title: [Inhibitory effect of NaV1.9 gene silencing on proliferation, phagocytosis and migration in RAW264.7 cells].
abstract: OBJECTIVE
  To establish the cell line with stable voltage-gated sodium channels (VGSCs/NaVs) α subunit NaV1.9 gene silencing through RNA interference (RNAi) in murine RAW264.7 macrophages, and to investigate proliferation, phagocytosis and migration in this cell line.
  
  
  METHODS
  The stable NaV1.9-deficient cell line was generated by selection in G418 after the transfection of short hairpin (shRNA) plasmid with Lipofectamine TM2000. RNAi efficiency was qualified by RT-PCR; proliferation ability was measured by CCK-8 assay; cell cycle and phagocytic ability were analyzed by flow cytometry; and migrating ability was detected by Transwell migration assay.
  
  
  RESULTS
  Stable NaV1.9-deficient cell line was established and the expression of NaV1.9 was reduced by 80%. CCK-8 assay and flow cytometry showed that the proliferation of the NaV1.9-deficient cell line was inhibited (P<0.05). Flow cytometry revealed that phagocytic ability was reduced in the cell line (P<0.05). Transwell migration assay demonstrated that migrating ability was depressed in the cell line (P<0.05).
  
  
  CONCLUSIONS
  In the stable NaV1.9-deficient cells we successfully constructed, proliferation, phagocytosis and migration were obviously inhibited.
authors: Tianjin Medical University, Tianjin, China. ycluo.cardiolab@gmail.com
authors: Yuechen
authors: Y
authors: Luo
authors: Xin
authors: X
authors: Zhou
authors: Wenjie
authors: W
authors: Ji
authors: Haiying
authors: H
authors: Sun
authors: Ruiyi
authors: R
authors: Lu
authors: Tiemin
authors: T
authors: Jiang
authors: Yuming
authors: Y
authors: Li
mesh_terms: D000818:Animals; D002453:Cell Cycle / Q000235:genetics; D002460:Cell Line; D002465:Cell Movement / Q000235:genetics*; D049109:Cell Proliferation; D020868:Gene Silencing; D008264:Macrophages / Q000166:cytology*; D051379:Mice; D062558:NAV1.9 Voltage-Gated Sodium Channel / Q000172:deficiency* / Q000235:genetics* / Q000378:metabolism; D010587:Phagocytosis / Q000235:genetics*
chemical_list: D062558:NAV1.9 Voltage-Gated Sodium Channel; C491885:Scn11a protein, mouse