细胞周期因子Geminin、Cdt1、P16及Ki67在宫颈上皮内瘤变组织中的差异表达研究

发布时间：2018-01-25 05:20

本文关键词： 宫颈上皮内瘤变 Geminin Cdt1 mRNA 宫颈上皮内瘤变 Geminin P16 Ki67 HPV 宫颈上皮内瘤变宫颈锥切术全子宫切除术阴道残端　出处：《南京医科大学》2017年博士论文　论文类型：学位论文

【摘要】：第一部分细胞周期因子Geminin与Cdt1的mRNA在宫颈上皮内瘤变组织中的表达研究目的:探讨细胞周期因子Geminin与Cdt1的mRNA在宫颈癌前病变的发生、发展中的作用,为宫颈癌前病变的筛查、随访及预后评估提供新的生物学指标。方法:宫颈病变患者共210例为研究对象(CINⅠ、Ⅱ、Ⅲ组织各70例),阴道镜下病变周围区域组织病理证实为粘膜慢性炎55例作为宫颈炎症组。因子宫肌瘤或子宫腺肌症住院行全子宫切除术的宫颈组织30例为正常对照组,术前TCT、HPV检查均阴性,年龄22-65岁,平均38± 12.5岁,宫颈标本均经病理证实明确诊断,且均未接受过化疗、免疫及放射等治疗,取材前均征得患者知情同意,此研究获得江苏省人民医院伦理委员会批准。采用RT-PCR法检测宫颈正常组,宫颈炎症组,CINⅠ组、CINⅡ及CINⅢ组中Geminin及Cdt1 mRNA的表达量。结果:GemininmRNA在正常宫颈组织、宫颈炎症、CINⅠ级、CINⅡ级、CINⅢ级五组中表达量分别为 0.56±0.00、0.48±0.10、2.23±0.23、3.48±0.54、5.07±0.28。Cdt1 mRNA 表达量分别为:1.11±0.00、1.01±0.58、2.87±1.24、4.09± 1.64、5.52± 1.36。随着CIN等级的增高,Geminin及Cdt1 mRNA的表达逐渐升高,组间比较及与正常组、宫颈炎症组比较差异均有统计学意义(p0.05),正常宫颈与宫颈炎症组比较无统计学意义(p0.05)。结论:细胞周期因子Geminin与Cdt1 mRNA在宫颈病变组织中表达异常,并与病变程度呈正相关,说明HPV感染后通过改变细胞周期因子的表达而干扰宫颈上皮细胞增生周期,进而发生上皮内瘤变甚至癌变,为进一步研究HPV致癌的发病机制提供帮助,为临床尽早明确诊断、预测宫颈病变提供可能的生物学参数。第二部分Geminin与p16、Ki67在宫颈上皮内瘤变组织的差异表达及与HPV关系的研究目的:探讨细胞周期因子Geminin与p16、Ki67在宫颈病变组织的差异表达,比较Geminin与p16、Ki67在宫颈病变中的诊断及预测价值,及其与HPV感染的相关性。方法:宫颈病变患者共210例为研究对象(CINⅠ、Ⅱ、Ⅲ组织各70例),阴道镜下病变周围区域组织病理证实为粘膜慢性炎55例作为宫颈炎症组。因子宫肌瘤或子宫腺肌症住院行全子宫切除术的宫颈组织30例为正常对照组,术前TCT、HPV检查均阴性,年龄22-65岁,平均38±12.5岁,宫颈标本均经病理证实明确诊断,且均未接受过化疗、免疫及放射等治疗,取材前均征得患者知情同意,此研究获得江苏省人民医院伦理委员会批准。采用免疫组化方法法检测宫颈正常,宫颈炎症,CINⅠ、CIN及CINⅢ组织中Geminin、p16及Ki67的差异表达、采用HC2的方法检测HPV的载量。结果:Geminin、P16在正常宫颈及宫颈炎组织中无阳性或弱阳性表达。Geminin在 CINⅠ、CINⅡ、CINⅢ 中的阳性表达分别为 12.9%、54.3%、92.9%(p0.05);P16的阳性率为15.7%、61.4%、87.1%(p0.05)。Ki67在正常宫颈及宫颈炎组织中分别有3.3%及5.5%阳性表达,在CINⅠ、CINⅡ、CINⅢ中有35.7%、82.9%、95.7%阳性表达(p0.05)。随着CIN等级的增高,Geminin及p16、Ki67的表达逐渐升高,组间比较及与正常组、宫颈炎症组比较差异均有统计学意义(p0.05),正常宫颈与宫颈炎症组比较无统计学意义(p0.05)。HPV在宫颈炎、CINⅠ、CINⅡ、CINⅢ 组织中位数分别53.2、118.2、354.6、489.7(p0.05)。Geminin 的特异度及阳性预测值最高。Geminin、Ki67的异常表达与HPV感染有相关性。结论:Geminin对宫颈上皮内瘤变程度的灵敏度略低于P16、Ki67,但特异度、阳性预测值均高于P16、Ki67,而阴性预测值及精确度介于P16与Ki67之间,Geminin及Ki67的表达与HPV的感染呈正相关,而p16的表达与HPV的感染无相关性。提示Geminin在临床对宫颈病变诊断方面更有优越性,在宫颈病变组织中可考虑作为新的细胞周期因子指标来协助早期、精确的诊断和预测宫颈病变。第三部分宫颈高级别瘤变行全子宫切除术相关临床病理参数探讨目的:分析宫颈高级别瘤变患者行全子宫切除术,术前宫颈细胞学的分布、HPV的载量、宫颈活检组织病理与阴道镜、宫颈锥切的符合率,及宫颈锥切切缘阳性与病灶残留的相关性,以及术后阴道残端病变发生的风险,为临床评估CINⅡ-Ⅲ患者全子宫切除的必要性及阴道残端随访提供依据。方法:收集了 368例因CINⅡ-Ⅲ行全子宫切除的患者,分别从宫颈细胞学、HPV与宫颈病变的相关性、阴道镜判断与宫颈活检病理的符合率、宫颈锥切术后切缘阳性与全子宫切除术后病理结果的对比、阴道残端日后随访情况等多方面回顾分析。结果:所有患者TCT检查结果中ASCUS的比例最高(40.5%),但不同细胞学结果之间HPV的载量之间差异无统计学意义(p0.05)。宫颈锥切术切缘阳性的病例,与全子宫切除术后病灶残留有直接相关性(p0.05)。阴道残端发生VAIN的比例为11.1%符合发病率。结论:宫颈高级别瘤变患者行全子宫切除术需个体化处理,对阴道残端的随访不可忽视。
[Abstract]:Objective to study expression of Geminin and Cdt1 mRNA of the first part of the cell cycle factor in cervical intraepithelial neoplasia: a study of Geminin and Cdt1 cell cycle factor mRNA in cervical precancerous lesions, the effect of the development, for the screening of cervical precancerous lesions, provide a new biological marker to evaluate prognosis. Patients with cervical lesions: a total of 210 cases as the research object (CIN I, II, III group 70 cases each), colposcopy and pathology for regional organizations around the lesion mucosa of 55 cases of chronic inflammation as the inflammation group. Because of uterine myoma or adenomyosis hospital received hysterectomy for 30 cases of normal cervical tissue the control group, preoperative TCT, HPV were negative, 22-65 years old, average 38 + 12.5 years, cervical specimens were pathologically confirmed diagnosis, and had not received chemotherapy, immune and radiation therapy, patients were drawn before the consent of the informed consent. Study on the approval of the ethics committee of Jiangsu Province People's Hospital. RT-PCR method was used to detect cervical inflammation group, normal group, CIN group, the expression of CIN II and CIN in Geminin and Cdt1 group mRNA. Results: GemininmRNA in normal cervical tissues, cervical inflammation, CIN grade, CIN grade, CIN grade III expression in the five groups the amount was 0.56 + 0.00,0.48 + 0.10,2.23 + 0.23,3.48 + 0.54,5.07 + 0.28.Cdt1 mRNA expression respectively: 1.11 + 0.00,1.01 + 0.58,2.87 + 1.24,4.09 + 1.64,5.52 + 1.36. with the increase of CIN level, the expression of Geminin and Cdt1 of mRNA increased gradually, and the comparison between groups with the normal group, the cervicitis group had significant difference (P0.05), normal cervical and cervicitis groups were not statistically significant (P0.05). Conclusion: the abnormal expression of cyclin Geminin and Cdt1 mRNA in cervical lesions, and correlated with the lesion degree of positive, said HPV after infection by altering the expression of cell cycle factor and interference of cervical epithelial cell proliferation cycle, and thus the occurrence of intraepithelial neoplasia and cancer, to provide help for the further research of the pathogenesis of HPV cancer, clinical early diagnosis and prediction of cervical lesions may provide biological parameters. In the second part, Geminin and p16, to study differential expression Ki67 in cervical intraepithelial neoplasia and its relationship with HPV: To investigate the Geminin and p16 cell cycle factor, differential expression of Ki67 in cervical lesions, Geminin and p16, the value of Ki67 diagnosis and prediction in cervical lesions, and their correlation with HPV infection. Methods: a total of 210 cases of patients with cervical lesions the object of study (CIN I, II, III group 70 cases each), colposcopy and pathology for regional organizations around the lesion mucosa of 55 cases of chronic inflammation as the inflammation group. Because of myoma of uterus or uterine gland Muscle disease who underwent hysterectomy for cervical tissues in 30 cases of normal control group, preoperative TCT, HPV were negative, 22-65 years old, average 38 + 12.5 years, cervical specimens were pathologically confirmed diagnosis, and had not received chemotherapy, immune and radiation therapy, were drawn with the consent of the patients informed consent, approved by the research ethics committee of Jiangsu Province People's Hospital. Using immunohistochemistry method in normal cervical, cervicitis, CIN I, Geminin CIN and CIN in tissues. The differential expression of p16 and Ki67, load using HC2 method to detect HPV. Results: Geminin, P16 positive or weakly positive expression of.Geminin in CIN I and CIN II in normal cervix and cervicitis tissues, the positive expression in CIN were 12.9%, 54.3%, 92.9% (P0.05); the positive rate of P16 was 15.7%, 61.4%, 87.1% (P0.05).Ki67 in normal cervix and cervicitis tissues were 3.3% and 5.5 % positive expression in CIN I, CIN II, CIN III in 35.7%, 82.9%, 95.7% positive expression (P0.05). With the increased levels of CIN, Geminin and p16, Ki67 expression gradually increased, compared with the normal group and the inflammation group, there were significant differences (P0.05), normal cervical and the inflammation group was not statistically significant (P0.05).HPV in cervicitis, CIN I, CIN II, CIN III tissues were 53.2118.2354.6489.7 median (P0.05).Geminin specificity and positive predictive value of the highest.Geminin, HPV expression and Ki67 infection correlation. Conclusion: the sensitivity of Geminin to the degree of cervical intraepithelial neoplasia the slightly lower than P16, Ki67, but the specificity, the positive predictive value was higher than that of P16, Ki67, and the negative predictive value and accuracy between P16 and Ki67, HPV expression and Geminin infection was positively related to Ki67 and the expression of HPV and p16, and the infection had no correlation. Geminin in the clinical diagnosis of cervical lesions more superiority in cervical lesions can be considered as a new index of cell cycle factor to assist early, accurate diagnosis and prediction of cervical lesions. The third part of the cervical high-grade intraepithelial neoplasia hysterectomy clinical pathological parameters Objective: analysis of cervical high-grade intraepithelial neoplasia patients with hysterectomy, distribution of cervical cytology before operation, load HPV, cervical biopsy and the coincidence rate of colposcopy, cervical conization, and the relationship between cervical lesions and early cone positive margin residual, risk of vaginal stump lesions in and after operation, provide the basis for clinical evaluation of CIN II in patients with hysterectomy and vaginal stump need follow-up. Methods 368 cases with CIN II - III patients with hysterectomy were, respectively from the correlation of cervical cytology, HPV and cervical lesions, Yin The coincidence rate of tract judgment and cervical biopsy, comparison of cervical conization margin positive and hysterectomy pathology, were analyzed in the aspects of vaginal stump after review. Results: all patients TCT examination results in the highest proportion of ASCUS (40.5%), but the difference was not statistically significant between the load between different cytology results of HPV (P0.05). Cervical conization margin positive cases, there is a direct correlation with residual lesions after hysterectomy (P0.05). The vaginal stump VAIN ratio was 11.1% with the incidence. Conclusion: the cervical high-grade intraepithelial neoplasia patients with hysterectomy need to be individualized treatment of vaginal stump up can not be ignored.

【学位授予单位】：南京医科大学
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R737.33

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