title: Inhibition Mechanism of Novel Pyrazolo[1,5-a]pyrazin-4(5H)-one Derivatives Against Proliferation of A549 and H322 Cancer Cells.
abstract: OBJECTIVE
  To explore the inhibition mechanism and safety of pyrazolo[1,5-a]pyrazin-4(5H)-one derivatives against proliferation of human lung cancer A549 cells, H322 cells, and human umbilical vein endothelial cell (HUVEC).
  
  
  METHODS
  Cells were treated with 40 Μmol/L of the ppo3a, ppo3b, ppo3i, and 0.1% DMSO (control) for 48 hours, respectively. Apoptosis was determined by Hoechst 33258 staining assay in H322 and A549 cells. Cell cycle distribution was determined by flow cytometry analysis in A549 cell. LC3-II, p53, and heat shock protein (HSP) 70 protein levels were detected by Western blotting in A549 cells treated with ppo3b for 48 hours. The morphology and viability of HUVEC were observed by inverted microscope and sulforhodamine B (SRB) assay.
  
  
  RESULTS
  Ppo3a, ppo3b, and ppo3i significantly induced apoptosis in H322 and A549 cells. A strong G1-phase arrest was concomitant with the growth inhibitory effect on A549 cells. Ppo3b effectively elevated the p53 protein level, but significantly reduced the HSP70 protein level. There were no significantly inhibitory effect on the morphology and viability of HUVEC when treated with ppo3a, ppo3b, and ppo3i.
  
  
  CONCLUSIONS
  ppo3a, ppo3b, and ppo3i could inhibit H322 proliferation through apoptosis and inhibit A549 through apoptosis and G1-phase arrest. The protein p53 and HSP70 might involve in the inhibition effects. These derivatives might be a clue to find effective and safe drug for lung cancers.
authors.affiliation: 1. Department of Medical Morphology, School of Medicine, Hubei University of Art and Science, Xiangyang 441053, Hubei, China; 2. Shandong Provincial Key Laboratory of Animal Cells and Developmental Biology, School of Life Science, Shandong University, Jinan 250100, China;
authors.forename: Jin-hui
authors.initials: JH
authors.lastname: Shao
authors.affiliation: Department of Medical Morphology, School of Medicine, Hubei University of Art and Science, Xiangyang 441053, Hubei, China.
authors.forename: Gui-hua
authors.initials: GH
authors.lastname: Feng
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000032:analysis / Q000502:physiology; D006801:Humans; D011720:Pyrazoles / Q000494:pharmacology*; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000502:physiology
chemical_list: D018840:HSP70 Heat-Shock Proteins; D011720:Pyrazoles; D016159:Tumor Suppressor Protein p53