| Objective:MiRNAs have been shown to be involved in human placental production.During embryonic implantation,mi RNAs are involved in inflammatory gene-related expression and are involved in maternal immune tolerance.Some key molecules in miRNA production processes such as DICER,DROSHA and RAN regulate miRNA production,Thereby regulating the expression of genes and proteins within reproductive process,It has beenconfirmed that miRNAs are closely related to human reproduction,Recurrent abortion is a complex multifactorial disease,of which 50%to 70%could be classified asunexplained recurrent spontaneous abortion(URSA),whichmay be potential genetic defects such as genetic polymorphism.his study aims to explore the association between the polymorphisms of mi RNA biogenesis related gene DICER,DROSHA,RAN and unexplained recurrent spontaneous abortion(RSA)in Chinese Women.Methods:From June 2013 to June 2016,Department of reproductive endocrinology and genetics and eugenics,West China Second Hospita,Sichuan University and the Department of Obstetrics and Gynecology in Chengdu Second people’s Hospital,we collected 217 unexplained recurrent spontaneous abortion patients.Patients were diagnosed with URSA who did a series of laboratory tests,including chromosome karyotype,thyroid function,reproductive immunity,sex hormones,B ultrasound,genital tract infection,TORCH,coagulation,liver and kidney function.And 390 healthy fertile women with a history of one or more times successful pregnancy outcome were recruited as control,The control group are coming from physical examination and prepregnancy check for women of childbearing age in the same period.We performed a case-control study to analyze polymorphism of miRNA machinery genes,including DICER rs3742330,DROSHA rs10719,RAN rs14035,by polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP)assay.Electrophoretic gel was used to genotype,and combined with gene sequencing to verify it,In this study,we used the?~2 test to compare the allele and genotype frequencies in two groups,and the univariate logistic regression was used to calculate the most significant genetic pattern of the risk allele.According to the genetic model,the multivariate logistic regression model was used to analyze the relationship between Genotype Combination Effect of the three SNPs and unexplained recurrent miscarriage.Results:The genotype distribution of DICERrs3742330,DROSHArs10719 and RANrs14035 were consistent with Hardy-Weinberg’s law(P>0.05),respectively,in the case and control group.The result of PCR product sequence was consistent with that of restriction enzyme gel electrophoresis.The frequencies of genotypes and alleles of DICER rs3742330,DROSHA rs10719,and RAN rs14035 showed no difference in patients and control group(all P>0.05).univariate logistic regression showed that DICER3742330 tend to recessive(P=0.007),DROSHArs10719(P=0.088)and RANrs14035(P=0.031)tend to be dominant in the dominant pattern of risk alleles.According to the genetic model determined by each locus,We furtherjointly analyzed the polymorphisms of miRNA maturation-related genes which are DICER rs3742330,DROSHArs10719 and RAN rs14035,The results showed that DICER rs3742330/DROSHArs10719GG/TC+TT combinations were synergistically associat-ed with an increased URSA risk(OR=1.657,95%CI=1.006-2.731,P=0.047),which is risk factor.At the same time,with DROSHA10719 as a reference,Although DICER rs3742330/RANrs14035GG/CC combinations was significantly higher in the abortion group than in the control group,there was no Statistical significance.(OR=1.977,95%CI=0.956-4.087,P=0.066).Howere,DROSHA10719/RAN14035TT+TC/CC had no obvious correlation With URSA(OR=0.958,95%CI=0.679-1.353,P=0.808).Conclusions:In a single locus genotype analysis,we found no significant correlation between DICER rs3742330,DROSHA rs10719,RAN rs14035 and unexplained recurrent miscarriage,However,our study demonstrated the relationship between URSA development and combined genotype of DICER rs3742330/DROSHA rs10719 GG/TC+TT. |
