title: [Effect of angular pyranocoumarin isolated from peucedanum praeruptorum on the proliferation and apoptosis of U266 cells].
abstract: OBJECTIVE
  To investigate the effects of angular pyranocoumarin (±) -4'-O- acetyl-3'-Oangeloyl- cis- khellactone (APC) extracted from peucedanum praeruptoruon on the proliferation and apoptosis of U266 cells, and to explore its related mechanism.
  
  
  METHODS
  APC was extracted by petroleum ether technique, and its purity was tested by high performance liquid chromatography, and its chemical structure was identified by magnetic resonance spectroscopy. U266 cells were treated with APC in various concentrations (0, 10, 20, 30, 40 μg/ml）for different durations（24 and 48 h). The inhibitive effect of APC on cell growth was detected by CCK-8 method. After U266 cells were incubated with APC（0, 10, 20, 30, 40 μg/ml）for 24 h, the apoptosis of cells were observed by flow cytometry stained with Annexin Ⅴ/PI and Hochest33342; the expression levels of caspase-3, 8, ERK, p-ERK, AKT and p-AKT protein were assayed by Western blot; the expression of hTERT mRNA was measured by RT-PCR.
  
  
  RESULTS
  The purity of APC identified by magnetic resonance imaging was 98.8%. The proliferation of U266 cells was inhibited, and the apoptosis was induced in a time- and/or dose- dependent manner after treatment with APC. APC could upregulate the caspase- 8, 3 protein expression and downregulate the p- ERK, p-AKT protein expression along with the increase of APC dose. APC also could downregulate the hTERT mRNA expression.
  
  
  CONCLUSIONS
  Angular pyranocoumarin APC could inhibit the proliferation and induce the apoptosis of U266 cells. The probable mechanism might be achieved by upregulating caspase-8, 3 protein expression and downregulating p-ERK, P-AKT protein and the hTERT mRNA expression.
authors.affiliation: Department of Hematology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China.
authors.forename: Qinghong
authors.initials: Q
authors.lastname: Yu
authors.affiliation: Department of Hematology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China.
authors.forename: Li
authors.initials: L
authors.lastname: Ma
authors.affiliation: Department of Hematology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China.
authors.forename: Yiping
authors.initials: Y
authors.lastname: Shen
authors.forename: Wo
authors.initials: W
authors.lastname: Zhai
authors.affiliation: Department of Hematology, The First Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310006, China.
authors.forename: Yuhong
authors.initials: Y
authors.lastname: Zhou
mesh_terms: D019661:Apiaceae / Q000737:chemistry*; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D053181:Caspase 8 / Q000378:metabolism; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015536:Down-Regulation; D005434:Flow Cytometry; D015973:Gene Expression Regulation, Leukemic; D006801:Humans; D020935:MAP Kinase Signaling System; D009101:Multiple Myeloma; D064209:Phytochemicals / Q000494:pharmacology; D039681:Pyranocoumarins / Q000494:pharmacology*; D019098:Telomerase / Q000378:metabolism
chemical_list: D064209:Phytochemicals; D039681:Pyranocoumarins; C509186:TERT protein, human; D019098:Telomerase; C505407:CASP3 protein, human; C505530:CASP8 protein, human; D053148:Caspase 3; D053181:Caspase 8