Abstract

 
 
 
 Purpose: To study the impact of matrine on cell cycle and apoptotic changes in hepatoma cells, and the mechanism involved.
 Methods: Human hepatoma cell line HepG2 was treated with different concentrations of matrine. The blank control cells were maintained in 1640 medium only. The influence of matrine on proliferative ability was determined with 3-(4,5)-dimethylthiahiazo (-z-y1)-3,5-di- phenytetrazoliumromide (MTT) method. Flow cytometry was used to determine its effect on cell cycle and apoptosis; RT-PCR (reverse transcription-polymerase chain reaction) was applied to assay the mRNA expressions of miR-122, cyclin G1 (CG1), livin and survivin mRNA, while the protein expressions of CG1, livin and surviving were assayed by Western blotting.
 Results: Matrine time- and dose-dependently suppressed the proliferative capacity of the cells. At a concentration of 0.5 mg/mL, matrine had no significant effect on the cell cycle. However, 1.0 mg/mL matrine blocked the cell cycle in G1 phase, while 1.5 mg/mL matrine blocked HepG2 cells in G2/M phase (p < 0.05). Moreover, matrine induced apoptosis in HepG2 cells, and markedly downregulated the expressions of miR-122 concentration- time-reliantly (p < 0.05). In addition, matrine markedly and concentration-dependently reduced mRNA and protein expression levels of CG1, livin and survivin, with the strongest inhibitory effect at a level of 1.5 mg/mL.
 Conclusion: Matrine induces cell cycle block and apoptotic changes in hepatoma cells through a mechanism related to regulation of the CG1/livin/survivin signal axis mediated by miR-122. Matrine may be a potential treatment for liver cancer. However, clinical trials are needed to confirm this potential.
 
 
 
Highlights
Liver cancer is a frequently diagnosed primary carcinoma, accounting for about 90 % of all cases
The pathogenesis of liver cancer is associated with multiple factors and multiple gene interactions
Studies have shown that TCMs induce apoptosis by arresting the cell cycle at some phases, and studies have demonstrated that telomerase plays an important role in cell immortalization and malignancy [7,8]
Summary
Liver cancer is a frequently diagnosed primary carcinoma, accounting for about 90 % of all cases. Frozen HepG2 cells were taken out and thawed in a 37 oC water bath. Three groups of cells at logarithmic phase were set up. They were exposed to matrine at doses of 0.5, 1.0, 1.5 mg/mL. Cells in the blank control received 1640 medium only. Logarithmic growth-phase cells were seeded in 96-well plates after removal of culture medium and rinsing with PBS. They received either matrine at doses of 0.5, 1.0, 1.5 mg/mL, or 1640 medium only, followed by culturing at 37 °C and 5 % CO2 for 48 h. All statistical analyses were carried out with SPSS20.0 software package
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