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Epithelial cell differentiation regulated by MicroRNA-200a in mammary glands.

Nagaoka K, Zhang H, Watanabe G, Taya K - PLoS ONE (2013)

Bottom Line: Here, we found that one miRNA, miR-200a, was relatively highly expressed in epithelial cell-rich organs such as mammary glands, lung, and kidney in mice.However, knockdown of miR-200a prevented increases in ß-casein and E-cadherin mRNA expression.Protein analysis revealed that E-cadherin signal was decreased and ZEB1 (a marker of EMT) was increased following miR-200a knockdown.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Physiology, Department of Veterinary Medicine, Tokyo University of Agriculture and Technology, Tokyo, Japan. nagaokak@cc.tuat.ac.jp

ABSTRACT
Mammary gland epithelial cells undergo periodic cycles of proliferation, differentiation, and involution. Many studies have reported that miRNAs, which are small, non-coding RNAs, influence a variety of biological processes during posttranscriptional regulation. Here, we found that one miRNA, miR-200a, was relatively highly expressed in epithelial cell-rich organs such as mammary glands, lung, and kidney in mice. In mammary glands, miR-200a expression increased during mid-pregnancy through lactation; its expression was stimulated by lactogenic hormone treatment of mammary epithelial cells. Lactogenic hormone also induced the expression of milk protein ß-casein mRNA (a marker of cell differentiation) and E-cadherin mRNA (a marker of epithelial cells). However, knockdown of miR-200a prevented increases in ß-casein and E-cadherin mRNA expression. Protein analysis revealed that E-cadherin signal was decreased and ZEB1 (a marker of EMT) was increased following miR-200a knockdown. Finally, in a three-dimensional culture system modeling lumen-containing mammary ducts, miR-200a knockdown decreased the cavity formation rate and suppressed claudin-3 and par-6b expression, indicating reduced epithelial cell polarity. These observations suggest that miR-200a is important for maintaining the epithelial cell phenotype, which contributes to lactogenic hormone induction of cellular differentiation in mammary glands.

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Tissue expression of miR-200a in mice.Mice brain, heart, lung, liver, spleen, kidney, and mammary glands were collected on day 7 of lactation (n = 3 animals), and expression of miR-23b and miR-200a were analyzed by real-time PCR.
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pone-0065127-g001: Tissue expression of miR-200a in mice.Mice brain, heart, lung, liver, spleen, kidney, and mammary glands were collected on day 7 of lactation (n = 3 animals), and expression of miR-23b and miR-200a were analyzed by real-time PCR.

Mentions: To confirm the tissue expression of miR-200a and miR-23b (as a control) in mice, we conducted real-time PCR analysis using cDNAs from brain, heart, lung, liver, spleen, kidney, and mammary glands (Fig. 1). The results showed high expression of miR-200a in the lung, kidney, and mammary glands, which are organs composed of epithelial cells. miR-23b expression was observed in most tissues except in the liver and mammary glands.


Epithelial cell differentiation regulated by MicroRNA-200a in mammary glands.

Nagaoka K, Zhang H, Watanabe G, Taya K - PLoS ONE (2013)

Tissue expression of miR-200a in mice.Mice brain, heart, lung, liver, spleen, kidney, and mammary glands were collected on day 7 of lactation (n = 3 animals), and expression of miR-23b and miR-200a were analyzed by real-time PCR.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3672172&req=5

pone-0065127-g001: Tissue expression of miR-200a in mice.Mice brain, heart, lung, liver, spleen, kidney, and mammary glands were collected on day 7 of lactation (n = 3 animals), and expression of miR-23b and miR-200a were analyzed by real-time PCR.
Mentions: To confirm the tissue expression of miR-200a and miR-23b (as a control) in mice, we conducted real-time PCR analysis using cDNAs from brain, heart, lung, liver, spleen, kidney, and mammary glands (Fig. 1). The results showed high expression of miR-200a in the lung, kidney, and mammary glands, which are organs composed of epithelial cells. miR-23b expression was observed in most tissues except in the liver and mammary glands.

Bottom Line: Here, we found that one miRNA, miR-200a, was relatively highly expressed in epithelial cell-rich organs such as mammary glands, lung, and kidney in mice.However, knockdown of miR-200a prevented increases in ß-casein and E-cadherin mRNA expression.Protein analysis revealed that E-cadherin signal was decreased and ZEB1 (a marker of EMT) was increased following miR-200a knockdown.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Physiology, Department of Veterinary Medicine, Tokyo University of Agriculture and Technology, Tokyo, Japan. nagaokak@cc.tuat.ac.jp

ABSTRACT
Mammary gland epithelial cells undergo periodic cycles of proliferation, differentiation, and involution. Many studies have reported that miRNAs, which are small, non-coding RNAs, influence a variety of biological processes during posttranscriptional regulation. Here, we found that one miRNA, miR-200a, was relatively highly expressed in epithelial cell-rich organs such as mammary glands, lung, and kidney in mice. In mammary glands, miR-200a expression increased during mid-pregnancy through lactation; its expression was stimulated by lactogenic hormone treatment of mammary epithelial cells. Lactogenic hormone also induced the expression of milk protein ß-casein mRNA (a marker of cell differentiation) and E-cadherin mRNA (a marker of epithelial cells). However, knockdown of miR-200a prevented increases in ß-casein and E-cadherin mRNA expression. Protein analysis revealed that E-cadherin signal was decreased and ZEB1 (a marker of EMT) was increased following miR-200a knockdown. Finally, in a three-dimensional culture system modeling lumen-containing mammary ducts, miR-200a knockdown decreased the cavity formation rate and suppressed claudin-3 and par-6b expression, indicating reduced epithelial cell polarity. These observations suggest that miR-200a is important for maintaining the epithelial cell phenotype, which contributes to lactogenic hormone induction of cellular differentiation in mammary glands.

Show MeSH
Related in: MedlinePlus