title: Pseudolaric acid B induced cell cycle arrest, autophagy and senescence in murine fibrosarcoma l929 cell.
abstract: OBJECTIVE
  PAB induced various cancer cell apoptosis, cell cycle arrest and senescence. But in cell line murine fibrosarcoma L929, PAB did not induce apoptosis, but autophagy, therefore it was thought by us as a good model to research the relationship of cell cycle arrest, autophagy and senescence bypass apoptosis.
  
  
  METHODS
  Inhibitory ratio was assessed by 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) analysis. Phase contrast microscopy visualized cell morphology. Hoechst 33258 staining for nuclear change, propidium iodode (PI) staining for cell cycle, monodansylcadaverine (MDC) staining for autophagy, and rodanmine 123 staining for mitochondrial membrane potential (MMP) were measured by fluorescence microscopy or flowcytometry. Apoptosis was determined by DNA ladder test. Protein kinase C (PKC) activity was detected by PKC assay kit. SA-β-galactosidase assay was used to detect senescence. Protein expression was examined by western blot.
  
  
  RESULTS
  PAB inhibited L929 cell growth in time-and dose-dependent manner. At 12 h, 80 μmol/L PAB induced obvious mitotic arrest; at 24 h, PAB began to induce autophagy; at 36 h, cell-treated with PAB slip into G1 cell cycle; and 3 d PAB induced senescence. In time sequence PAB induced firstly cell cycle arrest, then autophagy, then slippage into G1 phase, lastly senescence. Senescent cells had high level of autophagy, inhibiting autophagy led to apoptosis, and no senescence. PAB activated PKC activity to induce cell cycle arrest, autophagy and senescence, inhibiting PKC activity suppressed cell cycle arrest, autophagy and senescence.
  
  
  CONCLUSIONS
  PAB induced cell cycle arrest, autophagy and senescence in murine fibrosarcoma L929 cell through PKC.
authors: Institute of virology and AIDS research, The first hospital of Jilin University, 519 Dongminzhu Street, Changchun 130021, P R China.
authors: Jing hua
authors: Jh
authors: Yu
authors: Chun yu
authors: Cy
authors: Liu
authors: Gui bin
authors: Gb
authors: Zheng
authors: Li Ying
authors: LY
authors: Zhang
authors: Ming hui
authors: Mh
authors: Yan
authors: Wen yan
authors: Wy
authors: Zhang
authors: Xian ying
authors: Xy
authors: Meng
authors: Xiao fang
authors: Xf
authors: Yu
mesh_terms: D000818:Animals; D001343:Autophagy / Q000187:drug effects* / Q000235:genetics; D059447:Cell Cycle Checkpoints / Q000187:drug effects* / Q000235:genetics; D016922:Cellular Senescence / Q000187:drug effects*; D004224:Diterpenes / Q000008:administration & dosage; D005354:Fibrosarcoma / Q000188:drug therapy / Q000378:metabolism / Q000473:pathology*; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D051379:Mice; D011493:Protein Kinase C / Q000096:biosynthesis*
chemical_list: D004224:Diterpenes; C058391:pseudolaric acid B; D011493:Protein Kinase C
keywords: PKC; autophagy; cell cycle arrest; murine fibrosarcoma L929 cells; pseudolaric acid B (PAB); senescence.
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