title: [P2Y purinergic receptor activated PI-3K/Akt signaling pathway in regulation of growth and invasion of prostatic cancer].
abstract: OBJECTIVE
  To investigate P2Y purinergic receptor activated PI-3K/Akt signaling pathway in the regulation of growth and invasion of prostate cancer in vitro.
  
  
  METHODS
  Western blot was used to detect phosphorylation of Akt (a downstream target molecule of PI-3K) by P2Y receptor agonist in 1E8 cells (a highly metastatic subclone derived from PC-3 prostatic cancer cell line). Cell counts, flow cytometry, Matrigel invasion assay, wound healing assay and gelatin zymography were used to detect changes of biological behaviors of 1E8 cells after P2Y receptor activation.
  
  
  RESULTS
  AMP-PNP, one non-hydrolysis ATP analogue and P2Y receptor agonist, induced significant phosphorylation of Akt in a time- and dose-dependent manner in IE8 cells. LY294002, a specific inhibitor of PI-3K, effectively blocked Akt phosphorylation induced by AMP-PNP. Continuous exposure to AMP-PNP induced significant growth inhibition of 1E8 cells (inhibition rate at 50.2% at the 8th day), and this inhibition was mainly due to an arrest at S phase of the cell cycle (the S phase fraction of AMP-PNP treated cells was 22.3% higher than that of the control). Application of LY294002 did not reverse the growth inhibition effect of AMP-PNP. Matrigel invasion assay showed that AMP-PNP stimulation increased invasive ability of 1E8 cells, and this effect was effectively blocked by LY294002. No significant changes in the activation of MMP-2 and MMP-9 were detected by gelatin zymography, although wound healing assay showed 21.2% increase in cell migration after AMP-PNP treatment.
  
  
  CONCLUSIONS
  PI-3K/Akt signaling pathway participates in P2Y receptor-stimulated prostate cancer invasion by enhancing cell motility, rather than up-regulating MMP-2 and MMP-9 activities. PI-3K signaling pathway plays an important role in prostate cancer proliferation, but is not involved in P2Y receptor mediated growth inhibition.
authors.affiliation: Department of Pathology, Peking University Health Science Center, Beijing 100083, China.
authors.forename: Yu-xiang
authors.initials: YX
authors.lastname: Wang
authors.forename: Yong-hong
authors.initials: YH
authors.lastname: Shi
authors.forename: Li-hua
authors.initials: LH
authors.lastname: Gong
authors.forename: Yan
authors.initials: Y
authors.lastname: Li
authors.forename: Wan-jie
authors.initials: WJ
authors.lastname: Heng
authors.forename: Jiang-feng
authors.initials: JF
authors.lastname: You
authors.forename: Hao-hao
authors.initials: HH
authors.lastname: Zhong
authors.forename: Wei-gang
authors.initials: WG
authors.lastname: Fang
mesh_terms: D000266:Adenylyl Imidodiphosphate / Q000494:pharmacology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002867:Chromones / Q000494:pharmacology; D006801:Humans; D008297:Male; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D051379:Mice; D008819:Mice, Nude; D009025:Morpholines / Q000494:pharmacology; D009361:Neoplasm Invasiveness; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D000081082:Phosphoinositide-3 Kinase Inhibitors; D010766:Phosphorylation; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D058910:Purinergic P2 Receptor Agonists; D016196:S Phase / Q000187:drug effects; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D000266:Adenylyl Imidodiphosphate; D002867:Chromones; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D009025:Morpholines; D019869:Phosphatidylinositol 3-Kinases; D000081082:Phosphoinositide-3 Kinase Inhibitors; D051057:Proto-Oncogene Proteins c-akt; D058910:Purinergic P2 Receptor Agonists; C085911:2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one; C522361:MMP2 protein, human