摘要
旨在研究马立克病病毒(Marek′s disease virus,MDV)编码的mdv1-miR-M4-5p对MDCC-MSB1细胞增殖和凋亡的影响,将mdv1-miR-M4-5p模拟物、抑制物及其阴性对照转染MDCC-MSB1细胞后48 h,采用实时荧光定量PCR(qRT-PCR)检测细胞中mdv1-miR-M4-5p、TGF-β1、Smad2以及caspase-9、caspase-3、cyt-c、cyclinD1、Bcl-2等增殖和凋亡相关分子的转录;CCK-8检测细胞增殖,流式细胞术检测细胞周期和凋亡。结果显示:与对照组相比,mdv1-miR-M4-5p模拟物转染显著上调MDCC-MSB1细胞中mdv1-miR-M4-5p、cyclinD1、Bcl-2的转录水平,下调TGF-β1、Smad2、cyt-c、caspase-9和caspase-3的表达转录,促进细胞的增殖,减少G1期细胞,增加S和G2细胞,降低细胞凋亡率。与对照组相比,mdv1-miR-M4-5p抑制物转染后MDCC-MSB1细胞的增殖、凋亡及其相关分子转录的结果与mdv1-miR-M4-5p模拟物转染后的结果相反。综上,Mdv1-miR-M4-5p可促进MDCC-MSB1细胞增殖,抑制其凋亡,这可能是通过抑制TGF-β1/Smad2信号通路来实现的。
The purpose of this experiment was to study the effects of mdv1-miR-M4-5p analog encoded by Marek′s disease virus(MDV)on proliferation and apoptosis of MDCC-MSB1 cells.In this study,mdv1-miR-M4-5p mimics,inhibitors and their negative controls were transfected into MSB1 cells for 48 h.Real-time quantitative fluorescent PCR(qRT-PCR)was used to detect the transcription of mdv1-miR-M4-5p,TGF-β1,Smad2 signaling molecules,caspase-9,caspase-3,cyt-c,cyclinD1,Bcl-2 et al other molecules related to proliferation and apoptosis in MDCC-MSB1 cells.Cell proliferation was detected by CCK-8 and cell cycle and apoptosis were detected by flow cytometry.The results showed that compared with the control group,the transcription levels of mdv1-miR-M4-5p,cyclinD 1 and Bcl-2 in MDCC-MSB1 cells transfected with mdv1-miR-M4-5p mimics was significantly up-regulated.The transcription of TGF-β1,Smad2,cyt-c,caspase-9 and caspase-3 was down-regulated,the proliferation of MDCC-MSB1 cells was promoted,the the number of G1 phase cells were decreased,the S and G2 cells were increased,and the apoptosis rate was decreased.Compared with the control group,the results of the proliferation and apoptosis of MDCC-MSB1 cells and the expression of related molecules transfected with mdv1-miR-M4-5p inhibitor were opposite to those transfected with mdv1-miR-M4-5p mimics.The results showed that Mdv1-miR-M4-5p could promote the proliferation and inhibit the apoptosis of MDCC-MSB1 cells.This may be conducted by inhibiting the TGF-β1/Smad2 signaling pathway.
作者
余祖华
高梦茹
何雷
魏颖
陈建
陈松彪
丁轲
YU Zuhua;GAO Mengru;HE Lei;WEI Ying;CHEN Jian;CHEN Songbiao;DING Ke(Laboratory of Functional Microbiology and Animal Health/The Key Laboratory of Animal Disease and Public Health,College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471023,China;Luoyang Key Laboratory of Live Carrier Biomaterial and Animal Disease Prevention and Control,Luoyang 471023,China;College of Animal Science and Technology,Henan University Institute of Science and Technology,Luoyang 471023 Xinxiang 453003,China)
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2024年第8期3678-3687,共10页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家自然科学基金(U1504308,31702207)
