Effects Of HnRNP E1 And HPV16 Early Gene E2 And E6 In Cervical Lesions And Their Interaction

Objective: Human papillomavirus(HPV),particularly high-risk HPV(HR-HPV)16 persistent infection is a key etiology of cervical cancer and its precancerous lesions,HPV DNA and host cell DNA integration is important mechanism of viral carcinogenesis.HPV16 early genes E2 and E6 are closely related to its carcinogenicity,E2 is HPV integration key gene,E6 is virus oncogene.Heterogeneous nuclear ribonucleoprotein E1(hn RNP E1)plays a unique role in the regulation of gene transcription and post transcription,It is noteworthy that the KH domain of hn RNP E1 provides unique molecular structure that binds specifically to HPV16 gene,but the role of hn RNP E1 in cervical lesions and the relationship with HPV16 E2 and E6 is still unknown.In this study,different cervical lesions patients were selected as the research object,HPV infection status were detected,hn RNP E1 protein expression levels,HPV16 integrated key gene E2 and oncogene E6 expression levels were measured,this study reveals the role and relationship of hn RNP E1 and HPV16 early gene E2 and E6 in the development of cervical cancer,this provides a novel insight into mechanisms of HPV causing cervical carcinogenesis.Methods: The study object were 56 women with normal cervix(NC),58 patients with low-grade cervical intraepithelial neoplasm(CINⅠ)and 50 patients with high-grade cervical intraepithelial neoplasm(CINⅡ/Ⅲ)collected from the population cervical lesions cohort that established by our research group in Jiexiu of Shanxi province form June to September of 2014,and 40 patients with cervical squamous cell carcinoma(SCC)collected from Shanxi Tumor Hospital from November of 2015 to May of 2016,all the participants were experienced Thinprep Cytologic Test(TCT),colposcopy uterus screening and ultimately diagnosed by pathology.After obtaining written informed consent,the information of demographic characteristics,lifestyle,personal hygiene behavior and reproductive status were collected using structured questionnaire,simultaneously cervical tissues and cervical cells were obtained from all the participants.HPV infection status were detected by flow-through hybridization,hn RNP E1 and HPV16 early gene E2 and E6 protein expression levels were measured by western blot.The data were analyzed with the method of Kruskal-Wallis H test,Bonferroni test,chi-square test and trend of chi-square test,spearman rank correlation,multinomial logistic and factorial analysis by SPSS(version 16.0)statistical software.Additive model,multiplicative model and generalized multifactor dimensionality reduction(GMDR)were used for interaction effect analysis.Results: 1.Relationship between HPVs/HPV16 infection and cervical lesions:The infection rate of HPVs(41.38%,58.00%,87.5%)and HPV16(15.52%,40.00% and 67.50%)in the CINⅠ,CINⅡ/Ⅲ and SCC groups were all higher than the infection rate in NC group(16.07%,8.93%),and showed an increasing trend with the severity of cervical lesions.HPVs infection(CINⅠ: a OR=3.62(1.41-9.29);CINⅡ/Ⅲ: a OR=6.44(2.48-16.72);SCC: a OR=28.32(5.24-98.46))and HPV16 infection(CINⅠ: a OR=1.73(0.51-5.86);CINⅡ/Ⅲ: a OR=5.96(1.92-18.49);SCC: a OR=13.96(3.33-58.50))were positively associated with cervical lesions.2.Relationship between hn RNP E1 protein expression levels and cervical lesions: The average expression levels of hn RNP E1 protein were significantly different in cervical lesions groups(H=9.979,P=0.019),and showed decreasing trend with the severity of cervical lesions(χ2趋势=9.50,P=0.002),hn RNP E1 expression levels are negatively associated with CINⅡ/Ⅲ and SCC.hn RNP E1 low expression and HPV16 infection had additive interaction in cervical lesions,but multiplicative interaction had not statistical difference.3.Relationship between HPV16 early gene E2 and E6 and cervical lesions: The expression levels of HPV16 E2(H=16.20,P=0.001)and HPV16 E6(H=15.44,P=0.001)were significant different in different cervical lesions groups.Expression levels of HPV16 E2 protein in the SCC and CINⅡ/Ⅲ groups were lower than in NC group,and in SCC group was lower than in CINI group,the differences were statistically significant,but there was no significant difference between the other groups.The expression levels of HPV16 E6 protein in SCC group and CINI group were statistically significant,and there was no significant difference between the other groups.E2 / E6 ratio and the degree of cervical lesions was negatively correlated.4.The interaction of hn RNP E1 and HPV16 early gene E2 and E6 in cervical lesions: The generalized multifactor dimensionality reduction(GMDR)showed that the best interaction model in groups of CINⅡ/Ⅲ and SCC was hn RNP E1 low expression/HPV16 E2 low expression/HPV16 E6 high expression,but there had not similar interaction in the group of CINⅠ(P>0.05).Conclusion: 1.HPVs/HPV16 infection is the risk factor of cervical lesions,prevention and control of HPV s/HPV16 infection reduce the risk of cervical lesions.2.hn RNP E1 low expression may increase the risk of CINⅡ/Ⅲ and SCC,suggesting that hn RNP E1 expression can reflect the progression of cervical lesions,hn RNP E1 low expression may be useful indicator of cervical lesions weighted and may regard as diagnosis and prediction basis of high-grade cervical lesions and cancer,and there may have synergistic action between hn RNP E1 protein low expression and HPV16 infection.3.Low expression of HPV16 integrated key gene E2 and high expression of HPV16 major oncogene E6 is risk factor of SCC,and may be useful indicator of cervical cancer and may regarded as diagnosis basis of SCC.E2/E6 ratio is negatively correlated with the degree of cervical lesions,and can reflect the progress of cervical lesions.4.There is interaction between hn RNP E1 low expression,HPV16 E2 low expression and HPV16 E6 high expression in cervical lesions,suggesting that the happening of cervical lesions may exist the synergistic action of viruses and genes.hn RNP E1 may be used as a target for the prevention and treatment of cervical lesions,prevention and control of HPV16 infection may reduce the chance to form interaction and reduce the risk of cervical lesions.