Effect of Silymarin on Expression of micro-RNA-21 and Matrix Metalloproteinase (MMP) 2 and 9 and Tissue Inhibitors of Matrix Metalloproteinase (TIMP) 1 and 2 in Hepatocellular Carcinoma Cell Line (HepG2).
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Abstract
Background: Silymarin is a flavonolignan that has various medicinal properties such as liver protection, antioxidant, anti-inflammatory, anti-cancer and heart protection activities. The aim of this study was to investigate the effect of silymarin on the expression level of mir-21, matrix metalloproteinase(MMP), and their tissue inhibitors (TIMPs) in liver cancer HepG2 cell line.
Methods: An in-vitro experimental study was conducted on the human HepG2 cells prepared from Pasteur Institute, Tehran, Iran. Four concentrations of 0 (control), 50, 100, and 150 µM of silymarin were considered as the study groups according to the MTT assay. Gene expression study was performed using real-time PCR. The studied genes were mir-21, MMP-2, MMP-9, TIMP-1 and TIMP-2. In addition, some apoptosis-related genes including BAX, BCL2 and Caspase3 (CAS3) were investigated. GAPDH was used as an internal control. Relative expression was calculated by REST program using t-test on the logarithm of expression considering a significance level of 0.05.
Results: The significant up-regulations consisted of TIMP genes for doses 100 µM and 150 µM, and the apoptosis activating genes CAS3 and BAX (P < 0.05). The significant down-regulations consisted of MMP-9 in all concentrations, MMP-2 in concentration 100 µM, and the apoptosis inhibitory gene BCL2 in concentrations 50 µM and 100 µM (P < 0.05). In addition, mir-21 as an oncogenic micro-RNA showed significant down-regulation for all doses (P < 0.05). All the comparisons were with the control group.
Conclusion: The present study showed that silymarin could affect the HepG2 cell line at the gene expression level via increasing apoptosis and changing the expression of MMP-2, MMP-9, TIMP-1, TIMP-2 and mir-21. These findings were in line with each other and in favor of suppression of tumoral activity in this cell line.
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