Pten Regulates Development and Lactation in the Mammary Glands of Dairy Cows

• Published in: PloS one Vol. 9; no. 7; p. e102118
• Main Authors: Wang, Zhuoran, Hou, Xiaoming, Qu, Bo, Wang, Jie, Gao, Xuejun, Li, Qingzhang
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 10.07.2014; Public Library of Science (PLoS)
• Subjects: Aging; AKT protein; Animal lactation; Animals; Apoptosis; Biology and life sciences; Breasts; Casein; Caseins - metabolism; Cattle; Cell culture; Cell Cycle; Cell proliferation; Cell Survival - drug effects; Cyclin D1; Cytometry; Dairy cattle; Dairying; Education; Epithelial cells; Epithelial Cells - drug effects; Epithelial Cells - metabolism; Female; Flow cytometry; Gene expression; Gene Expression Regulation, Developmental - drug effects; Genes; Glands; Glucose; Glucose - pharmacology; Immunohistochemistry; Inhibition; Kinases; Laboratories; Lactation; Lactation - genetics; Lactose; Lactose - metabolism; Mammals; Mammary gland; Mammary glands; Mammary Glands, Animal - drug effects; Mammary Glands, Animal - growth & development; Mammary Glands, Animal - metabolism; MAP kinase; Metabolism; Milk; mRNA; Oils & fats; Pathways; Phosphatase; Pituitary hormones; Polymerase chain reaction; Prolactin; Prolactin - pharmacology; Protein folding; Proteins; PTEN Phosphohydrolase - genetics; PTEN Phosphohydrolase - metabolism; PTEN protein; RNA; RNA, Messenger - genetics; RNA, Messenger - metabolism; RNA, Small Interfering - metabolism; Rodents; Science; Signal transduction; Signaling; Transfection; Trends; Triglycerides; Triglycerides - metabolism; Tumor suppressor genes; Viability; Western blotting; China
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0102118

Pten is a tumor suppressor gene regulating many cellular processes, including growth, adhesion, and apoptosis. In the aim of investigating the role of Pten during mammary gland development and lactation of dairy cows, we analyzed Pten expression levels in the mammary glands of dairy cows by using western blotting, immunohistochemistry, and quantitative polymerase chain reaction (qPCR) assays. Dairy cow mammary epithelial cells (DCMECs) were used to study the function of Pten in vitro. We determined concentrations of β-casein, triglyceride, and lactose in the culture medium following Pten overexpression and siRNA inhibition. To determine whether Pten affected DCMEC viability and proliferation, cells were analyzed by CASY-TT and flow cytometry. Genes involved in lactation-related signaling pathways were detected. Pten expression was also assessed by adding prolactin and glucose to cell cultures. When Pten was overexpressed, proliferation of DCMECs and concentrations for β-casein, triglyceride, and lactose were significantly decreased. Overexpression of Pten down-regulated expression of MAPK, CYCLIN D1, AKT, MTOR, S6K1, STAT5, SREBP1, PPARγ, PRLR, and GLUT1, but up-regulated 4EBP1 in DCMECs. The Pten siRNA inhibition experiments revealed results that opposed those from the gene overexpression experiments. Introduction of prolactin (PRL) increased secretion of β-casein, triglyceride, and lactose, but decreased Pten expression levels. Introduction of glucose also increased β-casein and triglyceride concentrations, but did not significantly alter Pten expression levels. The Pten mRNA and protein expression levels were decreased 0.3- and 0.4-fold in mammary glands of lactating cows producing high quality milk (milk protein >3.0%, milk fat >3.5%), compared with those cows producing low quality milk (milk protein <3.0%, milk fat <3.5%). In conclusion, Pten functions as an inhibitor during mammary gland development and lactation in dairy cows. It can down-regulate DCMECs secretion of β-casein, triglyceride, and lactose, and plays a critical role in lactation related signaling pathways.