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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.