Effect Of Folate,hnRNA E2 And Late Genes Of HPV16 And Their Interaction In Cervical Intraepithelial Neoplasia

Objective:Persistent infection of high risk HPV(HR-HPV)16 is the main cause of cervical cancer.The integration between HPV16 DNA and host genes is the key to cervical carcinogenesis.Late genes L1/L2,not only can be used as indicators of HPV infection,but also can be used as the symbol of the integration of virus and host chromosome.heterogeneous nuclear ribonucleoprotein E2(hnRNP E2)take part in gene transcription and translate regulation.Late genes contain multiple locsi and structures combined with RNA binding protein.The KH structure domain of hnRNP E2 provides the unique molecular structure which can combine with HPV16 late genes.Folate deficiency leads to the obstacle of DNA synthesis and methylation.Former research shows that folate deficiency may lead to the combination between hnRNP E1 and HPV16 RNA,decrease the expression of viral capsid protein and promote HPV16 DNA integration into host DNA.hnRNP E2 and hnRNP E1 belong to the same kind of RNA binding protein,and they have simliar structure.There is no paper about the interaction among folate,hnRNP E2 and late proteins of HPV16 in cervical intraepithelial neoplasia.The purpose of this study was to analyze the level of folic acid,hnRNP E2 and late proteins of HPV16 in different cervical intraepithelial neoplasia,and explore the role and interaction effect among folate,hnRNP E2 and late proteins of HPV16 in cervical intraepithelial neoplasia.Then we maybe provide evidence for the etiology and prevention of cervical intraepithelial neoplasia.Methods:The samples of this study were obtained from the cohort study of community cervicallesions in Jiexiu City of Shanxi Province from June 2014 to September 2014.The participants were new cases diagnosed by TCT,colposcopy and histology,including 61 women with normal cervical(NC),59 patients with low-grade cervical intraepithelial neoplasia(CIN ?),60 patients with high-grade cervical intraepithelial neoplasia(CIN?/?).All cases signed informed consent in this study.The general demographic characteristics,living habits,reproductive status,past medical history and other data were collected through structured questionnaire.Levels of serum folic acid were determined by chemiluminescence method.HPV infection was detected by diversion hybridization.Western blotting were used to detect the levels of the expression of hnRNP E2,HPV16 L1 and L2 respectively.c2 test,Logistic regression and Kruskal-Wallis H test were performed by SPSS19.0 software.The interaction effects between two factors were analyzed by additive model and RERI,API,S.And the interaction among multiple factors were analyzed by generalized multifactor dimensionality reduction(GMDR)method.Results:1.The association between HPV and CIN: The HPVs and HPV16 infection rate in CIN ?(42.4%,15.3%)and CIN ?/?(60%,41.7%)were higher than that in NC(21.3%,11.5%)group.HPVs infection lead to CIN ?,CIN ?/? are 2.576,4.732 times than HPV negative,and HPV16 infection lead to CIN ?/? is 4.086 times than HPV negative.With the aggravation of cervical lesions,the infection rate of HPVs and HPV16 both increased gradually(c2trend=18.607,P<0.001;c2trend=15.525,P<0.001).2.The association between HPV16 L1,L2 and CIN: the distribution of expression level about HPV16 L1 and L2 was not in common among each groups(H=8.689,P=0.013;H=6.878,P=0.032).The expression level of HPV16 L1 and L2 in CIN ?/? group was lower than CIN ?group,but the difference of HPV16 L1 and L2 protein between NC group and CIN ?,and NC group with CIN ?/? have no statistical significance.3.The association between hnRNP E2 and CIN: the distribution of expression level of hnRNP E2 was not in common among each groups(H=14.030,P=0.001).The expressionlevel of hnRNP E2 in CIN ?/? was lower than NC group,and the difference between CIN ?and CIN ?/? with NC group has not statistical significance.Low expression of hnRNP E2 lead to CIN ?/? is 4.531 times than high expression of hnRNP E2.With the aggravation of cervical lesions,the expression level of hnRNP E2 gradually decreased(c2trend=8.590,P=0.003).4.The association between folic acid and CIN: the distribution of serum folic acid was not in common among each groups(H=18.307,P<0.001).Serum folic acid levels in CIN ?and CIN ?/? were lower than NC group,and the difference between CIN ?and CIN ?/? has not statistical significance.Folate deficiency lead to CIN ?,CIN ?/? are3.684,7.320 times than folate adequate.With the aggravation of cervical lesions,folic acid level gradually decreased(c2trend=16.770,P<0.001).5.The relationship among folate,hnRNP E2,HPV16 L1 and L2 in cervical intraepithelial neoplasia: there were positive correlation between folate and HPV16 L1(r=0.149,P=0.046).Similar relationship can also be seen in hnRNP E2 and HPV16 L2(r=0.382,P=0.014),HPV16 L1 and L2(r=0.407,P=0.008).6.Analysis of the interaction of folic acid,hnRNP E2 and HPV16 among dfferent cervical intraepithelial neoplasia: there is positive additive interaction effect in CIN ?/?group between serum folic acid and HPV16,hnRNP E2 and HPV16.Use generalized multifactor dimensionality reduction methodinteraction(GMDR)to further analysis the interaction of folic acid,hnRNP E2,and HPV16 L1 and L2 in cervical intraepithelial neoplasia,the best interaction models of CIN?and CIN ?/? were model of folic acid,hnRNP E2,and HPV16 L1 and L2.Conclusion:1.Both HPVs and HPV16 infection can increase the risk of cervical intraepithelial neoplasia.So,it is helpful to prevent the occurrence of cervical cancer by developing HPV screening and preventing and controlling HPV infection.2.Folate deficiency appeared in the early days of cervical intraepithelial neoplasia.And there is synergistic effect between folate deficiency and HPV16 infection in CIN?/?.It indicates that early detection for folic acid level can provide important reference for prediction and assessment of lesions progress.3.The decreasion of expression levels about hnRNP E2,HPV16 L1 and L2 in CIN?/? group shows that low expression of hnRNP E2,HPV16 L1 and L2 can be used as a marker of high-grade cervical intraepithelial neoplasia.There is synergistic effect between the low expression of hnRNP E2 and HPV16 infection in CIN ?/? group.4.The interaction among folic acid deficiency,low expression of hnRNP E2 and HPV16 L1 and L2 may promote the risk of CIN ?and CIN ?/?.It shows that cervical intraepithelial neoplasia may be influenced by diet,genes and virus.