维吾尔族妇女HPV持续感染、宫颈癌发生发展与HPV16 L1、LCR甲基化的相关性研究

发布时间：2018-06-28 01:26

本文选题：宫颈癌 + 甲基化　；参考：《新疆医科大学》2014年博士论文

【摘要】：目的：了解维吾尔族妇女型别特异性HPV持续感染的情况及危险因素；探讨维吾尔族妇女HPV持续感染、宫颈癌发生发展与HPV16L1、LCR甲基化的相关性。方法：收集2012年9月～2013年9月新疆自治区人民医院妇科门诊自愿接受宫颈癌机会性筛查维吾尔族妇女，利用HC-ⅡHPV DNA检查和凯普HPV导流杂交分型技术确定HPV感染和型别，选择筛查结果为HPV感染慢性宫颈炎的妇女和随机选择HPV阴性宫颈细胞学正常妇女各300例纳入研究队列，进行巢式病例对照研究，通过问卷调查和12个月随访，使用Logistic回归分析其持续感染影响因素；选择HPV16阳性的筛查对象，分为一过感染组、持续感染组、CIN1组、CIN2-3组、宫颈癌组，使用焦磷酸测序技术，对HPV16L1、LCR区CpG位点的甲基化程度进行定位和定量检测，比较不同病理级别间HPV16甲基化的差异，使用ROC曲线分析评价HPV16各CpG位点甲基化对预测HPV持续感染、诊断CIN2+病变的价值；利用免疫组织化学技术检测HPV16壳蛋白L1的表达，利用多重实时定量PCR技术检测分析HPV16与宿主基因组的整合状态，分析L1表达、HPV16整合游离状态与HPV16甲基化的关系。结果：(1)慢性宫颈炎的维吾尔族妇女型别特异性HPV持续感染率为25.5%（65/255），持续感染率前5位型别依次为HPV16(48.00%)、HPV18(31.03%)、HPV58(28.30%)、HPV52(23.40%)、HPV45(21.43%)；HPV16阳性相对非HPV16持续感染危险增加(OR:4.81,95%CI:2.63-8.18)；极高病毒载量相对于低病毒载量持续感染的风险增加(OR:2.40,95%CI:1.11-5.15)；相对一过感染组，HPV持续感染的危险因素包括绝经(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个CpG位点甲基化率随着病情进展，甲基化率呈上升趋势，进行多组比较，各组13个位点的甲基化率比较有统计学差异P0.001，并且13个CpG位点高甲基化均增加患CIN2+病变的风险， OR值最大CpG位点为6650(OR:9.89,95%CI:3.57-27.44)；HPV16L113个CpG位点诊断CIN2+的AUCs在0.756-0.862之间，最高诊断价值的是6650位点，AUC为0.862；持续感染组与一过感染组比较，位点6389、6457、6581、6650、6796、7034高甲基化增加HPV16持续感染的风险，P0.05，OR值最高的CpG位点为6389(OR:13.33,95%CI:3.95-28.08)，，预测HPV持续感染AUCs在0.656-0.943之间，最高AUC是位点6389，AUC为0.943。HPV16LCR启动子区CpG位点31、37、43、52和58甲基化率在各个宫颈病变组间差异有统计学意义，并且随着病情进展，甲基化率呈现上升趋势，P=0.001；这些位点的高甲基化患CIN2+病变的风险增加，P0.05；其中相关性最明显的位点为58(OR:5.71,95%CI:2.54-12.84)，以上位点高甲基化率诊断CIN2+的AUCs在0.640-0.848之间，诊断价值最大的位点为58，AUCs为0.848；在CIN2-3组、宫颈癌组HPV16LCR启动子区、E2BS3、E2BS4甲基化率随着结合状态比例升高，甲基化率呈下降趋势，差异有统计学意义，P0.05；宫颈癌组整合状态下的甲基化水平仍在11.50%到14.25%之间。结论：HPV16阳性、极高病毒载量、绝经、不使用避孕套是维吾尔族妇女HPV持续感染的危险因素，应加强该人群的随访监测；HPV16L1区甲基化对预测HPV持续感染、宫颈癌前病变进展具有较高的价值；HPV16LCR启动子区，特别是E2BS3、E2BS4的甲基化水平与宫颈病变进展具有相关性，提示启动子区甲基化在调节病毒基因转录中发挥重要作用。
[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.
【学位授予单位】：新疆医科大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R737.33

【相似文献】