Growth Inhibition of Hepatocellular Carcinoma Huh7 Cells by Lactobacillus casei Extract.
10.3349/ymj.2013.54.5.1186
- Author:
Dae Jong HAN
1
;
Jong Bin KIM
;
Seo Young PARK
;
Man Gil YANG
;
Hyuncheol KIM
Author Information
1. Biomedical Research Institute, Seoul National University Hospital, Seoul, Korea.
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Lactobacillus casei;
liver cancer;
proliferation;
side population;
cancer stem-like cell
- MeSH:
Apoptosis/drug effects;
Carcinoma, Hepatocellular/*pathology;
Caspase 8/metabolism;
Caspase 9/metabolism;
Cell Cycle Checkpoints/drug effects;
Cell Extracts/*pharmacology;
Cell Line, Tumor;
Cell Proliferation/drug effects;
Cyclin-Dependent Kinase Inhibitor p21/metabolism;
Cyclin-Dependent Kinase Inhibitor p27/metabolism;
Cytostatic Agents/*pharmacology;
Flow Cytometry;
Gene Expression Regulation, Neoplastic/drug effects;
Humans;
Lactobacillus casei/*chemistry;
Liver Neoplasms/*pathology;
Proto-Oncogene Proteins c-bcl-2/metabolism;
RNA, Messenger/metabolism;
Receptors, Tumor Necrosis Factor, Member 25/metabolism;
Receptors, Tumor Necrosis Factor, Type I/metabolism;
bcl-Associated Death Protein/metabolism
- From:Yonsei Medical Journal
2013;54(5):1186-1193
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: 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. MATERIALS AND 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-alpha 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. CONCLUSION: L. casei extract exerts a potent anti-tumor effect on the viability of liver cancer cells, although not on cancer stem-like cells.
