摘要
旨在探究泌乳早期高表达lncRNA EPB41L4A-AS在奶牛乳腺上皮细胞(BMECs)中的功能及其调控机制。本研究通过将EPB41L4A-AS siRNA和si-NC分别转染BMECs,利用EdU和MTT试验检测其对BMECs增殖的影响;qPCR和Western blot试验检测ErbB3和PIK3R1、AKT、STAT5a基因表达情况。EdU和MTT结果表明,降低EPB41L4A-AS的表达后极显著提高了奶牛乳腺上皮细胞的活力(P<0.001),EdU阳性细胞数目明显增多;qPCR和Western blot结果表明,降低EPB41L4A-AS的表达后ErbB3和PIK3R1、AKT、STAT5a mRNA的相对丰度都显著高于对照组(P<0.05),且ErbB3和PIK3R1、AKT、p-AKT、STAT5a蛋白水平的相对表达量显著升高(P<0.05)。本研究结果表明,降低EPB41L4A-AS表达后能够促进ErbB3的表达,激活PI3K/AKT和JAK-STAT信号通路从而促进BMECs的增殖。本研究揭示了EPB41L4A-AS在BMECs增殖中的作用和调控途径,为深入探究lncRNA对BMECs的调控机理提供理论依据。
This study aimed to explore the function and regulatory mechanism of EPB41L4A-AS with high expression in early lactation in bovine mammary epithelial cells(BMECs).In this study,EPB41L4A-AS siRNA and si-NC were transfected into BMECs,respectively.The effect of EPB41L4A-AS siRNA on the proliferation of BMECs was detected by EdU and MTT assays;qPCR and Western blot methods were used to detect the expression of ErbB3,PIK3R1,AKT and STAT5a genes.EdU and MTT results showed that the activity of BMECs was significantly increased with the interference of EPB41L4A-AS expression(P<0.001)and the number of EdU positive cells obviously increased.qPCR and Western blot results showed that the relative mRNA abundance of ErbB3,PIK3R1,AKT and STAT5a were significantly higher than those of the control group after interfering EPB41L4A-AS expression(P<0.05),and the relative protein expression levels of ErbB3,PIK3R1,AKT,p-AKT and STAT5a significantly increased(P<0.05).The results of this study suggested that reducing the expression of EPB41L4A-AS promoted the expression of ErbB3,and activated the PI3K/AKT and JAK-STAT signaling pathways to promote the proliferation of BMECs.This study revealed the role and regulatory pathway of EPB41L4A-As in BMECs proliferation,which provided a theoretical basis for further exploring the regulation mechanism of lncRNA on BMECs.
作者
刘丽华
钟震宇
郑玉杰
王昕
LIU Lihua;ZHONG Zhenyu;ZHENG Yujie;WANG Xin(College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2022年第7期2152-2159,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金面上项目(31972556)。
