Abstract
Purpose: To determine the inhibitory effect of glycyrrhizin (GLA) on cell invasion and metastasis in mammary carcinoma cells, and the mechanisms of actions involved.Methods: The effect of GLA at different concentrations on proliferation of breast cancer MDA-MB-231 and BT549 cells was assayed by MTT method. Transwell assay was used to determine the effect of GLA at different concentrations on invasiveness and metastasis of breast cancer MDA-MB-231 and BT549 cells. The influence of LGA on expressions of microRNA-200c and miR-200c was assayed by reverse transcriptase-polymerase chain reaction (RT-PCR).Results: There was no statistically significant difference in cell proliferation amongst cells treated with 5 and 20 μM GLA and untreated breast cancer cells. However, the proliferation of cells treated with 40 μM GLA was significantly reduced (p < 0.05). In the cell invasion and migration experiments, cell population transferred to the base of Transwell chamber in the two cell lines treated with GLA was markedly decreased, relative to cells without GLA treatment, while the number of cells decreased with increase in GLA concentration (p < 0.05). Results from image-pro-plus analysis revealed that the population of cells quantitatively crossing the Transwell compartment membrane decreased with increase in GLA concentration (p < 0.05). The expression of e-cadherin was increased by GLA treatment in a concentration-dependent manner. Moreover, GLA treatment led to significant changes in amounts of miR-200s a, b and c, with changes in miR-200c being the most significant (p < 0.05).Conclusion: GLA suppresses the invasiveness and metastasis of breast cancer MDA-MB-231 and BT549 cells via upregulation of the expressions of miR-200c and e-cadherin. These findings provide a theoretical basis for the development of new breast cancer drugs.
 Keywords: Glycyrrhiza, GLA, miR-200c, E-cadherin, Inhibition, Breast cancer cells, Invasion, Metastasis
Highlights
Mammary carcinoma is among the most frequently diagnosed malignancies among women, and a major reason for female mortality in the world
The purpose of this study was to determine the relationship between GLA-induced inhibition of breast cancer cell invasion and metastasis, and up-regulation of e-cadherin expression through miR-200c
Effect of different GLA treatments on breast cancer cell invasion and metastasis In the cell invasion and migration experiments, the population of cells transferred to the base of the Transwell chamber in the two cell lines treated with GLA was markedly decreased, relative to cells without GLA treatment, and the number of cells decreased with increase in GLA concentration
Summary
Mammary carcinoma is among the most frequently diagnosed malignancies among women, and a major reason for female mortality in the world. Two days after inoculation and culture in a 96-well plate, 1ml MEM medium was transferred to a 1.5 mL centrifuge tube, and the required treatment drugs and transfection reagent were added. 500 μL of methanol was added to 24-well plate This was followed with washing twice with PBS. The original solution of Matrigel was diluted with serum-free and double antibody-free medium in a volume ratio of 1:8, and 100 μL was added to the chamber, which was dried in a cell incubator for 2 h. The cells were fixed with methanol (500 μL) for 25 min 500 μL of crystal violet was put in the well plate in the invasion chamber for 25 min, after which it was PBS-rinsed two times.
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