宫颈癌筛查新方法及标志物的临床应用价值初探

发布时间：2018-06-15 15:42

本文选题：宫颈病变 + 叶酸受体介导上皮特殊染色液　；参考：《中国人民解放军医学院》2016年博士论文

【摘要】：目的：评价叶酸受体介导上皮特殊染色液、肿瘤细胞周期和表达相关蛋白和HPV E6/E7 mRNA分型检测在宫颈癌筛查中的应用价值,明确叶酸受体、肿瘤细胞周期和表达相关蛋白在宫颈病变组织和宫颈脱落细胞中的表达。方法：1、对231例门诊欲行宫颈癌筛查者,前瞻性行液基细胞学检查(TCT)、人乳头瘤病毒检测(HPV)和叶酸受体介导上皮特殊染色液(FRD)检查,并以病理诊断为金标准,计算并对比TCT.HPV和FRD的敏感度、特异度等诊断价值指标。2、通过荧光定量PCR方法,对153例宫颈脱落细胞行HPV DNA分型、HPV E6/E7 mRNA分型,并分析HPV DNA与E6/E7 mRNA检测的一致性。以47例病理诊断为金标准,计算并对比不同型别HPV DNA阳性者、不同型别HPV E6/E7 mRNA阳性者患高级别宫颈上皮内瘤变和宫颈癌的相对危险度(OR)。3、通过免疫组织化学方法,检测231例病理组织中叶酸受体、肿瘤细胞周期和表达相关蛋白的表达,并采用荧光定量PCR,检测宫颈脱落细胞中叶酸受体、肿瘤细胞周期和表达相关蛋白mRNA的表达；同时,向宫颈癌细胞株C33A中转染携带HPV E6、E7基因的质粒,通过Western Blot检测叶酸受体的表达变化。结果：1、高级别宫颈上皮内瘤变和宫颈癌发生的平均年龄为40.15±8.03岁。FRD诊断敏感度77.53%,特异度55.63%,与TCT相比无显著统计学差异。FRD诊断符合率64.07%,高于HPV和TCT,有显著统计学差异。FRD使用的小棉签对宫颈病变的诊断效率较低。与ASCUS时诊断TCT异常相比,≥ASCUS时诊断TCT异常的诊断效率更高。HPV检测对CIN2以上病变的阴性预测值为100%。HPV定量(HC2)与HPV分型诊断的敏感度和特异度无显著统计学差异。2、HPV16的感染率最高,其次是HPV52、HPV58。HPV E6/E7 mRNA检测的阳性率与HPV DNA拷贝数相关。高危型HPV混合感染和混合低危型HPV感染均不影响HPV E6/E7 mRNA分型检测。HPV 16、18 DNA阳性者发生CIN2以上病变的风险显著高于其他类型感染。HPV E6/E7 mRNA阳性者发生CIN2以上病变的风险显著高于单纯HPV DNA阳性者。HPV 16、18型E6/E7 mRNA阳性者,发生CIN2以上病变风险,显著高于HPV DNA阳性者和其他型HPV E6/E7 mRNA阳性者。3、叶酸受体、CREPT在组织病理学诊断的宫颈炎和低级别上皮内瘤变中均低表达,在高级别上皮内瘤变和宫颈癌中高表达,且有显著统计学差异。叶酸受体、CREPT mRNA在不同程度宫颈病变者的脱落细胞中无显著统计学差异。瞬时转染HPV E6、E7前后,C33A细胞中叶酸受体表达未见变化。结论：1、FRD诊断敏感度和特异度与TCT相比无显著统计学差异,且诊断符合率高于HPV和TCT。HC2与HPV分型诊断的敏感度和特异度无显著统计学差异,HPV检测对CIN2以上病变的阴性预测值极高。2、HPV E6/E7 mRNA阳性者发生CIN2以上病变的风险显著高于单纯HPV DNA阳性者。HPV 16、18型E6/E7 mRNA阳性者,发生CIN2以上病变风险,显著高于单纯HPV DNA阳性者和其他型HPV E6/E7 mRNA阳性者。3、叶酸受体、CREPT在组织病理学诊断的宫颈炎和低级别宫颈上皮内瘤变中低表达,在高级别宫颈上皮内瘤变和宫颈癌中高表达。叶酸受体、CREPT mRNA在不同程度宫颈病变者的脱落细胞中无显著统计学差异。瞬时转染HPV E6、E7前后,C33A细胞中叶酸受体表达未见变化。转染E7、共转染E6/E7后,CREPT的表达略有升高。
[Abstract]:Objective: To evaluate the application value of folic acid receptor mediated epithelial specific staining, tumor cell cycle and expression related protein and HPV E6/E7 mRNA typing in cervical cancer screening, and to clarify the expression of folic acid receptor, tumor cell cycle and expression related protein in cervical lesions and cervix exfoliative cells. Methods: 1, 231 outpatients For cervical cancer screening, prospective liquid based cytological examination (TCT), human papillomavirus detection (HPV) and folic acid receptor mediated epithelial special staining (FRD) examination, and pathological diagnosis as gold standard, were used to calculate and compare the sensitivity and specificity of TCT.HPV and FRD,.2, and 153 cases of cervical removal by fluorescence quantitative PCR. HPV DNA typing, HPV E6/E7 mRNA typing, and the consistency between HPV DNA and E6/E7 mRNA detection were performed. 47 cases of pathological diagnosis were the gold standard, and different types of HPV DNA positive were calculated and compared. The relative risk degree of high grade cervical intraepithelial neoplasia and uterine cervix cancer was affected by different types of HPV. The expression of folic acid receptor, tumor cell cycle and expression related protein in 231 cases of pathological tissue were detected, and fluorescence quantitative PCR was used to detect the expression of folic acid receptor, tumor cell cycle and expression related protein mRNA in cervical exfoliative cells; meanwhile, the plasmid carrying HPV E6, E7 gene was transfected into the cell C33A of cervix cancer, and the plasmid of E7 gene was passed through We Stern Blot detected the changes in the expression of folate receptor. Results: 1, the average age of high grade cervical intraepithelial neoplasia and cervical cancer was 40.15 + 8.03 years old.FRD diagnostic sensitivity 77.53%, specificity 55.63%, compared with TCT, there was no significant difference in.FRD diagnostic coincidence rate 64.07%, higher than HPV and TCT, there was significant statistical difference of.FRD used small cotton. The diagnostic efficiency of the cervical lesions was lower. Compared with the diagnosis of TCT abnormality when ASCUS, the diagnostic efficiency of the diagnosis of TCT abnormalities was higher than that of ASCUS. The negative predictive value of.HPV detection to the above CIN2 lesions was no significant difference between 100%.HPV quantitative (HC2) and HPV type diagnosis,.2, HPV16 infection rate was the highest, followed by HPV52. The positive rate of V58.HPV E6/E7 mRNA detection is related to the number of HPV DNA copies. High risk HPV mixed infection and mixed low risk HPV infection do not affect HPV E6/E7 mRNA type detection. The risk of.HPV 16,18 is significantly higher than that of other types of infection. .HPV 16,18 E6/E7 mRNA positive individuals with positive HPV DNA positive risk were significantly higher than those of HPV DNA positive and other HPV E6/E7 mRNA. The folate receptor was low in the pathological diagnosis of cervicitis and low grade intraepithelial neoplasia, and high level in high grade intraepithelial neoplasia and cervical cancer. There was no significant statistical difference between the folic acid receptor and CREPT mRNA in the exfoliated cells of different degree cervical lesions. Transient transfection of HPV E6, E7, and C33A cells did not change. Conclusion: 1, the sensitivity and specificity of FRD diagnosis are not significantly different from TCT, and the diagnostic coincidence rate is high. There was no significant difference in the sensitivity and specificity between the HPV and the TCT.HC2 and HPV types. The negative predictive value of the HPV detection was very high in the negative predictive value of the lesions above CIN2, and the risk of the CIN2 above the HPV E6/E7 mRNA positive was significantly higher than that of the simple HPV DNA. HPV DNA positive and other type HPV E6/E7 mRNA positive persons.3, folic acid receptor, CREPT in histopathological diagnosis of cervicitis and low grade cervical intraepithelial neoplasia, high expression in high level cervical intraepithelial neoplasia and cervical cancer. Folic acid receptor, CREPT mRNA in different degrees of cervical lesions of the exfoliated cells of no significant The expression of folate receptor in C33A cells did not change before and after transient transfection of HPV E6 and E7. After transfection of E7, the expression of CREPT increased slightly after transfection of E6/E7.
【学位授予单位】：中国人民解放军医学院
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R737.33

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