hnRNP E1与HPV16早期基因E2和E6在宫颈病变中的作用及交互效应

发布时间：2018-04-17 23:17

本文选题：hnRNP + E1　；参考：《山西医科大学》2017年硕士论文

【摘要】：目的:人乳头瘤病毒(human papilloma virus,HPV),特别是高危型HPV(HR-HPV)16持续感染是宫颈癌及其癌前病变最主要的病因因素,HPV DNA与宿主细胞DNA整合是病毒致癌作用的重要机制。HPV16早期基因E2和E6是与病毒致癌密切相关的基因,E2是HPV整合的关键基因,E6是病毒癌基因。核不均一核糖核蛋白E1(heterogeneous nuclear ribonucleoprotein,hnRNP E1)在基因转录和转录后调控中具有独特作用,特别值得关注的是,hnRNP E1特有的KH域提供了与HPV16基因发生特异性结合的独特分子结构,但hnRNP E1在宫颈病变中的作用及与HPV16E2和E6的关系尚不清楚。本次研究以不同宫颈病变患者为研究对象,明确其HPV感染状态,检测不同级别宫颈组织中hnRNP E1蛋白表达水平,以及与HPV16整合相关基因E2蛋白表达水平和致癌基因E6蛋白表达水平。以期揭示hnRNP E1与HPV16早期基因E2和E6在宫颈癌变进展中的作用及相互关系,为进行HPV致宫颈癌作用机制的研究提供新思路。方法:从课题组于2014年6月至9月在山西省介休市建立的自然人群宫颈病变队列中,分别选取正常宫颈妇女(NC)56例,低度宫颈上皮内瘤变(CINⅠ)患者58例和高度宫颈上皮内瘤变(CINⅡ/Ⅲ)患者50例,以及2015年11月至2016年5月在山西省肿瘤医院就诊的宫颈鳞状细胞癌(SCC)病人40例作为研究对象,所有的研究对象都是经液基薄层细胞检测技术(TCT)筛查、阴道镜检查、最终由病理学确诊的。在获得知情同意的情况下,采用结构式问卷收集全部研究对象的人口学特征、生活习惯、个人卫生习惯和生殖情况等资料,同时采集全部研究对象宫颈组织活检标本和宫颈脱落细胞。采用导流杂交法检测HPV感染状况,Western blot检测hnRNP E1以及HPV16早期基因E2和E6蛋白表达水平。应用SPSS16.0软件进行相关资料的Kruskal-Wallis H检验、Bonferroni检验、χ2检验、χ2趋势检验、Spearman秩相关分析、logistic回归和因子分析,应用相加模型、相乘模型及广义多因子降维(GMDR)模型进行交互作用评价。结果:1.HPVs/HPV16感染与宫颈病变的关系:HPVs感染率(41.38%、58.00%、87.5%)和HPV16感染率(15.52%、40.00%、67.50%)在CINⅠ、CINⅡ/Ⅲ和SCC组均高于NC组的感染率(16.07%、8.93%),并随着宫颈病变严重程度加重均呈现升高趋势。HPVs感染(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))和HPV16感染(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))与宫颈病变之间均呈正关联。2.hnRNP E1与宫颈病变的关系:hnRNP E1表达量在不同宫颈病变组间差异有统计学意义(H=9.979,P=0.019),随着宫颈病变严重程度加重呈现降低趋势(χ2趋势=9.50,P=0.002),hnRNP E1表达量与CINⅡ/Ⅲ和SCC之间均呈负关联。hnRNP E1低表达与HPV16感染在宫颈病变中存在正相加交互作用,而相乘交互作用未显示有统计学意义。3.HPV16早期基因E2和E6与宫颈病变的关系:HPV16 E2表达水平(H=16.20,P=0.001)与HPV16 E6表达水平(H=15.44,P=0.001)在不同宫颈病变组间差异均有统计学意义。HPV16 E2蛋白表达量在SCC组和CINⅡ/Ⅲ组均低于NC组,SCC组低于CINⅠ组,差异均有统计学意义,其它组间差异均无统计学意义。HPV16 E6蛋白表达量仅在SCC组与CINⅠ组差异有统计学意义,其余任意两组之间差异均无统计学意义。E2/E6比值与宫颈病变程度呈现负相关。4.hnRNP E1与HPV16早期基因E2和E6在宫颈病变中的交互作用:应用广义多因子降维模型(GMDR)分析显示,hnRNP E1低表达/HPV16 E2低表达/HPV16E6高表达在CINⅡ/Ⅲ和SCC组存在交互作用,而在CINⅠ组未发现类似交互作用存在(P0.05)。结论:1.HPVs/HPV16感染是宫颈病变发生的危险因素,预防和控制HPVs/HPV16感染可降低宫颈病变发生的风险。2.hnRNP E1低表达可增加CINⅡ/Ⅲ和SCC的发生风险,提示hnRNP E1表达可以反映宫颈病变进展情况,hnRNP E1低表达可能是宫颈病变加重的有效指标,可以作为高度宫颈病变和癌变的诊断和预测依据,且hnRNP E1低表达与HPV16感染在宫颈病变的发生中可能存在协同作用。3.HPV16整合关键基因E2蛋白低表达与主要致癌基因E6蛋白高表达是SCC发生的危险因素,是宫颈癌变的有效指标,可以作为SCC的诊断依据。E2/E6比值与宫颈病变程度呈负相关,可以反映宫颈病变的进展情况。4.hnRNP E1低表达/HPV16 E2低表达/HPV16 E6高表达在宫颈病变中存在交互作用,三者共同存在时的作用大于单独存在时作用的和,提示宫颈病变的发生可能存在病毒与基因之间的协同作用。hnRNP E1可以作为防治宫颈病变的干预靶点,预防和控制HPV16感染,降低两者形成交互效应的机会,可使宫颈病变发生的风险降低。
[Abstract]:Objective: human papillomavirus (human papilloma, virus, HPV), especially the high-risk HPV (HR-HPV) 16 is the persistent infection of cervical cancer and precancerous lesions is the main etiological factors of HPV, DNA and DNA virus host cell integration is the important mechanism of.HPV16 carcinogenesis early gene E2 and E6 are closely related with viral carcinogenesis the E2 gene is the key gene of HPV integration, E6 is a viral oncogene. Heterogeneous nuclear ribonucleoprotein E1 (heterogeneous nuclear ribonucleoprotein, hnRNP E1) has a unique role in gene transcription and post transcription regulation, of particular concern is that the KH domain of hnRNP E1 provides a unique unique molecular structure specificity combined with the HPV16 gene, but the relationship between hnRNP and E1 in cervical lesions with HPV16E2 and E6 is not clear. In this study, patients with different cervical lesions as the research object, the HPV infection, detection The expression of hnRNP E1 protein level in the same level of cervical tissues, and the integration of HPV16 and related gene expression levels of E2 protein and E6 oncogene protein expression level of E6 hnRNP and E2 revealed early gene E1 and HPV16 in cervical cancer progression and the relationship in order to provide new ideas for the study of mechanism of HPV induced cervical cancer methods: the cervical lesion group from natural population cohort from June 2014 to September in Shanxi city of Jiexiu province established in selected women of normal cervix (NC) in 56 cases of low-grade cervical intraepithelial neoplasia (CIN 1) and 58 patients with high-grade cervical intraepithelial neoplasia (CIN II / III) 50 cases in November 2015 to May 2016, as well as in cervical squamous cell carcinoma from the tumor hospital of Shanxi province (SCC) in 40 patients as the research object, all of the subjects by ThinPrep Pap test (TCT) screening, colposcopy, The final pathological diagnosis. Informed consent was obtained under the condition of demographic characteristics, using a structured questionnaire to collect all the objects of study habits, personal hygiene and reproductive data collected at the same time, all the research object of cervical tissue biopsy specimens and cervical exfoliated cells. Infection was detected by HPV hybridization method, Western blot detection of hnRNP E1 and HPV16 early gene E2 and E6 protein expression level. Kruskal-Wallis H test, SPSS16.0 software was used to analyze the related data of Bonferroni test, x 2 x 2 test, trend test, Spearman rank correlation analysis, logistic regression analysis and factor, application of additive model, multiplication model and generalized multifactor dimensionality reduction (GMDR) of the interaction of the evaluation model. Results: the relationship between 1.HPVs/HPV16 infection and cervical lesions: the infection rate of HPVs (41.38%, 58%, 87.5%) and the infection rate of HPV16 (15.52%, 40%, 67.5 0%) in CIN I and CIN II / III and SCC groups were higher than NC group. The infection rate (16.07%, 8.93%), and with the severity of cervical lesions increased.HPVs infection (CIN 1: a OR=3.62 (1.41-9.29); CIN II / III: a OR=6.44 (2.48-16.72); SCC:a OR=28.32 (5.24-98.46)) and HPV16 infection (CIN 1: a OR=1.73 (0.51-5.86); CIN II / III: a OR=5.96 (1.92-18.49); SCC:a OR=13.96 (3.33-58.50)) and the relationship between cervical lesions were positively associated with.2.hnRNP E1 and hnRNP E1 expression in cervical lesions: there was significant difference in different cervical lesion group (H=9.979. P=0.019), with the severity of cervical lesions increased decreased (x 2 =9.50 trend, P=0.002), hnRNP expression of E1 and CIN II / III and SCC were negatively associated with.HnRNP low expression of E1 and HPV16 infection in cervical lesions in positive additive interaction, and multiplicative interaction did not show statistical The relationship between significance.3.HPV16 early gene E2 and E6 cervical lesions and HPV16: the expression level of E2 (H=16.20, P=0.001) level and HPV16 expression of E6 (H=15.44, P=0.001) were statistically significant differences in.HPV16 E2 protein expression in different cervical lesion group were lower than those of NC group in SCC group and CIN II / III group, SCC group less than CIN group, there were statistically significant differences, the other groups had no statistically significant difference between the.HPV16 expression of E6 only showed a significant difference in SCC group and CIN group, the rest of any differences between the two groups were not statistically significant interaction between.E2/E6 ratio and the degree of cervical lesions showed a negative correlation with.4.hnRNP E1 HPV16 early gene E2 and E6 in cervical lesions: application of generalized multifactor dimensionality reduction model (GMDR) analysis showed that hnRNP low expression of E1 low expression of /HPV16 high expression of E2 /HPV16E6 interaction in CIN II / III and group SCC, and CIN in group I There is no similar interaction (P0.05). Conclusion: 1.HPVs/HPV16 infection is a risk factor for the development of cervical lesions, the prevention and control of HPVs/HPV16 infection can reduce the risk of cervical lesions.2.hnRNP low expression of E1 can increase the risk of SCC and CIN II /, suggesting that hnRNP E1 expression can reflect the progression of cervical lesions, the low expression of hnRNP E1 may be a useful indicator of cervical lesions increased, can be used as a basis for prediction and diagnosis of high cervical lesions and cancer, and hnRNP low expression of E1 and HPV16 infection in cervical lesions may exist in synergy with.3.HPV16 integration key gene low expression of E2 protein and gene expression of E6 protein is the risk factors of SCC that is a valid indicator of cervical cancer, SCC can be used as a basis for the diagnosis of.E2/E6 ratio was negatively correlated with the degree of cervical lesions, can reflect the development situation of cervical lesions .4.hnRNP low expression of E1 low expression of /HPV16 E2 high expression of /HPV16 E6 interaction in cervical lesions, the three co existing effect than when they are alone and the role of occurrence of cervical lesions, suggesting possible synergistic effects of.HnRNP E1 between the virus and the gene can be used as the intervention target for the prevention and treatment of cervical disease, prevention and control of HPV16 infection, reduce both the formation of interaction opportunities, can make the risk of cervical lesions decreased.

【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R737.33

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