FLCN对奶牛乳腺上皮细胞增殖及泌乳功能的影响

Effect of FLCN on proliferation and lactation function of bovine mammary epithelial cells

FLCN基因参与多种代谢途径和细胞过程,国内外关于FLCN在奶牛乳腺发育过程中表达及调节的研究鲜有报道。应用RNA干扰技术和质粒转染技术改变FLCN基因在奶牛乳腺上皮细胞中表达量,流式细胞仪检测细胞增殖,采用qRT-PCR和Western blot检测FLCN对泌乳相关功能基因AMPK、mTOR、CyclinD1、Caspase3和β-酪蛋白表达的影响。结果表明,FLCN正向调节mTOR磷酸化水平,促进乳蛋白合成和细胞增殖,抑制细胞凋亡,负调控能量代谢调节子AMPK。FLCN在奶牛乳腺上皮细胞中可通过mTOR信号通路调控细胞增殖及乳蛋白合成,研究对揭示FLCN调控奶牛乳腺上皮细胞增殖和泌乳具有重要作用。

FLCN participated in various metabolic pathways and cellular processes, and played an important role. However, the knowledge about expression and regulation mechanism of FLCN in the development of bovine cow mammary was limited. To reveal the relationship among FLCN and bovine mammary epithelial cells development and lactation, RNA interference technique and the plasmid transfection technique were used to change the expression of FLCN in bovine mammary epithelial cells by RNA interference or expression. The proliferation of bovine mammary epithelial cells was analyzed by Flow cytometry method. The effect of FLCN on milk protein related functional genes AMPK, mTOR,Cyclin D1, Caspase 3 and β-casein in bovine mammary epithelial cells was analyzed by qRT-PCR and Western blot, respectively. The results showed that FLCN positively regulated the phosphorylation level of mTOR, thereby promoting milk protein synthesis and cell proliferation, inhibiting apoptosis, and negatively regulating the energy metabolism regulator AMPK. The results suggested that FLCN might regulate cell proliferation and milk protein synthesis through mTOR signaling pathway in bovine mammary epithelial cells.