Reaserch On The Effect Of MiR-25 On Regulatory T Cell Function In Advanced Primary Hepatic Cancer

Objective:This study was to determine the difference in cell frequency of peripheral blood Treg between Primary hepatic carcinoma(PHC)patients and healthy controls,and to analyze whether Treg has abnormal proliferation in PHC.Explore the target genes and mechanism of mi R-25,and analyze whether there is a signal pathway between mi R-25 and Treg,which provides new ideas and ways for the prevention and treatment of PHC.The aim of this study was to detect the expression level of mi R-25 in peripheral blood serum of patients with PHC,and to study the interaction between mi R-25 and Treg,and provide new ideas and ways for the prevention and treatment of PHC.Methods:1.To collected complete clinical data of 20 patients of PHC in Subei People's Hospital liver and spleen medicine and liver and spleen surgery from January 2018 to October 2018,and their serum divided into experimental group.Collected 20 healthy volunteers were healthy controls.The serum of 20 healthy volunteers was collected as a healthy control group.2.To detect the frequency of peripheral blood Treg cells in experimental and healthy controls by Flow Cytometry.Statistical analysis the difference in peripheral blood Treg cell frequency between the experimental group and the healthy control group.3.To predict the target genes of mi R-25,and to analyze whether the predicted target genes have targeted regulation by Bioinformatics methods.Analyze the possible signaling pathways between mi R-25 and Treg by checking reference literature.4.To detection the mi R-25 expression in peripheral blood serum of experimental group and healthy control group by Polymerase Chain Reaction(PCR).Results:1.The difference was statistically significant(***P<0.001),and number of Treg cells in 1 million peripheral blood cells of patients with PHC was higher than that of healthy controls group?2.The experimental group's cell frequency of Treg cells was(5.42±6.85)%,and the healthy control group's cell frequency of Treg cells was(4.04±6.85)%,and the difference between two group was significant(***P<0.001).3.Bioinformatics suggests that mi R-25 has a conserved binding site to the 3' URT of sphingosine 1-phosphate receptor 1(S1P1)in a variety of organisms.Studies have shown that S1P1 acts as a unique receptor system that negatively regulates the function of Treg cells.Summarizing the literature,S1P1 inhibits thymus generation by activating the downstream Akt-m TOR(S1P1-PI3K-Akt-m TOR)signaling pathway,providing a negative signal for the immunosuppressive effect of Treg cells.4.The expression of mi R-25 in peripheral blood of patients with PHC was higher than that of healthy controls group(P=0.087),and the difference was statistically significant(**P<0.01).Conclusions:The expression level of mi R-25 in peripheral blood of patients with advanced PHC is higher than that of healthy controls.Targeting regulation of S1P1 results in low expression of this molecule.S1P1 inhibits the thymic generation by activating the S1P1-PI3K-Akt-m TOR signaling pathway,inhibits the activity of Treg cells,and affects the immune tolerance and immune escape of tumor cells.Therefore,the increase of mi R-25 level leads to the negative regulation of S1P1 on the growth and development of Treg cells,which increases the expression of Treg cells in PHC patients and exerts immunomodulatory effects on PHC.Therefore,mi R-25 will be a potential target for the prevention and treatment of PHC and is expected to become a new biomarker for PHC diagnosis.