Li Hui, Zheng Huiling, Sun Yongsen, Yu Qian, Li Lihui
College of Animal Science and Technology, Northwest A&F University, Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.
College of Animal Science and Technology, Northwest A&F University, Key Laboratory of Molecular Biology for Agriculture, Yangling, Shaanxi 712100, China.
Gene. 2014 Nov 10;551(2):279-89. doi: 10.1016/j.gene.2014.09.005. Epub 2014 Sep 4.
The signal transducer and activator of transcription 5a (Stat5a) modulates genes involved in proliferation and survival and plays pivotal roles in regulating the function of the mammary gland during pregnancy, lactation, and involution. However, there is little information about the effects of Stat5a on apoptosis of goat mammary gland epithelial cells (GMECs). In addition, parathyroid hormone-related protein (PTHrP) is a key regulator in cellular calcium transport, mammary gland development and breast tumor biology. This study aimed to explore the interaction of Stat5a and PTHrP in GMEC apoptosis. Quantitative real time PCR (qRT-PCR) suggested that Stat5a was predominantly expressed in the mammary gland, lung, liver and spleen of goats. Treating the GMECs with pimozide, an inhibitor of Stat5a that decreases Stat5a tyrosine phosphorylation, increased PTHrP levels in GMECs in a dose-dependent manner and simultaneously promoted apoptosis of the GMECs. We also demonstrated that PTHrP inhibition induced GMEC apoptosis and restrained cell proliferation. In contrast, PTHrP overexpression protected GMECs from pimozide- and calcium-induced apoptosis, and promoted cell proliferation. Furthermore, pimozide and CaCl2 downregulated the antiapoptotic protein Bcl-2 mRNA expression, respectively, and these effects were protected by PTHrP overexpression. Interestingly, we also found that Stat5a suppressed the expression of matrix metalloproteinase 9 (MMP-9) which can induce goat mammary epithelial cell migration, but PTHrP increased MMP-9 mRNA level. Thus, Stat5a may regulate GMEC survival by regulating the expression of PTHrP.
信号转导与转录激活因子5a(Stat5a)可调节参与细胞增殖和存活的基因,并在妊娠、泌乳和退化过程中调节乳腺功能方面发挥关键作用。然而,关于Stat5a对山羊乳腺上皮细胞(GMECs)凋亡影响的信息却很少。此外,甲状旁腺激素相关蛋白(PTHrP)是细胞钙转运、乳腺发育和乳腺肿瘤生物学中的关键调节因子。本研究旨在探讨Stat5a与PTHrP在GMECs凋亡中的相互作用。定量实时PCR(qRT-PCR)表明,Stat5a在山羊的乳腺、肺、肝和脾中主要表达。用匹莫齐特(一种可降低Stat5a酪氨酸磷酸化水平的Stat5a抑制剂)处理GMECs,可使GMECs中的PTHrP水平呈剂量依赖性增加,并同时促进GMECs的凋亡。我们还证明,抑制PTHrP可诱导GMECs凋亡并抑制细胞增殖。相反,过表达PTHrP可保护GMECs免受匹莫齐特和钙诱导的凋亡,并促进细胞增殖。此外,匹莫齐特和CaCl2分别下调抗凋亡蛋白Bcl-2的mRNA表达,而过表达PTHrP可保护这些作用。有趣的是,我们还发现Stat5a可抑制基质金属蛋白酶9(MMP-9)的表达,MMP-9可诱导山羊乳腺上皮细胞迁移,但PTHrP可增加MMP-9的mRNA水平。因此,Stat5a可能通过调节PTHrP的表达来调节GMECs的存活。