摘要
为研究miRNA-125b在致细胞病变牛病毒性腹泻病毒(cpBVDV)感染牛肾细胞(MDBK细胞)过程中诱导细胞凋亡的机制,本研究将cpBVDV感染MDBK细胞,并将构建的pcDNA-miR-125b转染MDBK细胞作为阳性对照。采用荧光定量RT-PCR(qRT-PCR)检测细胞中miRNA-125b和B细胞淋巴瘤/白血病-2(Bcl-2)mRNA转录水平,通过流式细胞仪检测细胞凋亡率。检测结果显示,cpBVDV感染MDBK细胞12 h和24 h,细胞中miRNA-125b的转录水平分别为正常细胞对照组的2.01倍和3.85倍,Bcl-2 mRNA的转录水平分别为对照组的0.71倍和0.31倍;pcDNA-miR-125b转染MDBK细胞48 h后,Bcl-2 mRNA的转录水平为正常细胞对照组的0.42倍。细胞凋亡检测结果显示,cpBVDV感染MDBK细胞12 h和24 h,细胞的凋亡率分别为15.06%和36.43%;pcDNA3.1-miR-125b转染48 h,细胞的凋亡率为25.56%。结果表明miRNA-125b在cpBVDV感染MDBK细胞过程中能够通过抑制抗凋亡基因Bcl-2 mRNA转录水平从而诱导细胞产生凋亡。
To study the apoptosis mechanism of Madin-Darby bovine kidney (MDBK) cells induced by miRNA-125b in the process of the cells infected with cytopathic bovine viral diarrhea virus (cpBVDV), MDBK cells were infected with cpBVDV or transfected with pcDNA-miR-125b as a positive control. The transcription levels of miRNA-125b and B cell lymphomaJlewkmia-2 (Bci-2) mRNA were detected by real-time RT-PCR (qRT-PCR), and the cell apoptosis was examined by Flow Cytometry. The results showed the miRNA-125b levels in MDBK cells infected with cpBVDV at 12 hours and 24 hours were 2.01 and 3.85 times higher and the Bcl-2 mRNA levels were also 0.71 and 0.31 times higher than that in normal cells, respectively. In addition, the Bcl-2 mRNA level in MDBK cells transfected with pcDNA-miR-125b at 48 hours was 0.42 times higher compared with that innormal cells. Furthermore,The apoptosis rates of cells infected with cpBVDV at 12 hours and 24 hours were 15.06% and 36.43%, respectively; and apoptosis rate of cells transfected with pcDNA-miR-125b at 48 hours was 25.56%. In conclusion, these data demonstrated that cpBVDV infection triggered the high level transcription of miRNA-125b which induced the cells to undergo apoptosis by inhibiting transcription level of Bcl-2 mRNA.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2013年第7期530-534,共5页
Chinese Journal of Preventive Veterinary Medicine
基金
牛病毒性腹泻病毒囊膜蛋白E2与牛胚胎滋养层细胞相互作用的分子机制研究(31101816)
绵羊克隆囊胚差异表达microRNA的筛选及其功能研究(31201800)