發(fā)布時(shí)間：2018-06-28 01:26

  本文選題：宮頸癌 + 甲基化��； 參考：《新疆醫(yī)科大學(xué)》2014年博士論文

【摘要】：目的：了解維吾爾族婦女型別特異性HPV持續(xù)感染的情況及危險(xiǎn)因素；探討維吾爾族婦女HPV持續(xù)感染、宮頸癌發(fā)生發(fā)展與HPV16L1、LCR甲基化的相關(guān)性。方法：收集2012年9月～2013年9月新疆自治區(qū)人民醫(yī)院婦科門診自愿接受宮頸癌機(jī)會(huì)性篩查維吾爾族婦女，利用HC-ⅡHPV DNA檢查和凱普HPV導(dǎo)流雜交分型技術(shù)確定HPV感染和型別，選擇篩查結(jié)果為HPV感染慢性宮頸炎的婦女和隨機(jī)選擇HPV陰性宮頸細(xì)胞學(xué)正常婦女各300例納入研究隊(duì)列，進(jìn)行巢式病例對(duì)照研究，通過(guò)問(wèn)卷調(diào)查和12個(gè)月隨訪，使用Logistic回歸分析其持續(xù)感染影響因素；選擇HPV16陽(yáng)性的篩查對(duì)象，分為一過(guò)感染組、持續(xù)感染組、CIN1組、CIN2-3組、宮頸癌組，使用焦磷酸測(cè)序技術(shù)，對(duì)HPV16L1、LCR區(qū)CpG位點(diǎn)的甲基化程度進(jìn)行定位和定量檢測(cè)，比較不同病理級(jí)別間HPV16甲基化的差異，使用ROC曲線分析評(píng)價(jià)HPV16各CpG位點(diǎn)甲基化對(duì)預(yù)測(cè)HPV持續(xù)感染、診斷CIN2+病變的價(jià)值；利用免疫組織化學(xué)技術(shù)檢測(cè)HPV16殼蛋白L1的表達(dá)，利用多重實(shí)時(shí)定量PCR技術(shù)檢測(cè)分析HPV16與宿主基因組的整合狀態(tài)，分析L1表達(dá)、HPV16整合游離狀態(tài)與HPV16甲基化的關(guān)系。結(jié)果：(1)慢性宮頸炎的維吾爾族婦女型別特異性HPV持續(xù)感染率為25.5%（65/255），持續(xù)感染率前5位型別依次為HPV16(48.00%)、HPV18(31.03%)、HPV58(28.30%)、HPV52(23.40%)、HPV45(21.43%)；HPV16陽(yáng)性相對(duì)非HPV16持續(xù)感染危險(xiǎn)增加(OR:4.81,95%CI:2.63-8.18)；極高病毒載量相對(duì)于低病毒載量持續(xù)感染的風(fēng)險(xiǎn)增加(OR:2.40,95%CI:1.11-5.15)；相對(duì)一過(guò)感染組，HPV持續(xù)感染的危險(xiǎn)因素包括絕經(jīng)(OR:1.83,95%CI:1.01-2.62)和不使用避孕套(OR:2.18,95%CI:1.71-3.46)。(2)HPV16L113個(gè)CpG位點(diǎn)甲基化率隨著病情進(jìn)展，甲基化率呈上升趨勢(shì)，進(jìn)行多組比較，各組13個(gè)位點(diǎn)的甲基化率比較有統(tǒng)計(jì)學(xué)差異P0.001，并且13個(gè)CpG位點(diǎn)高甲基化均增加患CIN2+病變的風(fēng)險(xiǎn)， OR值最大CpG位點(diǎn)為6650(OR:9.89,95%CI:3.57-27.44)；HPV16L113個(gè)CpG位點(diǎn)診斷CIN2+的AUCs在0.756-0.862之間，最高診斷價(jià)值的是6650位點(diǎn)，AUC為0.862；持續(xù)感染組與一過(guò)感染組比較，位點(diǎn)6389、6457、6581、6650、6796、7034高甲基化增加HPV16持續(xù)感染的風(fēng)險(xiǎn)，P0.05，OR值最高的CpG位點(diǎn)為6389(OR:13.33,95%CI:3.95-28.08)，，預(yù)測(cè)HPV持續(xù)感染AUCs在0.656-0.943之間，最高AUC是位點(diǎn)6389，AUC為0.943。HPV16LCR啟動(dòng)子區(qū)CpG位點(diǎn)31、37、43、52和58甲基化率在各個(gè)宮頸病變組間差異有統(tǒng)計(jì)學(xué)意義，并且隨著病情進(jìn)展，甲基化率呈現(xiàn)上升趨勢(shì)，P=0.001；這些位點(diǎn)的高甲基化患CIN2+病變的風(fēng)險(xiǎn)增加，P0.05；其中相關(guān)性最明顯的位點(diǎn)為58(OR:5.71,95%CI:2.54-12.84)，以上位點(diǎn)高甲基化率診斷CIN2+的AUCs在0.640-0.848之間，診斷價(jià)值最大的位點(diǎn)為58，AUCs為0.848；在CIN2-3組、宮頸癌組HPV16LCR啟動(dòng)子區(qū)、E2BS3、E2BS4甲基化率隨著結(jié)合狀態(tài)比例升高，甲基化率呈下降趨勢(shì)，差異有統(tǒng)計(jì)學(xué)意義，P0.05；宮頸癌組整合狀態(tài)下的甲基化水平仍在11.50%到14.25%之間。結(jié)論：HPV16陽(yáng)性、極高病毒載量、絕經(jīng)、不使用避孕套是維吾爾族婦女HPV持續(xù)感染的危險(xiǎn)因素，應(yīng)加強(qiáng)該人群的隨訪監(jiān)測(cè)；HPV16L1區(qū)甲基化對(duì)預(yù)測(cè)HPV持續(xù)感染、宮頸癌前病變進(jìn)展具有較高的價(jià)值；HPV16LCR啟動(dòng)子區(qū)，特別是E2BS3、E2BS4的甲基化水平與宮頸病變進(jìn)展具有相關(guān)性，提示啟動(dòng)子區(qū)甲基化在調(diào)節(jié)病毒基因轉(zhuǎn)錄中發(fā)揮重要作用。
[Abstract]:Objective: to understand the status and risk factors of HPV continuous infection in Uygur women, and to explore the correlation between the continuous infection of HPV in Uygur women, the development of cervical cancer and the HPV16L1 and LCR methylation. Methods: collect the opportunistic screening of cervical cancer from September 2012 to September 2013 in the gynecological clinic of the people's Hospital of Xinjiang Autonomous Region The Uygur women were examined by HC- II HPV DNA examination and KP HPV diversion cross typing technique to determine HPV infection and type. 300 women with HPV infected chronic cervicitis and randomly selected HPV negative cervical cytology were included in the cohort, and a nested case control study was conducted. A questionnaire survey and 12 cases were conducted. Logistic regression was used to analyze the influencing factors of persistent infection, and HPV16 positive screening subjects were selected to be divided into one over infection group, continuous infection group, CIN1 group, CIN2-3 group and cervical cancer group, using pyrosequencing technology to locate and quantify the methylation of CpG loci in HPV16L1 and LCR region, and compare the HPV1 between different pathological grades. The difference between 6 methylation and ROC curve analysis was used to evaluate the value of HPV16 CpG locus methylation in predicting HPV continuous infection and diagnosis of CIN2+ lesions. The expression of HPV16 shell protein L1 was detected by immunohistochemical technique, and the integrated state of HPV16 and host genome was detected by multiple real-time quantitative PCR technique, and L1 expression and HPV16 integration were analyzed. The relationship between free state and HPV16 methylation. Results: (1) the persistent infection rate of Uygur specific HPV in chronic cervicitis was 25.5% (65/255), and the first 5 types of persistent infection rate were HPV16 (48%), HPV18 (31.03%), HPV58 (28.30%), HPV52 (23.40%), HPV45 (21.43%); HPV16 positive relative to non HPV16 infection increased (OR:). 4.81,95%CI:2.63-8.18); the risk of high viral load relative to persistent infection of low viral load increased (OR:2.40,95%CI:1.11-5.15); relative to one over infection group, the risk factors for persistent infection of HPV include Menopause (OR:1.83,95%CI:1.01-2.62) and non use of condoms (OR: 2.18,95%CI:1.71-3.46). (2) the methylation rate of HPV16L113 CpG sites along with the rate of methylation The methylation rate was on the rise. The methylation rate of the 13 loci in each group had a statistically significant difference P0.001, and the 13 CpG loci hypermethylation increased the risk of CIN2+ disease, and the maximum CpG site of the OR value was 6650 (OR:9.89,95%CI:3.57-27.44), and AUCs of the HPV16L113 CpG site for the diagnosis of CIN2+ was 0.756-0.862. The highest diagnostic value was 6650 loci, AUC was 0.862, and in the continuous infection group, the 638964576581665067967034 hypermethylation at the site increased the risk of HPV16 persistent infection, P0.05, the highest OR value of the CpG site was 6389 (OR:13.33,95%CI:3.95-28.08), and the pretest HPV continued to infect AUCs in 0.656-0.943, the highest AUC was Loci 6389, AUC 0.943.HPV16LCR promoter region CpG locus 31,37,43,52 and 58 methylation rates were statistically significant differences between the various cervical lesions, and as the condition progressed, the rate of methylation increased, P=0.001; the hypermethylation of these sites increased the risk of CIN2+ lesions, P0.05; the most significant correlation was in the loci of these sites. 58 (OR:5.71,95%CI:2.54-12.84), the higher methylation rate of the above loci diagnosis of CIN2+ AUCs between 0.640-0.848, the most diagnostic value of the site is 58, AUCs is 0.848. In the CIN2-3 group, the HPV16LCR promoter region of the cervical cancer group, E2BS3, E2BS4 methylation rate increases with the proportion of binding state, the methylation rate decreases, the difference is statistically significant, P0., the difference is statistically significant, P0. difference is statistically significant, P0. difference is statistically significant, P0. difference is statistically significant, P0. difference is statistically significant 05; the methylation level in the cervical cancer group is still between 11.50% and 14.25%. Conclusion: HPV16 positive, high viral load, menopause, and no condom use is a risk factor for the continuous infection of Uygur women, and the follow-up monitoring should be strengthened in this group. HPV16L1 region methylation is used to predict HPV continuous infection and advance of cervical precancerous lesions. It is of high value; the HPV16LCR promoter region, especially the E2BS3, the level of methylation of E2BS4 is associated with the progression of cervical lesions, suggesting that promoter methylation plays an important role in regulating viral gene transcription.
【學(xué)位授予單位】：新疆醫(yī)科大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2014
【分類號(hào)】：R737.33

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