title: Lentivirus-based RNA silencing of Nemo-like kinase (NLK) inhibits the CAL 27 human adenosquamos carcinoma cells proliferation and blocks G0/G1 phase to S phase.
abstract: BACKGROUND
  The Nemo-like kinase (NLK) is a serine/threonine-protein kinase that involved in a number of signaling pathways regulating cell fate. Variation of NLK has been shown to be associated with the risk of cancer. However, the function of NLK in oral adenosquamous carcinoma cells line CAL-27 is unknown.
  
  
  METHODS
  In this study, we evaluated the function of NLK in CAL-27 cells by using lentivirus-mediated RNA silence. The targeted gene expression, cell proliferation and cell cycle are investigated by RT-PCR, western-blot, MTT method, colony forming assay and flow cytometry analysis respectively.
  
  
  RESULTS
  After NLK silencing, the number of colonies was significantly reduced (54 ± 5 colonies/well compared with 262 ± 18 colonies/well in non-infected or 226 ± 4 colonies/well in negative control group (sequence not related to NLK sequence with mismatched bases). Using crystal violet staining, we also found that the cell number per colony was dramatically reduced. The RNA silencing of NLK blocks the G0/G1 phase to S phase progression during the cell cycle.
  
  
  CONCLUSIONS
  These results suggest that NLK silencing by lentivirus-mediated RNA interference would be a potential therapeutic method to control oral squamous carcinoma growth.
authors: Department of Anatomy Shandong University, School of Medicine, Jinan Shandong, 250012, P R China.
authors: Bin
authors: B
authors: Zhang
authors: Ke Yi
authors: KY
authors: Li
authors: Hai Ying
authors: HY
authors: Chen
authors: Shao Dong
authors: SD
authors: Pan
authors: Shuang Feng
authors: SF
authors: Chen
authors: Wei Feng
authors: WF
authors: Zhang
authors: Chun Peng
authors: CP
authors: Xia
authors: Li Cheng
authors: LC
authors: Jiang
authors: Xian Bin
authors: XB
authors: Liu
authors: Feng Jun
authors: FJ
authors: Zhao
authors: Dao Ying
authors: DY
authors: Yuan
authors: Le Xin
authors: le X
authors: Wang
authors: Ya Ping
authors: YP
authors: Wu
authors: Shu Wei
authors: SW
authors: Liu
mesh_terms: D018196:Carcinoma, Adenosquamous / Q000201:enzymology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D016193:G1 Phase / Q000235:genetics / Q000502:physiology*; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D016086:Lentivirus / Q000235:genetics*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism*; D034622:RNA Interference / Q000502:physiology*; D016192:Resting Phase, Cell Cycle / Q000235:genetics / Q000502:physiology*; D016196:S Phase / Q000235:genetics / Q000502:physiology*
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; C470551:NLK protein, human; D017346:Protein Serine-Threonine Kinases
keywords: Lentivirus; Nemo-like kinase (NLK); Oral Squamous Cell Carcinoma; RNAi
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