Gga-miR-29b-3p抑制MDV转化细胞侵袭和原癌基因Meq表达

Gga-miR-29b-3p Inhibit Invasion and the Expression of Oncogene Meq in MDV-transformed Cells

目的:研究gga-miR-29b-3p对马立克氏病(MD)肿瘤转化细胞侵袭和原癌基因Meq表达的影响。方法:选取SPF白来航鸡在感染马立克氏病病毒(MDV)后引发的内脏淋巴瘤为样本,通过实时荧光定量PCR检测gga-miR-29b-3p的表达情况;利用在线生物软件对gga-miR-29b-3p潜在的靶基因进行功能分析。以该miRNA为研究对象,MDV转化细胞系MDCC-MSB1为试验材料,分别转染miRNA激动剂或阴性对照,检测细胞侵袭相关基因MMP2和MMP9以及MDV原癌基因Meq的表达情况。结果:Gga-miR-29b-3p在感染MDV白来航鸡的肿瘤化脾脏和肝脏淋巴瘤中均显著低表达。信号通路方面miRNA的预测靶基因分别参与FoxO信号通路、mTOR信号通路、Jak-STAT信号通路、Toll样受体信号通路、ErbB信号通路、VEGF信号通路和细胞凋亡等,这些信号通路可参与肿瘤发生进程。转染miRNA激动剂后,MMP2和MMP9基因的表达量在48 h显著下调(P<0.05);Meq基因在48 h表达显著下调(P<0.05)。结论:Gga-miR-29b-3p抑制细胞侵袭和病毒原癌基因的表达,其潜在靶基因能够介导肿瘤发生,提示该miRNA可能参与MD肿瘤转化过程。

Objective:To reveal the effects of gga-miR-29b-3p on invasion and the expression of oncogene Meq in Marek's disease virus(MDV)-transformed MSB1.Methods:Visceral lymphoma caused by MDV infection in White Leghorn chickens specific pathogen free was selected and it was used for the detection of gga-miR-29b-3p through quantitative real-time PCR(qRT-PCR).The potential target genes of gga-miR-29b-3p were analyzed by online biological software.Meanwhile,we chose this miRNA as the research object,and the MDV-transformed cell line MDCC-MSB1 was used as the research material.The miRNA agomir or negative control(NC)was transfected respectively to detect the expression of cell invasion-related genes MMP 2 and MMP 9 as well as MDV proto-oncogene Meq.Results:Gga-miR-29b-3p was significantly lower expressed in tumorous spleen and lymphoma from liver infected with MDV.Its predicted target genes were involved in the FoxO signaling pathway,mTOR signaling pathway,Jak-STAT signaling pathway,Toll-like receptor signaling pathway,ErbB signaling pathway,VEGF signaling pathway and apoptosis.These signaling pathways could participate in tumorigenesis.Moreover,after transfection with agomir,the expressions of MMP 2 and MMP 9 genes were extremely significantly down-regulated at 48 h compared with those transfected with NC(P<0.05).The expression of Meq gene was significantly down-regulated at 48 h post agomir transfection(P<0.05).Conclusion:Gga-miR-29b-3p could inhibit cell invasion and viral proto-oncogene expression,and its potential target genes could mediate tumorigenesis.These evidence suggest that this miRNA may be involved in the MD tumor transformation process.