本文選題：hnRNP + E1��； 參考：《山西醫(yī)科大學(xué)》2017年碩士論文

【摘要】：目的:人乳頭瘤病毒(human papilloma virus,HPV),特別是高危型HPV(HR-HPV)16持續(xù)感染是宮頸癌及其癌前病變最主要的病因因素,HPV DNA與宿主細(xì)胞DNA整合是病毒致癌作用的重要機制。HPV16早期基因E2和E6是與病毒致癌密切相關(guān)的基因,E2是HPV整合的關(guān)鍵基因,E6是病毒癌基因。核不均一核糖核蛋白E1(heterogeneous nuclear ribonucleoprotein,hnRNP E1)在基因轉(zhuǎn)錄和轉(zhuǎn)錄后調(diào)控中具有獨特作用,特別值得關(guān)注的是,hnRNP E1特有的KH域提供了與HPV16基因發(fā)生特異性結(jié)合的獨特分子結(jié)構(gòu),但hnRNP E1在宮頸病變中的作用及與HPV16E2和E6的關(guān)系尚不清楚。本次研究以不同宮頸病變患者為研究對象,明確其HPV感染狀態(tài),檢測不同級別宮頸組織中hnRNP E1蛋白表達(dá)水平,以及與HPV16整合相關(guān)基因E2蛋白表達(dá)水平和致癌基因E6蛋白表達(dá)水平。以期揭示hnRNP E1與HPV16早期基因E2和E6在宮頸癌變進展中的作用及相互關(guān)系,為進行HPV致宮頸癌作用機制的研究提供新思路。方法:從課題組于2014年6月至9月在山西省介休市建立的自然人群宮頸病變隊列中,分別選取正常宮頸婦女(NC)56例,低度宮頸上皮內(nèi)瘤變(CINⅠ)患者58例和高度宮頸上皮內(nèi)瘤變(CINⅡ/Ⅲ)患者50例,以及2015年11月至2016年5月在山西省腫瘤醫(yī)院就診的宮頸鱗狀細(xì)胞癌(SCC)病人40例作為研究對象,所有的研究對象都是經(jīng)液基薄層細(xì)胞檢測技術(shù)(TCT)篩查、陰道鏡檢查、最終由病理學(xué)確診的。在獲得知情同意的情況下,采用結(jié)構(gòu)式問卷收集全部研究對象的人口學(xué)特征、生活習(xí)慣、個人衛(wèi)生習(xí)慣和生殖情況等資料,同時采集全部研究對象宮頸組織活檢標(biāo)本和宮頸脫落細(xì)胞。采用導(dǎo)流雜交法檢測HPV感染狀況,Western blot檢測hnRNP E1以及HPV16早期基因E2和E6蛋白表達(dá)水平。應(yīng)用SPSS16.0軟件進行相關(guān)資料的Kruskal-Wallis H檢驗、Bonferroni檢驗、χ2檢驗、χ2趨勢檢驗、Spearman秩相關(guān)分析、logistic回歸和因子分析,應(yīng)用相加模型、相乘模型及廣義多因子降維(GMDR)模型進行交互作用評價。結(jié)果:1.HPVs/HPV16感染與宮頸病變的關(guān)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組均高于NC組的感染率(16.07%、8.93%),并隨著宮頸病變嚴(yán)重程度加重均呈現(xiàn)升高趨勢。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))與宮頸病變之間均呈正關(guān)聯(lián)。2.hnRNP E1與宮頸病變的關(guān)系:hnRNP E1表達(dá)量在不同宮頸病變組間差異有統(tǒng)計學(xué)意義(H=9.979,P=0.019),隨著宮頸病變嚴(yán)重程度加重呈現(xiàn)降低趨勢(χ2趨勢=9.50,P=0.002),hnRNP E1表達(dá)量與CINⅡ/Ⅲ和SCC之間均呈負(fù)關(guān)聯(lián)。hnRNP E1低表達(dá)與HPV16感染在宮頸病變中存在正相加交互作用,而相乘交互作用未顯示有統(tǒng)計學(xué)意義。3.HPV16早期基因E2和E6與宮頸病變的關(guān)系:HPV16 E2表達(dá)水平(H=16.20,P=0.001)與HPV16 E6表達(dá)水平(H=15.44,P=0.001)在不同宮頸病變組間差異均有統(tǒng)計學(xué)意義。HPV16 E2蛋白表達(dá)量在SCC組和CINⅡ/Ⅲ組均低于NC組,SCC組低于CINⅠ組,差異均有統(tǒng)計學(xué)意義,其它組間差異均無統(tǒng)計學(xué)意義。HPV16 E6蛋白表達(dá)量僅在SCC組與CINⅠ組差異有統(tǒng)計學(xué)意義,其余任意兩組之間差異均無統(tǒng)計學(xué)意義。E2/E6比值與宮頸病變程度呈現(xiàn)負(fù)相關(guān)。4.hnRNP E1與HPV16早期基因E2和E6在宮頸病變中的交互作用:應(yīng)用廣義多因子降維模型(GMDR)分析顯示,hnRNP E1低表達(dá)/HPV16 E2低表達(dá)/HPV16E6高表達(dá)在CINⅡ/Ⅲ和SCC組存在交互作用,而在CINⅠ組未發(fā)現(xiàn)類似交互作用存在(P0.05)。結(jié)論:1.HPVs/HPV16感染是宮頸病變發(fā)生的危險因素,預(yù)防和控制HPVs/HPV16感染可降低宮頸病變發(fā)生的風(fēng)險。2.hnRNP E1低表達(dá)可增加CINⅡ/Ⅲ和SCC的發(fā)生風(fēng)險,提示hnRNP E1表達(dá)可以反映宮頸病變進展情況,hnRNP E1低表達(dá)可能是宮頸病變加重的有效指標(biāo),可以作為高度宮頸病變和癌變的診斷和預(yù)測依據(jù),且hnRNP E1低表達(dá)與HPV16感染在宮頸病變的發(fā)生中可能存在協(xié)同作用。3.HPV16整合關(guān)鍵基因E2蛋白低表達(dá)與主要致癌基因E6蛋白高表達(dá)是SCC發(fā)生的危險因素,是宮頸癌變的有效指標(biāo),可以作為SCC的診斷依據(jù)。E2/E6比值與宮頸病變程度呈負(fù)相關(guān),可以反映宮頸病變的進展情況。4.hnRNP E1低表達(dá)/HPV16 E2低表達(dá)/HPV16 E6高表達(dá)在宮頸病變中存在交互作用,三者共同存在時的作用大于單獨存在時作用的和,提示宮頸病變的發(fā)生可能存在病毒與基因之間的協(xié)同作用。hnRNP E1可以作為防治宮頸病變的干預(yù)靶點,預(yù)防和控制HPV16感染,降低兩者形成交互效應(yīng)的機會,可使宮頸病變發(fā)生的風(fēng)險降低。
[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.

【學(xué)位授予單位】：山西醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R737.33

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