title: [Cannabinoid WIN55, 212-2 inhibits proliferation, invasion and migration of human SMMC-7721 hepatocellular carcinoma cells].
abstract: OBJECTIVE
  To investigate the effects of WIN55, 212-2 (WIN) on the proliferation, invasion and migration of SMMC-7721 hepatocellular carcinoma cells and its underlying mechanisms.
  
  
  METHODS
  SMMC-7721 cells were treated with (0, 1, 5, 10, 20) μmol/L WIN, and cell viability was determined by CCK-8 assay. The morphological changes of the cells were observed under a fluorescence microscope with Hoechst33258 staining. Cell apoptosis was measured by flow cytometry combined with annexin V-FITC/PI staining. The expression levels of apoptosis-related proteins P53, P21, Bcl-2 and Bax, and the phosphorylated AKT (p-AKT) and phosphorylated extracellular signal-regulated kinase (p-ERK) were analyzed by Western blotting. Transwell(TM) invasion assay was used to detect cell invasion ability. Would healing assay was performed to test cell migration ability. The expression level of matrix metalloproteinase 14 (MMP-14) was evaluated by Western blotting.
  
  
  RESULTS
  WIN inhibited the proliferation of SMMC-7721 cells and induced cell apoptosis in a dose-dependent manner. After treatment with WIN, the cell nucleus concentrated and broken, indicating obvious cell apoptosis. Western blotting exhibited an up-regulation in the protein expression of P53, P21 and Bax. And the anti-apoptotic protein Bcl-2 was repressed. The expression levels of AKT, p-AKT and p-ERK were down-regulated, whereas the expression of total ERK was not obviously changed. Compared with control group, there was a significant inhibition of cell invasion and migration abilities when SMMC-7721 cells were treated with WIN. The expression level of MMP-14 decreased as well.
  
  
  CONCLUSIONS
  WIN can inhibit the proliferation of SMMC-7721 cells and induce cell apoptosis. The mechanism is associated with the activation of P53 and the inhibition of AKT, p-AKT and p-ERK. WIN can inhibit the invasion and migration of SMMC-7721 cells through down-regulating the protein expression of MMP-14.
authors.affiliation: Department of Biochemistry, Guangzhou Medical University, Guangzhou 511436, China.
authors.forename: Jianglin
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Biochemistry, Guangzhou Medical University, Guangzhou 511436, China.
authors.forename: Yuanfei
authors.initials: Y
authors.lastname: Deng
authors.affiliation: Department of Biochemistry, Guangzhou Medical University, Guangzhou 511436, China.
authors.forename: Yin
authors.initials: Y
authors.lastname: Tan
authors.affiliation: 2011 Undergraduate Student, Department of Biotechnology, Guangzhou Medical University, Guangzhou 511436, China.
authors.forename: Kefa
authors.initials: K
authors.lastname: Li
authors.affiliation: 2011 Undergraduate Student, Department of Biotechnology, Guangzhou Medical University, Guangzhou 511436, China.
authors.forename: Boguan
authors.initials: B
authors.lastname: Wen
authors.affiliation: Department of Biochemistry, Guangzhou Medical University, Guangzhou 511436, China. *Corresponding author, E-mail: zhaoqing66@yahoo.com.
authors.forename: Qing
authors.initials: Q
authors.lastname: Zhao
mesh_terms: D048588:Benzoxazines / Q000494:pharmacology*; D006528:Carcinoma, Hepatocellular / Q000737:chemistry / Q000188:drug therapy* / Q000473:pathology; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D006801:Humans; D008113:Liver Neoplasms / Q000737:chemistry / Q000188:drug therapy* / Q000473:pathology; D009025:Morpholines / Q000494:pharmacology*; D009281:Naphthalenes / Q000494:pharmacology*; D009361:Neoplasm Invasiveness; D051057:Proto-Oncogene Proteins c-akt / Q000037:antagonists & inhibitors / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000032:analysis
chemical_list: D048588:Benzoxazines; D009025:Morpholines; D009281:Naphthalenes; D016159:Tumor Suppressor Protein p53; C070417:(3R)-((2,3-dihydro-5-methyl-3-((4-morpholinyl)methyl)pyrrolo-(1,2,3-de)-1,4-benzoxazin-6-yl)(1-naphthalenyl))methanone; D051057:Proto-Oncogene Proteins c-akt; D048049:Extracellular Signal-Regulated MAP Kinases