FLCN通过AMPK-mTOR信号轴调节奶牛乳腺上皮细胞能量代谢

FLCN regulation of energy metabolism of bovine mammary epithelial cells through AMPK-MTOR signaling axis

Folliculin(FLCN)与AMPK存在交互作用并且在细胞能量代谢中具有重要作用,但乳腺上皮细胞中FLCN能量代谢调节作用尚未见相关报道。采用激光共聚焦方法检测奶牛乳腺发育各时期组织FLCN表达定位,为确定FLCN在奶牛乳腺上皮细胞中功能,将FLCN基因干扰片段及pcDNA3.1-FLCN过表达载体利用脂质体转染技术转染至乳腺上皮细胞中,同时在培养基中添加AMPK激活剂和抑制剂,采用Western blot检测细胞中FLCN干扰和过表达效果以及AMPK-mTOR信号通路相关蛋白表达情况。结果表明,FLCN在哺乳期乳腺组织中表达水平显著高于退化期乳腺组织。FLCN通过AMPK-mTOR信号轴参与能量代谢调控,FLCN与mTOR之间存在正反馈调节,FLCN与AMPK之间存在负反馈调节。在奶牛乳腺上皮细胞中,FLCN可能参与泌乳调节。

Folliculin(FLCN)interacts with AMPK and plays an important role in cell energy metabolism,but the regulatory role of FLCN in energy metabolism in breast epithelial cells has not been reported.In this study,the confocal laser confocal method was used to detect the expression and localization of FLCN in milk cow mammary epithelial cells at various stages of mammary development.In order to determine the function of FLCN in milk cow mammary epithelial cells,the interference fragment of FLCN gene and the overexpression vector pcDNA3.1-FLCN were transfected into mammary epithelial cells by liposome transfection technology.Meanwhile,AMPK activator and inhibitor were added to the culture medium,and Western blot was used to detect the interference and overexpression effects of FLCN and the expression of AMPK-mTOR signaling pathway related proteins.The results showed that the expression levels of FLCN in lactation breast tissues were significantly higher than those in degenerate breast tissues.FLCN participated in the regulation of energy metabolism through AMPK-mTOR signal axis,and there was positive feedback regulation between FLCN and mTOR,while there was negative feedback regulation between FLCN and AMPK.FLCN might be involved in lactation regulation in bovine mammary epithelial cells.