title: [Role of PI3K-AKT signaling pathway in CD40-mediated proliferation and invasiveness of lung cancer cell].
abstract: OBJECTIVE
  To analyze the CD40-mediated in vitro proliferation and invasiveness of lung cancer cell H1299, and explore the role of phosphatidylinositol-3-kinase-protein kinase B (PI3K-AKT) signaling pathway in the above-mentioned process.
  
  
  METHODS
  Cell counting kit-8 (CCK-8) assay was used to detect the proliferation of tumor cells. And scratch test and Transwell chambers were applied to detect the migration and invasiveness of H1299. Western blot was utilized to examine the expressions of AKT and p-AKT and enzyme-linked immunosorbent assay (ELISA) for the CD40-mediated secretion of MMP-9.
  
  
  RESULTS
  According to the CCK-8 test, the absorbance of 5C11 groups at different timepoints was significantly higher than the control group (1.72 ± 0.18), especially when the concentration was 10 mg/L at 96 hour (2.75 ± 0.35, P < 0.05). In scratch test, the scratching width of 5C11 groups was significantly shorter than control group (1.62 ± 0.32 mm), especially when the concentration was 10 mg/L (0.05 ± 0.01 mm, P < 0.05). Western blot showed that the level of p-AKT protein increased at 10 minutes after activating by the signal of CD40 on H1299 cells. It was higher (0.65 ± 0.28) than the un-activating group (0.33 ± 0.16) at 30 min (P < 0.05). Upon the addition of LY294002, 5C11-mediated up-regulation of p-AKT was inhibited. In Transwell invasion assay, the number of cells on membranes in 5C11 group (168 ± 31) were significantly higher than the control group (105 ± 19, P < 0.05) and there was no significant difference between the 5C11-added LY294002 group (118 ± 21) and the control group (P < 0.05). The expression of MMP-9 of 5C11 group (192 ± 31 ng/L) was higher than that of the control group (110 ± 23 ng/L, P < 0.05) and no significant difference existed between the 5C11-added LY294002 group (120 ± 21 ng/L) and the control group (P < 0.05).
  
  
  CONCLUSIONS
  The in vitro proliferation and invasiveness of lung cancer H1299 may be regulated by the CD40-mediated activation of PI3K-AKT signaling pathway.
authors.affiliation: Department of Respiratory Medicine, Soochow University, Suzhou, China.
authors.forename: Yan-tian
authors.initials: YT
authors.lastname: Lü
authors.forename: Xiao-jing
authors.initials: XJ
authors.lastname: Sun
authors.forename: Cheng
authors.initials: C
authors.lastname: Chen
authors.forename: Tao
authors.initials: T
authors.lastname: Chen
authors.forename: Jian-an
authors.initials: JA
authors.lastname: Huang
mesh_terms: D023201:CD40 Ligand / Q000378:metabolism*; D045744:Cell Line, Tumor; D002465:Cell Movement; D049109:Cell Proliferation; D006801:Humans; D008175:Lung Neoplasms / Q000378:metabolism* / Q000473:pathology*; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D023201:CD40 Ligand; D058539:Phosphatidylinositol 3-Kinase; D051057:Proto-Oncogene Proteins c-akt; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9