title: Growth inhibition of hepatocellular carcinoma Huh7 cells by Lactobacillus casei extract.
abstract: OBJECTIVE
  Lactobacillus casei (L. casei) is known to exert anti-proliferation effects on many types of cancer cells. However, the effect of L. casei on liver cancer has not been reported. Accordingly, the aim of this study was to determine the anti-cancer effect of L. casei extract on Huh7 cells.
  
  
  METHODS
  L. casei ATCC393 extract was prepared and purified. After the treatment of L. casei extract on Huh7 cells, cell viability, cell cycle arrest and cell death were analyzed by flow cytometry. The expression levels of tumor necrosis factor-α receptor 1 (TNFR1) and death receptor 3 (DR3) mRNA related with extrinsic apoptosis were assessed by reverse transcription polymerase chain reaction. Additionally, P21 and P27 cell cycle proteins as well as Caspase-3, -8, -9, phospho-Bad and Bcl-2 apoptosis proteins were analyzed by western blot analysis. To determine the effect of L. casei extract on cancer stem-like cells, we analyzed changes in side population fraction through flow cytometry.
  
  
  RESULTS
  The cell viability of Huh7 cells treated with L. casei extract was decreased by 77%, potentially owing to increases in the rates of Huh7 cells arrested in the G2/M phase (3% increase) and that underwent apoptosis (6% increase). The expression levels of TNFR1 and DR3 mRNA, as well as P21 and P27 cell cycle proteins, were increased. Meanwhile, the expressions of caspase-8, -9, phospho-Bad and Bcl-2 proteins decreased. However, in the case of side population cells, no remarkable changes were observed.
  
  
  CONCLUSIONS
  L. casei extract exerts a potent anti-tumor effect on the viability of liver cancer cells, although not on cancer stem-like cells.
authors.affiliation: Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea.
authors.forename: Dae Jong
authors.initials: DJ
authors.lastname: Han
authors.forename: Jong Bin
authors.initials: JB
authors.lastname: Kim
authors.forename: Seo Young
authors.initials: SY
authors.lastname: Park
authors.forename: Man Gil
authors.initials: MG
authors.lastname: Yang
authors.forename: Hyuncheol
authors.initials: H
authors.lastname: Kim
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D053181:Caspase 8 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D059447:Cell Cycle Checkpoints / Q000187:drug effects; D002457:Cell Extracts / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000378:metabolism; D054697:Cytostatic Agents / Q000494:pharmacology*; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D006801:Humans; D007780:Lacticaseibacillus casei / Q000737:chemistry*; D008113:Liver Neoplasms / Q000473:pathology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D053219:Receptors, Tumor Necrosis Factor, Member 25 / Q000378:metabolism; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000378:metabolism; D051025:bcl-Associated Death Protein / Q000378:metabolism
chemical_list: D002457:Cell Extracts; D050759:Cyclin-Dependent Kinase Inhibitor p21; D054697:Cytostatic Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D053219:Receptors, Tumor Necrosis Factor, Member 25; D047888:Receptors, Tumor Necrosis Factor, Type I; C506033:TNFRSF25 protein, human; D051025:bcl-Associated Death Protein; D050760:Cyclin-Dependent Kinase Inhibitor p27; D053181:Caspase 8; D053453:Caspase 9
keywords: Lactobacillus casei; cancer stem-like cell; liver cancer; proliferation; side population
references.citation: Cancer Lett. 1994 Sep 30;85(1):93-103
references.pmid: 7923109
references.citation: Exp Biol Med (Maywood). 2011 Nov;236(11):1351-9
references.pmid: 21908486
references.citation: Gan. 1981 Aug;72(4):517-23
references.pmid: 6796451
references.citation: Phys Sportsmed. 1996 Dec;24(12):35-40
references.pmid: 20086962
references.citation: Int J Immunopharmacol. 1985;7(1):103-9
references.pmid: 3158616
references.citation: J Mol Biol. 2010 Apr 9;397(4):917-31
references.pmid: 20156456
references.citation: APMIS. 1997 Aug;105(8):643-9
references.pmid: 9298104
references.citation: J Nutr. 2007 Mar;137(3 Suppl 2):798S-802S
references.pmid: 17311978
references.citation: Can J Microbiol. 2009 Jun;55(6):648-56
references.pmid: 19767834
references.citation: Eur Urol. 1995;27(2):104-9
references.pmid: 7744150
references.citation: Med Microbiol Immunol. 1999 Dec;188(3):111-6
references.pmid: 10776840
references.citation: Transplant Proc. 2007 Dec;39(10):3219-21
references.pmid: 18089357
references.citation: Immunology. 2009 Sep;128(1 Suppl):e170-80
references.pmid: 19740306
references.citation: Int J Immunopathol Pharmacol. 2011 Jan-Mar;24(1 Suppl):45S-50S
references.pmid: 21329565
references.citation: J Natl Cancer Inst. 2008 Mar 5;100(5):359-72
references.pmid: 18314475
references.citation: Life Sci. 1998;63(8):635-44
references.pmid: 9718093
references.citation: Hum Cell. 2011 Mar;24(1):9-12
references.pmid: 21547690
references.citation: Toxicon. 2011 Aug;58(2):179-86
references.pmid: 21658402
references.citation: Planta Med. 2008 Nov;74(14):1693-700
references.pmid: 18951337
references.citation: Nutr Cancer. 2003;46(2):197-201
references.pmid: 14690796
references.citation: Lett Appl Microbiol. 2006 May;42(5):452-8
references.pmid: 16620202
references.citation: Int J Immunopharmacol. 1994 Jan;16(1):29-36
references.pmid: 8150553
references.citation: Mol Cancer Ther. 2007 Oct;6(10):2696-707
references.pmid: 17938263
references.citation: Proc Natl Acad Sci U S A. 2004 Sep 28;101(39):14228-33
references.pmid: 15381773
references.citation: Cancer Invest. 2009 Feb;27(2):163-70
references.pmid: 19235588
references.citation: Cancer Lett. 2010 Feb 1;288(1):1-9
references.pmid: 19523754