title: [Inhibitory effect of lentiviral-mediated RNA on the expression of Foxp3 protein in melanoma cells].
abstract: OBJECTIVE
  To explore the feasibility of RNA interference in the treatment of melanoma by inhibiting the Foxp3 gene expression in mouse B16 melanoma cells using RNA interference (RNAi) in vitro.
  
  
  METHODS
  Small interfering  RNA (siRNA) was designed according to Foxp3 gene. A short hairpin RNA (shRNA) lentivirus expression vector was constructed and   transfected into mouse B16 cells,  and RNA interference was induced in vitro. Western blot and real-time RT-PCR were performed  to detect the expression of Foxp3 gene. ELISA was applied to detect the changes of TGF-β(1);,  TGF-β(2);,  IL-10 and other cytokines. The B16 cells after interference were co-cultured with CD4(+);CD25(-);T lymphocytes. CCK8 assay was used to monitor the proliferation of CD4(+);CD25(-);T lymphocytes.
  
  
  RESULTS
  shRNA could suppress the expression level of Foxp3, down-regulate the inhibitory ability of tumor cells on the proliferation of CD4(+);CD25(-);T lymphocytes,  and reduce the secretion of TGF-β(1);, TGF-β(2);,  IL-10 and other cytokines,  in particular the expression of TGF-β(2);.
  
  
  CONCLUSIONS
  RNA interference can inhibit the expression of target gene Foxp3 in mice melanoma cells and the proliferation of tumor cells. It can also reduce the inhibition on the proliferation of CD4(+);CD25(-);T lymphocytes,  and the secretion of inhibitory cytokines.
authors.affiliation: The Second Department of Oncology, The First Affiliated Hospital of Chinese PLA General Hospital, Beijing 100048, China. dianjunchen@yahoo.cn
authors.forename: Dian-Jun
authors.initials: DJ
authors.lastname: Chen
authors.forename: Xiao-Song
authors.initials: XS
authors.lastname: Li
authors.forename: Hui
authors.initials: H
authors.lastname: Zhao
authors.forename: Xiu-Li
authors.initials: XL
authors.lastname: Shi
authors.forename: Huan-Huan
authors.initials: HH
authors.lastname: Zhang
authors.forename: Zhong-Yi
authors.initials: ZY
authors.lastname: Fan
authors.forename: Yong-Ming
authors.initials: YM
authors.lastname: Yao
authors.forename: Nan
authors.initials: N
authors.lastname: DU
mesh_terms: D000818:Animals; D015153:Blotting, Western; D015496:CD4-Positive T-Lymphocytes / Q000166:cytology / Q000378:metabolism; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002478:Cells, Cultured; D002766:Cholecystokinin / Q000378:metabolism; D018920:Coculture Techniques; D051858:Forkhead Transcription Factors / Q000235:genetics* / Q000378:metabolism; D015972:Gene Expression Regulation, Neoplastic; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D008545:Melanoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D051379:Mice; D008856:Microscopy, Fluorescence; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D053773:Transforming Growth Factor beta1 / Q000378:metabolism; D053781:Transforming Growth Factor beta2 / Q000378:metabolism
chemical_list: D051858:Forkhead Transcription Factors; C498833:Foxp3 protein, mouse; D034741:RNA, Small Interfering; D053773:Transforming Growth Factor beta1; D053781:Transforming Growth Factor beta2; D016753:Interleukin-10; D049452:Green Fluorescent Proteins; D002766:Cholecystokinin