Han Bo, Lian Ling, Li Xin, Zhao Chunfang, Qu Lujiang, Liu Changjun, Song Jiuzhou, Yang Ning
Department of Animal Genetics and Breeding, National Engineering Laboratory for Animal Breeding College of Animal Science and Technology, China Agricultural University, Beijing, 100193, China.
Division of Avian Infectious Diseases, Harbin Veterinary Research Institute of Chinese Academy of Agricultural Sciences, Harbin, 150001, China.
Mol Biol Rep. 2016 Jul;43(7):667-76. doi: 10.1007/s11033-016-4002-2. Epub 2016 May 13.
Marek's disease (MD) is an infectious disease of chickens caused by MD virus (MDV), which is a herpesvirus that initiates tumor formation. Studies have indicated that microRNAs (miRNAs) are linked with the development of cancers or tumors. Previously, gga-miR-130a was discovered downregulated in MDV-infected tissues. Here, we aimed to explore the further function of gga-miR-130a in MD. The expression of gga-miR-130a in MDV-infected and uninfected spleens was detected by quantitative real-time PCR (qRT-PCR). Subsequently, proliferation and migration assays of MDV-transformed lymphoid cells (MSB1) were carried out by transfecting gga-miR-130a. The target genes of gga-miR-130a were predicted using TargetScan and miRDB and clustered through Gene Ontology analysis. The target genes were validated by western blot, qRT-PCR, and a dual luciferase reporter assay. Our results show that the expression of gga-miR-130a was reduced in MDV-infected spleens. Gga-miR-130a showed an inhibitory effect on MSB1 cell proliferation and migration. Two target genes, homeobox A3 (HOXA3) and MyoD family inhibitor domain containing (MDFIC), were predicted and clustered to cell proliferation. Results indicate that gga-miR-130a regulates HOXA3 and MDFIC at the protein level but not at the mRNA level. Moreover, the gga-miR-130a binding sites of two target genes have been confirmed. We conclude that gga-miR-130a can arrest MSB1 cell proliferation and migration, and target HOXA3 and MDFIC, which are both involved in the regulation of cell proliferation. Collectively, gga-miR-130a plays a critical role in the tumorigenesis associated with chicken MD.
马立克氏病(MD)是一种由马立克氏病病毒(MDV)引起的鸡传染性疾病,MDV是一种引发肿瘤形成的疱疹病毒。研究表明,微小RNA(miRNA)与癌症或肿瘤的发展有关。此前,发现gga-miR-130a在MDV感染的组织中表达下调。在此,我们旨在探索gga-miR-130a在MD中的进一步功能。通过定量实时PCR(qRT-PCR)检测gga-miR-130a在MDV感染和未感染脾脏中的表达。随后,通过转染gga-miR-130a对MDV转化的淋巴细胞(MSB1)进行增殖和迁移分析。使用TargetScan和miRDB预测gga-miR-130a的靶基因,并通过基因本体分析进行聚类。通过蛋白质印迹、qRT-PCR和双荧光素酶报告基因检测验证靶基因。我们的结果表明,gga-miR-130a在MDV感染的脾脏中表达降低。gga-miR-130a对MSB1细胞增殖和迁移具有抑制作用。预测了两个靶基因,即同源盒A3(HOXA3)和含肌分化因子抑制结构域(MDFIC),并聚类到细胞增殖相关基因。结果表明,gga-miR-130a在蛋白质水平而非mRNA水平上调节HOXA3和MDFIC。此外,已证实两个靶基因的gga-miR-130a结合位点。我们得出结论,gga-miR-130a可阻止MSB1细胞增殖和迁移,并靶向HOXA3和MDFIC,这两个基因均参与细胞增殖的调节。总体而言,gga-miR-130a在与鸡MD相关的肿瘤发生中起关键作用。