| Background:Colorectal cancer(CRC)is not only the third most common cancer in the world,but also the fourth most fatal cancer in the world.In 2020,the mortality rate caused by CRC will reach9.4%.The recurrence and metastasis of colorectal cancer is an important cause of death.Mi RNAs belong to a large class of non coding RNAs(nc RNAs).They are highly conserved non coding RNAs.Their angiogenesis is closely related to tumor recurrence and metastasis.Jiedu Yiqi Quyu prescription is a prescription summarized by the research group on the basis of basic experimental research and clinical research of colorectal cancer for many years.Its curative effect is positive,and it has a certain degree of anti recurrence and metastasis effect on colorectal cancer,but the specific mechanism is still unclear,which needs to be further studied.Objective:In this study,we started with miRNAs,used bioinformatics methods in the early stage,used various online databases and software according to the research needs,combined with literature research to explore the differential expression of miRNA in colorectal cancer,collected clinical samples to verify the expression of miR-133 a in colorectal cancer in the later stage,and explained the action mechanism of miR-133 a in colorectal cancer by cell culture intervention of traditional Chinese medicine serum To enrich the therapeutic mechanism of Tonifying Qi and removing toxin.Methods:1.Bioinformatics methods: the published colorectal cancer data sets were downloaded from the geo database in the big data environment,and the differentially expressed miRNAs in the data sets were analyzed by geo2 r software.2.Tissue specimen detection: 30 cases of primary colorectal cancer tissues and adjacent tissues diagnosed by pathology were selected,and no other non-surgical treatment was performed before operation.Real time quantitative PCR was used to detect the expression of target miRNA.Immunohistochemistry was used to detect the formation of MVD with CD31 positive expression.Pearson correlation analysis was used to explore the correlation between the expression of mir-133 a and MVD in colorectal cancer.3.In vitro experiment: the serum containing Jiedu Yiqi Quyu formula was prepared by half male and half female SD rats.Lovo-huvecs were cocultured with conventional methods.According to the dosage,the low,medium and high dosage groups were set up,including Jiedu Yiqi Quyu formula containing serum group and three control groups(positive control drug 5-FU group,blank control group and solvent control group).CCK-8 was used to detect the inhibition rate of different dosage of drug containing serum group and 5-FU on HUVEC cells;Transwell chamber was used to observe the inhibition rate of different dosage of drug containing serum group and 5-FU on HUVEC cells-Objective: To explore the mechanism of mir-133 a / VEGFA /VEGFR2 in the intervention of Jiedu Yiqi Quyu formula.Methods: HUVEC cells were divided into four groups: the inhibition of Fu on HUVEC cell migration;the inhibition of drug containing serum on HUVEC tube formation;the total protein of cells in the intervention of drug containing serum group and control group were extracted;the expression of mir-133 a was detected by q RT-PCR;the expression of angiogenesis related targets VEGFA and VEGFR2 were detected by Western blot.Results:1.Bioinformatics results showed that miR-133 a may be one of the differentially expressed miRNAs in colorectal cancer.2.The positive rate of MVD of CD31 in cancer tissue was 22.342 ± 5.392,and that of MVD in paracancerous tissue was 7.927 ± 2.050;the expression of mir-133 a in colorectal cancer tissue was lower than that in paracancerous tissue.3.CCK-8 results showed that the serum containing different doses of Jiedu Yiqi Quyu formula could inhibit the proliferation of HUVEC cells to a certain extent 3 days after administration,showing a positive correlation with time and dose.4.Transwell results showed that Jiedu Yiqi Quyu formula containing serum of different doses could inhibit HUVEC cell migration to a certain extent.5.The tube forming test showed that Jiedu Yiqi Quyu formula containing serum could inhibit the tube forming of HUVEC to a certain extent.6.Western blot results showed that: compared with the solvent control group and the blank control group,the expression of VEGFA and VEGFR2 protein in the cells intervened by the serum containing Jiedu Yiqi Quyu recipe decreased(P < 0.05),and the expression of mir133 a increased,suggesting that the serum containing Jiedu Yiqi Quyu recipe may inhibit the angiogenesis of colorectal cancer by regulating miR-133a/VEGFA/VEGFR2 signaling pathway.Conclusion:1.miR-133 a is down regulated in colorectal cancer,which may participate in the occurrence and development of colorectal cancer by intervening in angiogenesis.It is a new potential target for the treatment of colorectal cancer;2.Jiedu Yiqi Quyu herbs can inhibit the proliferation,migration and tube formation of HUVEC cells;3.Jiedu Yiqi Quyu herbs may increase the expression level of mir133 a and down regulate the expression of VEGFA and VEGFR2 in a dose-dependent manner... |
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