcani Center, Bet Dagan, Israel
Mabjeesh, S.J., Department of Ruminant, Agriculture Research Organization, Volcani Center, Bet Dagan, Israel, Department of Animal Sciences, The Robert H. Smith Faculty of Agriculture, Food, and Environment, The Hebrew University of Jerusalem, Rehovot, Israel
Plaut, K., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Facilitators :
From page:
To page:
(
Total pages:
1
)
Abstract:
Circadian clocks influence virtually all physiological processes, including lactation. Here, we investigate the role of the CLOCK gene in regulation of mammary epithelial cell growth and differentiation. Comparison of mammary morphology in late-pregnant wild-type and ClockΔ19 mice, showed that gland development was negatively impacted by genetic loss of a functional timing system. To understand whether these effects were due, in part, to loss of CLOCK function in the gland, the mouse mammary epithelial cell line, HC11, was transfected with short hairpin RNA that targeted Clock (shClock). Cells transfected with shClock expressed 70% less Clock mRNA than wild-type (WT) HC11 cultures, which resulted in significantly depressed levels of CLOCK protein (P < 0.05). HC11 lines carrying shClock had four-fold higher growth rates (P < 0.05), and the percentage of cells in G1 phase was significantly higher (90.1 ± 1.1% of shClock vs. 71.3 ± 3.6% of WT-HC11) following serum starvation. Quantitative-PCR (qPCR) analysis showed shClock had significant effects (P < 0.0001) on relative expression levels of Ccnd1, Wee1, and Tp63. qPCR analysis of the effect of shClock on Fasn and Cdh1 expression in undifferentiated cultures and cultures treated 96 h with dexamethasone, insulin, and prolactin (differentiated) found levels were reduced by twofold and threefold, respectively (P < 0.05), in shClock line relative to WT cultures. Abundance of CDH1 and TP63 proteins were significantly reduced in cultures transfected with shClock. These data support how CLOCK plays a role in regulation of epithelial cell growth and differentiation in the mammary gland. © 2016 the American Physiological Society.
Note:
Related Files :
Circadian
CLOCK
lactation
Mammary development
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More details
DOI :
10.1152/ajpregu.00032.2016
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
24140
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:05
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Scientific Publication
CLOCK regulates mammary epithelial cell growth and differentiation
311
Casey, T., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Crodian, J., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Trujillo, A.S., Department of Animal Science, Universidad de Las Palmas de Gran Canaria, Arucas, Canary Islands, Spain
Erickson, E., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Weldon, B., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Crow, K., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Cummings, S., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Chen, Y., Department of Animal Science, Purdue University, West Lafayette, IN, United States
Shamay, A., Department of Ruminant, Agriculture Research Organization, Volcani Center, Bet Dagan, Israel
Mabjeesh, S.J., Department of Ruminant, Agriculture Research Organization, Volcani Center, Bet Dagan, Israel, Department of Animal Sciences, The Robert H. Smith Faculty of Agriculture, Food, and Environment, The Hebrew University of Jerusalem, Rehovot, Israel
Plaut, K., Department of Animal Science, Purdue University, West Lafayette, IN, United States
CLOCK regulates mammary epithelial cell growth and differentiation
Circadian clocks influence virtually all physiological processes, including lactation. Here, we investigate the role of the CLOCK gene in regulation of mammary epithelial cell growth and differentiation. Comparison of mammary morphology in late-pregnant wild-type and ClockΔ19 mice, showed that gland development was negatively impacted by genetic loss of a functional timing system. To understand whether these effects were due, in part, to loss of CLOCK function in the gland, the mouse mammary epithelial cell line, HC11, was transfected with short hairpin RNA that targeted Clock (shClock). Cells transfected with shClock expressed 70% less Clock mRNA than wild-type (WT) HC11 cultures, which resulted in significantly depressed levels of CLOCK protein (P < 0.05). HC11 lines carrying shClock had four-fold higher growth rates (P < 0.05), and the percentage of cells in G1 phase was significantly higher (90.1 ± 1.1% of shClock vs. 71.3 ± 3.6% of WT-HC11) following serum starvation. Quantitative-PCR (qPCR) analysis showed shClock had significant effects (P < 0.0001) on relative expression levels of Ccnd1, Wee1, and Tp63. qPCR analysis of the effect of shClock on Fasn and Cdh1 expression in undifferentiated cultures and cultures treated 96 h with dexamethasone, insulin, and prolactin (differentiated) found levels were reduced by twofold and threefold, respectively (P < 0.05), in shClock line relative to WT cultures. Abundance of CDH1 and TP63 proteins were significantly reduced in cultures transfected with shClock. These data support how CLOCK plays a role in regulation of epithelial cell growth and differentiation in the mammary gland. © 2016 the American Physiological Society.
Scientific Publication
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