title: gga-miR-26a targets NEK6 and suppresses Marek's disease lymphoma cell proliferation.
abstract: MicroRNA (miRNA) are a class of highly conserved, small noncoding RNA that emerge as key posttranscriptional regulators in various neoplastic transformations. Our previous study profiling the miRNA transcriptome in Marek's disease virus (MDV)-induced lymphoma revealed many novel and differentially expressed miRNA, including gga-miR-26a, which was downregulated in MDV-infected spleens of chickens. In this study, differential expression of gga-miR-26a between MDV-infected and noninfected spleens at 4, 7, 14, 21, and 28 d postinfection was analyzed by real-time PCR. The results showed gga-miR-26a were downregulated in MDV-infected spleens at cytolytic infection, latency, and tumor transformation phases. Subsequent cell proliferation assay revealed cell viability was lower in gga-miR-26a mimic transfection group than that in negative controls. Target genes of gga-miR-26a were identified by luciferase reporter gene assay. The results showed significant interaction between gga-miR-26a and Never In Mitosis Gene A (NIMA)-related kinase 6 (NEK6) gene. Subsequent gain of function experiment and Western blot assay showed that mRNA and protein levels of NEK6 were downregulated after gga-miR-26 mimic was transfected into MDV-transformed lymphoid cell line (MSB-1), indicating that NEK6 was modulated by gga-miR-26a. The expression of NEK6 showed a higher trend in MDV-infected samples including tumorous spleen and MD lymphoma from liver than that in noninfected controls. The results suggested that gga-miR-26a inhibited MSB-1 cell proliferation. Gga-miR-26a and its direct target, NEK6, might play important roles in MDV infection.
authors.affiliation: Department of Animal Genetics and Breeding, National Engineering Laboratory for Animal Breeding, College of Animal Science and Technology, China Agricultural University, Beijing 100193, China.
authors.forename: Xin
authors.initials: X
authors.lastname: Li
authors.forename: Ling
authors.initials: L
authors.lastname: Lian
authors.forename: Daixi
authors.initials: D
authors.lastname: Zhang
authors.forename: Lujiang
authors.initials: L
authors.lastname: Qu
authors.forename: Ning
authors.initials: N
authors.lastname: Yang
mesh_terms: D000818:Animals; D030161:Avian Proteins / Q000235:genetics* / Q000378:metabolism; D015153:Blotting, Western / Q000662:veterinary; D049109:Cell Proliferation; D002645:Chickens; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D008381:Herpesvirus 2, Gallid / Q000502:physiology; D006801:Humans; D008223:Lymphoma / Q000235:genetics / Q000503:physiopathology / Q000662:veterinary* / Q000821:virology; D008380:Marek Disease / Q000235:genetics* / Q000503:physiopathology / Q000821:virology; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D011201:Poultry Diseases / Q000235:genetics* / Q000503:physiopathology / Q000821:virology; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000662:veterinary; D013160:Splenic Neoplasms / Q000235:genetics / Q000503:physiopathology / Q000662:veterinary / Q000821:virology
chemical_list: D030161:Avian Proteins; D035683:MicroRNAs; D017346:Protein Serine-Threonine Kinases
keywords: Marek’s disease; NEK6; chicken; gga-miR-26a