The Inhibition And Mechanisms Of Blueberries Anthocyanidin On Melanomas In C57BL/6 Mice

As one of the most aggressive malignancies in all the cancer types,Melanoma is highly invasive and easily metastatic features.However,there still is no drug found specificlly for melanoma.Hence it is of impendence to develop new therapeutic approaches for the malignancy.One principle method is to find active components from natural products.It has been testified that anthocyanin in grape collard and purple brussels sprout can inhibit melanoma in some degree.It is well known that blueberry has excellent nutritional value because of rich in anthocyanin.In this work,we paid attention on its inhibitory effect and mechanism for melanoma through urine metabolomic strategy and molecular biology methods.The work contianed four contents as listed below.1.At first,CCK-8 assay and acridine orange/ethidium bromide(AO/EB)staining were used to investigate whether blueberry anthocyanin had the inhibitory effect to model B16 cell of melanoma.The results showed blueberry anthocyanin can inhibit B16 cell growth and change the morphololy of it in some degree just as those of grape collards and purple brussels sprouts.It was concentration-and time-dependant.2.The unrine metabolomic strategy was used to compare the difference among the control,tumor model and given drug group via UPLC-TOF MS platform.Two potential biomakers were screened including nicotinic acid mononucleotide and 5-S-CD.It indicated that the inhibitory effect of blueberry anthocyanin for melanoma was achieved by inhabiting synthesis ncotinic acid mononucleotide and 5-S-CD.3.Molecular biology methods were used to investigate the possible mechanism of blueberry anthocyanin for melanoma.It was found that angiogenic protein vascular endothelial growth factor(VEGF)and two matrix metalloproteinases of MMP-2,MMP-9 expressed in tumor tissue significantly decreased after given drugs.Then the activity of MMP-2 and MMP-9 were investigated using a gelatin zymography assay.Compared with the tumor model,line brightness and the integral light density value reduced after treated with blueberry anthocyanin.All evidence demonstrated that blueberries anthocyanin inhibit activities of both MMP-2 and MMP-9,then reduce tumor invasion,metastasis.4.Moreover,inter-individual variability of the metabolic urinary profile was investigated between healthy populations of Uygur and Han,the largest population in Xinjiang,including race,age and gender.Totally 192 urine samples(96 Uygur and 96 Han)were collected and analyzed based on a gas chromatography-mass spectrometer platform.The data by orthogonal signal correction filtered partial least-squares discriminant analysis were show significant differences between urine of Uygur and Han.Totally eight metabolites are identified contributed to the discrimination between Han and Uygur,including phenylacetylglutamine,myoinositol,d-galactose,ribonolactone,octadecanoic acid,galactitol,threonic acid and succinic acid.Among those,three metabolites are being used as biomarkers in clinic.It suggests that the race effect should be critically considered prior to make diagnostic result in multi-race coexisted areas to decrease the false positive rate.Moreover,the results show that the age-period and the gender also affect the urinary metabolomics profiles.