Resveratrol Inhibits Angiogenesis In Canine Mammary Tumor

Mammary tumor is a tumorous disease that harms dog health seriously and cause organ failure through organ damage such as malignant proliferation,metastasis,and invasion.At this stage,most of the dog mammary tumors are treated with surgery and radiochemotherapy.However,side effects are accompanied and the prognosis easily recurs after the treatment.Resveratrol,as an extract of traditional Chinese medicine,Professor Jang M has reported in Nature in 1997 to discuss its anti-tumor effects.In recent years,resveratrol has gradually become a popular target for anti-tumor mechanism research.This study investigated whether resveratrol can influence the mechanism of angiogenesis and Notch signaling pathway in canine mammary tumor cells.CHMm,a canine breast tumor cell line with the same logarithmic growth phase,was selected and cells were treated with 10 ?M,20 ?M,40 ?M,80 ?M,and 160 ?M Res respectively.Then the cells of each group were collected for CCK-8 cell activity detection;Elisa was used to detect the levels of angiogenic factor(VEGF),matrix metalloproteinase(MMP-14)and hypoxiainducible factor(HIF-1?);real-time fluorescence quantitative PCR method The relative expression levels of Notch1,Jagged1,Dll4,and Hes-5 genes in tumor cells were detected;the expression levels of Notch1,Jagged1,Dll4,and Hes-5 proteins were detected by Western blot.1.CCK-8 cytotoxicity test results found that the same effect time,compared with the blank control group,the concentration of Res plus drug group on the inhibition of CHMm cells have a significant effect(p <0.01).At the same concentration of Res,the effect of Res on cell inhibition was significantly different(p<0.01)compared with the culture for 48 h.Res inhibition of CHMm cells showed a significant time and concentration dependence.2.The results of VEGF test showed that compared with the blank control group,the difference was not significant(p>0.05)except for the 10 ?M concentration of the drug-added group,and the effect of the decrease of VEGF content was extremely significant in the other groups with the increase of Res concentration(p<0.01).Therefore,the low concentration of Res had a weaker effect on the secretion of VEGF in canine mammary tumor cells.The medium and high concentration of Res had a significant inhibitory effect on the secretion of VEGF in a dosedependent manner.3.The results of HIF-1? showed that compared with the blank control group,the HIF-1?content decreased significantly with increasing Res concentration(p<0.01).Therefore,DHA significantly inhibited the secretion of HIF-1? in canine mammary tumor cells and the inhibitory effect was enhanced with the increase of Res concentration in a dose-dependent manner.4.MMP-14 results showed that compared with the blank control group,the difference of 10?M,20 ?M,and 40 ?M concentrations of Res was not significant(p>0.05),while the difference between 80 ?M and 160 ?M concentrations was extremely significant(p<0.01).Therefore,low and medium concentrations of Res had no significant effect on the secretion of MMP-14 fromcanine mammary tumor cells.Only high concentrations of Res produced significant inhibitory effects on the secretion of MMP-14.5.Notch-related mRNA expression results showed that,compared with the blank control group,Notch-related mRNA expression results showed that the mRNA expression levels of Notch1,Jagged1,Hes-5 and Dll4 were significantly down-regulated(0.01<p)in each concentration of Res administration group.(p<0.05),the target gene Hes-5,Dll4 extremely significant(p <0.01).6.The detection results of Notch-associated factor protein expression showed that,compared with the blank control group,the expression levels of Notch1,Dll4 and Hes-5 of the target protein at each concentration of Res administration were significantly down-regulated,but the Jagged groups were the same.The down-regulation of the amount of protein expression was not significant.Resveratrol has inhibitory effects on canine mammary tumor cells,and regulates the angiogenesis of inhibited cells through the expression of Notch pathway-related factor genes and proteins.