Theeffectand Mechanism Of Glycyrrhizic Acid On Human Colon Cancer Lovo Cells

Objective:To observe the effect of glycyrrhizic acid on the proliferation and migration of human colon cancer lovo cells and explore the possible mechanism.Methods:1.Human colon cancer lovo cells were divided into four groups.The control group,G1 group(glycyrrhizic acid100ug/L),G2 group(glycyrrhizic acid200ug/L)and G3 group(glycyrrhizic acid400ug/L).The control group only adds medium.2.CCK-8 was used to detect the effect of different concentrations of glycyrrhizic acid on the proliferation of lovo cells.Lovo cells were administered on 96-well culture plate.According to the above grouping,Lovo cells in 4 groups were placed at 37℃ 5% CO2 incubator for 24,48,72 hours,and added the CCK-8 solution respectively.Absorbance value was measured on the automatic enzyme-linked immune tester in 490 nm wavelength.The above operation repeats 3 times in each group,taking the average to calculate cell proliferation inhibition rate.Calculation formula:(absorbance value of glycyrrhizic acid experimental group-absorbance value of background group)/(absorbance value of the control group-absorbance value of background group)x100%.3.Transwell assay was used to determine the effects of differentconcentrations of glycyrrhizic acid on the migration of lovo cells.Lovo cell in 4 groups wereinoculated in the upper of transwellchamber,and the DMEM culture solution were putted into the lower room.Lovo cell in 4 groups were placed into 37℃ 5% CO2 incubator for cultivating 24 h,and then the crystal violet solution was putted.After being putted into incubator for 30 minutes cultivating,numbers ofLovo cellswere photographed by a microscope and counted.The experiment mentioned above was repeated three times.4.The expression of β-catenin protein and Cyclin D1 protein of Lovo cells in 4 groups were detected by western-blot after 24 hours.5.The expressions of β-catenin mRNA and Cyclin mRNA of Lovo cells in 4 groups were detected by RT-PCR after 24 hours.Results:1.Glycyrrhizic acid can inhibit the proliferation of Lovo cells.The higher concentration of glycyrrhizic acid,the longer it works,the more obvious in inhibition,showing a concentration-dependent manner.2.Glycyrrhizic acid can inhibit the migration of lovo cells.The higher the concentration of glycyrrhizic acid,the more obvious in inhibition,showing a concentration-dependent manner.3.Glycyrrhizic acid can inhibit the expression of Wnt signal pathway-related proteins,β-catenin,and its target protein Cyclin D1.The higher the concentration of glycyrrhizic acid,the more obvious in inhibition,showing a concentration-dependent manner.4.Glycyrrhilic acid can inhibit the expression of Wnt signaling pathway related genes,β-catenin mRNA and Cyclin D1 mRNA in Lovo cells.The higher the concentration of glycyrrhizic acid,the more obvious in inhibition,showing a concentration-dependent manner.Conclusion:1.Glycyrrhizic acid can inhibit the proliferation and migration of colon cancer Lovo cells.2.Glycyrrhizic acid can play a role by inhibiting the Wnt/ β-catenin signal pathway.