title: Cellular MicroRNA Let-7a Suppresses KSHV Replication through Targeting MAP4K4 Signaling Pathways.
abstract: BACKGROUND
  Kaposi's sarcoma (KS)-associated herpesvirus (KSHV) is the etiologic agent of KS, the most common AIDS-related malignancy. The majority of KS tumor cells harbor latent KSHV virus but only a small percentage undergoes spontaneous lytic replication. Viral reactivation from latency is crucial for the pathogenesis and development of KS, but the cellular mechanisms underlying the switch between viral latency and replication are not well understood.
  
  
  METHODS
  The level of let-7 miRNAs and MAP4K4 in KSHV infected 293T cells were quantified by real-time PCRs. Let-7 expression was silenced by the miRNA sponge technique. In let-7a transfected 293T cells, the expression of MAP4K4 was measured by real-time PCR and western blot. Luciferease expression was employed to examine the effect of let-7a on the 3'-untranslated region (UTR) of the MAP4K4 gene in 293T cells. Real-time PCR was used to quantify the KSHV copy numbers in BC-3 cells in which the expression of let-7a and/or MAP4K4 were altered. Finally, ERK, JNK and p38 protein production and their phosphorylation status were detected by western blots in let-7a or MAP4K4 transfected BCBL-1 cells.
  
  
  RESULTS
  The expression of microRNA let-7 was dramatically decreased in KSHV infected 293T cells, but that of MAP4K4 was increased significantly. Let-7a is physically associated with and targets the MAP4K4 3'UTR, and inhibits MAP4K4 expression at both mRNA and protein levels. MAP4K4 stimulates KSHV reactivation from latency, whereas let-7a inhibits the function of MAP4K4 by reversing the function of MAP4K4 on JNK, phospho-JNK and phospho-ERK1/2 levels.
  
  
  CONCLUSIONS
  Our results establish that let-7a specifically suppresses MAP4K4 expression, and further inhibits KSHV reactivation by interfering with the function of MAP4K4 on the MAPK pathway, highlighting let-7a as a potential treatment for KS.
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Xiaohua
authors.initials: X
authors.lastname: Tan
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Gao
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Yulong
authors.initials: Y
authors.lastname: Nan
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Jinxia
authors.initials: J
authors.lastname: Zhang
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Chunhong
authors.initials: C
authors.lastname: Di
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Wang
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Fuzhi
authors.initials: F
authors.lastname: Lian
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Yifei
authors.initials: Y
authors.lastname: Cao
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Hu
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Liangwen
authors.initials: L
authors.lastname: Xu
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Haiyan
authors.initials: H
authors.lastname: Ma
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Hong
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Tingjie
authors.initials: T
authors.lastname: Liu
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Yinyin
authors.initials: Y
authors.lastname: Wu
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Xianrong
authors.initials: X
authors.lastname: Xu
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Yutao
authors.initials: Y
authors.lastname: Yan
authors.affiliation: School of Medicine, Hangzhou Normal University, Hangzhou, Zhejiang, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Yang
mesh_terms: D002460:Cell Line; D005786:Gene Expression Regulation; D019288:Herpesvirus 8, Human / Q000502:physiology*; D054884:Host-Pathogen Interactions; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics* / Q000378:metabolism; D020935:MAP Kinase Signaling System; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics* / Q000378:metabolism; D012514:Sarcoma, Kaposi / Q000235:genetics* / Q000378:metabolism / Q000503:physiopathology; D014775:Virus Activation; D014779:Virus Replication
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D035683:MicroRNAs; C487931:mirnlet7 microRNA, human; C117090:MAP4K4 protein, human; D017346:Protein Serine-Threonine Kinases