Dihydromyricetin inhibits migration and invasion of hepatoma cells through regulation of MMP-9 expression.

Abstract

AIM:

To investigate the effects of dihydromyricetin (DHM) on the migration and invasion of human hepatic cancer cells.

METHODS:

The hepatoma cell lines SK-Hep-1 and MHCC97L were used in this study. The cells were cultured in RPIM-1640 medium supplemented with 10% fetal bovine serum at 37 °C in a humidified 5% CO2 incubator. DHM was dissolved in dimethyl sulfoxide and diluted to various concentrations in medium before applying to cells. MTT assays were performed to measure the viability of the cells after DHM treatment. Wound healing and Boyden transwell assays were used to assess cancer cell motility. The invasive capacity of cancer cells was measured using Matrigel-coated transwell chambers. Matrix metalloproteinase (MMP)-2/9 activity was examined by fluorescence analysis. Western blot was carried out to analyze the expression of MMP-2, MMP-9, p-38, JNK, ERK1/2 and PKC-δ proteins. All data were analyzed by Student's t tests in GraphPad prism 5.0 software and are presented as mean ± SD.

RESULTS:

DHM was found to strongly inhibit the migration of the hepatoma cell lines SK-Hep-1 (without DHM, 24 h: 120 ± 8 μmol/L vs 100 μmol/L DHM, 24 h: 65 ± 10 μmol/L, P < 0.001) and MHCC97L (without DHM, 24 h: 126 ± 7 μmol/L vs 100 μmol/L DHM, 24 h: 74 ± 6 μmol/L, P < 0.001). The invasive capacity of the cells was reduced by DHM treatment (SK-Hep-1 cells without DHM, 24 h: 67 ± 4 μmol/L vs 100 μmol/L DHM, 24 h: 9 ± 3 μmol/L, P < 0.001; MHCC97L cells without DHM, 24 h: 117 ± 8 μmol/L vs 100 μmol/L DHM, 24 h: 45 ± 2 μmol/L, P < 0.001). MMP2/9 activity was also inhibited by DHM exposure (SK-Hep-1 cells without DHM, 24 h: 600 ± 26 μmol/L vs 100 μmol/L DHM, 24 h: 100 ± 6 μmol/L, P < 0.001; MHCC97L cells without DHM, 24 h: 504 ± 32 μmol/L vs 100 μmol/L DHM 24 h: 156 ± 10 μmol/L, P < 0.001). Western blot analysis showed that DHM decreased the expression level of MMP-9 but had little effect on MMP-2. Further investigation indicated that DHM markedly reduced the phosphorylation levels of p38, ERK1/2 and JNK in a concentration-dependent manner but had no impact on the total protein levels. In addition, PKC-δ protein, a key protein in the regulation of MMP family protein expression, was up-regulated with DHM treatment.

CONCLUSION:

These findings demonstrate that DHM inhibits the migration and invasion of hepatoma cells and may serve as a potential candidate agent for the prevention of HCC metastasis.

• Authors: Qing-Yu Zhang, Ran Li, Guo-Fang Zeng, Bin Liu, Jie Liu, Yang Shu, Zhong-Kao Liu, Zhi-Dong Qiu, Dong-Jun Wang, Hui-Lai Miao, Ming-Yi Li, Run-Zhi Zhu
• Journal: World journal of gastroenterology (World J Gastroenterol) Vol. 20 Issue 29 Pg. 10082-93 (Aug 07 2014) ISSN: 2219-2840 [Electronic] United States
• PMID: 25110435 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents, Phytogenic
• Flavonols
• Matrix Metalloproteinase Inhibitors
• PRKCD protein, human
• Protein Kinase C-delta
• Mitogen-Activated Protein Kinases
• MMP2 protein, human
• Matrix Metalloproteinase 2
• MMP9 protein, human
• Matrix Metalloproteinase 9
• dihydromyricetin

Topics

• Antineoplastic Agents, Phytogenic (pharmacology)
• Carcinoma, Hepatocellular (enzymology, pathology)
• Cell Adhesion (drug effects)
• Cell Line, Tumor
• Cell Movement (drug effects)
• Cell Survival (drug effects)
• Dose-Response Relationship, Drug
• Down-Regulation
• Flavonols (pharmacology)
• Gene Expression Regulation, Enzymologic (drug effects)
• Gene Expression Regulation, Neoplastic (drug effects)
• Humans
• Liver Neoplasms (enzymology, pathology)
• Matrix Metalloproteinase 2 (metabolism)
• Matrix Metalloproteinase 9 (metabolism)
• Matrix Metalloproteinase Inhibitors (pharmacology)
• Mitogen-Activated Protein Kinases (metabolism)
• Neoplasm Invasiveness
• Phosphorylation
• Protein Kinase C-delta (metabolism)
• Signal Transduction (drug effects)
• Time Factors