MiRNAs Are Studied In Embryo Implantation And Human Diseases

一、The function of MiR-320 aned miR-320 during embryo implantationIn recent years,the research about miRNA has become the hot spot in life science theory and application research.In theory,this paper studies miRNAs how to adjust the gene transcription expression patterns in cell differentiation,proliferation and apoptosis process,then the effects in the process of embryos implantation and post-implantation;In application,miRNA tissue specific expression patterns and the SNP change may be the new symbol in disease diagnosis and early warning.We studied the role of miRNA in embryos implantation and Alzheimer’s disease from miRNA aspects.Objective:To identify differential expression of two miRNAs in the rats uterus between pre-implantation period and implantation period.To research the expression and regulation of miR-143 and miR-320 in the process of embryo implantation in rats and in HUFCS cell.This study has the potential to provide new insights into the mechanisms of embryo implantation.Methods:1.Northern blot and in situ hybridization was utilized to analyse expression and distribution of miR-143 and miR-320 during peri-implantation period.2.MiR-143 and miR-320 expression was detected in uterus of pseudopregnancy,artificial decidualization,delayed-implantation and ovariectomized rats treated with estradiol-17β or progesterone by Northern blot and in situ hybridization.3.Over expression or knocking down miR-143 and miR-320 make the detection of HUFCS cells proliferation,apoptosis,migration,the influence of infiltration function by Edu,MTT,Tunel and Transwell.4.Changing cells miRNA level observe the mRNA and protein levels of the endogenous LIFR change by using of fluorescent element enzyme report system and western blot.Results:1.The expression level of miR-143 was higher during implantation and post-implantation in rats than during the pre-implantation periods.MiR-143 was specifically localized in maternal gland and luminal epithelia and stroma and decidua.MiR-320 was differentially expressed in the rat uterus between pre-implantation period and implantation period.MiR-320 was specifically localized in the s maternal gland.2.The expression of miR-143 and miR-320 was not significantly different in the pseudopregnant uterus.The expression of miR-143 can be increased,but the expression of miR-320 can be inhibited in uterus delayed-implantation.Artificial decidualization can promote the expression of miR-143,but is not significantly different to the miR-320.Treatment with estradiol-17β and progesterone didn’t significantly change miR-143 expression.Treatment with progesterone upregulated miR-320 expression.3.Over expression miR-143 make the HUFCS the ability of the infiltrating stronger(P<0.05),transfer ability no changed,the proliferation capacity weaker(P<0.05),the apoptosis ability no changed.Over expression miR-320 make the HUFCS the ability of the infiltrating stronger(P<0.05),transfer ability no changed,the proliferation capacity stronger(P<0.05),the apoptosis ability no changed.4.Dual-luciferase Activity Assay showed that the luciferase activity of co-transfection of miR-143 and miR-320 with PGL3-LIFR 3’-UTR were reduced.It suggests that miR-143 and miR-320 can interact with 3’-untranslated region(UTR)of LIFR.But overexpression or knocking out miR-143 can affect the luciferase activity,and overexpression of the miR-143 can inhibit the expression of the LIFR protein,the miR-320 has no significantly difference to the expression of the LIFR protein.It suggests that the miR-143 can regulate the expression of the LIFR,but the miR-320 can’t regulate the expression of the LIFR.Conclusion:1.The increased expression level of miR-143 was mainly caused by active blastocysts and decidualization in embryo implantation in the early of pregnancy in rats.MiR-143 may play an important role during embroy implantation in the rats and miR-143 can promote the embryo implantation.Mir-143 can regulate the ability of the infiltration and the proliferation in HUFCS.And the effects may be realized by adjusting the target gene LIFR.2.The decreased expression level of miR-320 was mainly caused by active blastocysts in embryo implantation in the early of pregnancy in rats.Steroid hormones,progesterone increased miR-320 expression.The knocking out of the miR-320 can promote the embryo implantation.Mir-320 can regulate the ability of the infiltration and the proliferation in HUFCS.And the effects may be realized by other way of adjusting the target gene LIFR.二、The correlation research between one SNP site in pri-miR-124 and Alzheimer’s disease in a Mongolian populationObjective:We detected the effect of one SNPs of miR-124 on miRNA biogenesis and analyzed the relationship between SNP rs531564 and AD.This study may provide new insights into the functions of miR-124 in the occurrence of nervous system diseases.Methods:1.PCR,genotypes analysis Sequencing,genetic Case-Control Association Study will show the mutation and the SNP.2.Secondary structure of the single nucleotide polymorphisms(SNPs)in miR-124 sequence will be showed by Bioinformatics prediction.3.Northern blot and Real-time PCR analysis will show the change of mature miR-124.Results:1.A common G/C polymorphism designated rs531564 was found in the pri-miR-124.2.The G allele changed the formation of a ring-shaped structure in the predicted secondary structure of the pri-miRNA for miR-124-1.3.The amount of mature miR-124 from the C/G heterozygosityof rs531564 was increased compared with the CC or GG homozygosity of rs531564.The expression of mature miR-124 from GG homozygosity was also higher than that from CC homozygosity(P<0.05).Conclusion:The present study is the first to evaluate the relationship between miR-124 and AD in the Mongolian population.SNP rs531564 of miR-124 may not represent a risk factor in the development of AD among Mongolian population.