| Glioma is the most common malignant tumor in the central nervous system.The main clinical treatment is surgery,radiotherapy and chemotherapy.Although these methods can play a certain role,they are prone to relapse soon after treatment.There are toxic side effects and the quality of life of patients decline,so the exploration of effective and less toxic drugs has become the focus of the treatment of glioma.Myricetin is a natural flavonoid,mainly found in daily fruits and vegetables,tea,red wine and so on.Studies have shown that myricetin has antioxidant,anti-inflammatory,anti-tumor effects,and in vivo and in vitro can be used to prevent skin cancer.Colon cancer and other tumors play an antitumor role.Therefore,our team used myricetin to study the treatment of human glioma and explore its mechanism.In this experiment,the effects of myricetin on the activity,invasion and migration of human glioma cells and the ability of HUVEC tube formation were detected by CCK-8 test,scratch test and HUVEC tube forming experiment in vitro.Secondly,the effect of myricetin on MMP-2?MMP-9?MMP-14?MMP-26?HIF-1?and VEGF protein was detected by western blot assay.Finally,human glioma cell line U87 MG was subcutaneously inoculated on the back of male nude mice to establish a model to study the effect of myricetin on the growth of human glioma in vivo.1.The effect of myricetin on the proliferation of human glioma cellsThe effect of myricetin on the activity of human glioma cells by CCK-8 method was used to detect the effect of different concentrations of myricetin on the activity of U87 MG in human glioma cells.The results showed that the inhibitory effect of cell activity increased with the increase of myricetin concentration,when the concentration increased to 80?M,myricetin to human brain compared with the control group.The inhibitory effect of glioma cell activity was statistically significant(P<0.05).The results showed that myricetin could inhibit the activity of human glioma cells in a concentration dependent manner.2.Effects of myricetin on migration,invasion and angiogenesis of human gliomacells2.1 The effects of different concentrations of myricetin on the migration of human glioma cells were detected by scratch test.The results showed that the migration ability of human glioma cells decreased with the increase of myricetin concentration,which was significantly lower than that of the control group(P<0.05).2.2 The effects of different concentrations of myricetin on the invasiveness of human glioma cells were detected by transwell chamber assay.The results showed that with the increase of myricetin concentration,the number of cells passing through the matrix into the lower chamber gradually decreased,compared with the control group,there was statistical significance(P<0.05).2.3 The influence of myricetin on HUVEC tube forming ability was tested by huvec on matrigel.The results showed that with the increase of myricetin concentration,myricetin could inhibit the formation of HUVEC tube,which had statistical significance compared with the control group(P<0.05).2.4 The expression of MMP-2,MMP-9,MMP-14,MMP-26,HIF-1?,VEGF protein was detected by western blot.The results showed that the expression of MMP-2,MMP-9,MMP-14,MMP-26,HIF-1?,VEGF protein decreased with the increase of myricetin concentration in human glioma cells.Compared with the control group,there was significant difference(P<0.05).1.The effect of myricetin on the growth of human gliomaThe nude mice were divided into five groups: Solvent control group,temozolidomide positive control group,20 mg/kg myricetin group,40 mg/kg myricetin group,20 mg/kg myricetin combined with temozolidomide group.On the10 th day after inoculation,tumor bearing was formed and the body weight and tumor volume of nude mice were monitored daily by continuous intraperitoneal injection for10 days at a dose of 200?l per mouse.The results showed that the weight and volume of tumor in the experimental group were significantly different from those in the control group after administration(P<0.05).The combined administration group was statistically significant(P<0.05)with respect to the 20 mg/kg administered group alone;The expression of vascular endothelial marker CD31 in each group wasdetected by immunohistochemistry.The expression of CD31 was more in the solvent control group than that in the other four groups.The results showed that the positive expression of CD31 in the other four groups was lower than that in the solvent control group.The results showed that myricetin could inhibit the angiogenesis of the tumor,compared with the control group,there was statistical significance(P<0.05).Based on the above results,we can draw the following conclusions: myricetin inhibits the migration,invasion of human glioma cells in vitro,and myricetin of HUVEC inhibits the growth of human glioma in vivo. |
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