【作者】 埃萨塔(Aissata Toure)；

【导师】 崔满华；

【作者基本信息】 吉林大学 ， 妇产科学， 2011， 博士

【摘要】 目的：在过去20年,多种肿瘤标志物在临床的广泛应用使肿瘤的诊断和治疗取得了很大的进展,尤其是在指导卵巢癌的临床治疗和早期预防方面具有重要的意义。近3年,随着实验室基础设施的改善和诊断方法的进步,通过比较各种早期诊断卵巢癌的诊疗方法的数据发现对CA125系列标记物检测的需求明显增加。本论文回顾性研究的目的是评估单独应用生物标志物癌抗原CA125准确、早期的发现中国及几内亚早期卵巢癌患者。方法：选择2005年3月1日-09年12月1日在中国吉林省长春市吉林大学白求恩医学部第二医院和西非几内亚科纳克里纳赛尔大学中央民族医院确诊为早期卵巢癌(Ⅰ-Ⅱ期)的患者度185例。应用生化ELISA技术,一种高分子量的糖蛋白,应用CA125试剂盒测定对照组和早期卵巢癌患者血清及手术切除标本中CA125值并对数据进行分析。我们应用酶联免疫吸附试验可以高效率的检测早期卵巢癌(Ⅰ-Ⅱ)患者外周血中CA125的水平。据文献报道检测早期卵巢癌患者外周血中CA125的水平已成为一个较优越的方法。统计分析方法：用SPSS15.0计算统计分析数据,评估患者的平均年龄(平均值±标准差)。P<0.05代表差异显著。本项研究围绕与CA125表达相关的关键因素展开,从而确定并规范诊断早期卵巢癌CA125的标准值。通过对早期卵巢癌患者外周血游离DNA的定性研究,探索可用于早期诊断、监测治疗卵巢癌的有效新方法。CA125被用于作为一种癌抗原,通过分析患者外周血肿瘤相关基因的表达以明确早期卵巢癌的诊断。结果：对无任何症状的早期卵巢癌患者,通过评估反应性抗原特异性的外周血CA125的值可有效诊断卵巢癌。检测外周血中卵巢细胞受抗原特异性早期反应产生的CA125可代表早期卵巢癌细胞内源性CA125的表达水平。本项研究证实在无症状的早期卵巢癌患者,卵巢癌细胞可引起CA125特异性地升高,从而可以进一步研究抗卵巢癌Ⅰ期和Ⅱ期的潜在抗体。因此,检测卵巢癌患者外周血中癌抗原CA125的表达可作为一个有前途的战略。有效的早期诊断无疑可以降低卵巢癌的死亡率,如果卵巢癌在出现临床表现多年前即被发现,如果对早期病变的有效诊断的准确率达80%或更高,则卵巢癌的死亡率将会减少一半。在227名早期卵巢癌患者中163名患者外周血中CA125表达呈阳性(>35 U/ml),二者相比其CA125的表达有差异(p<0.05)(患者年龄范围为18-77岁,中位数为49.89±14.96岁)。依据FIGO不同临床分期的患者CA125的表达水平是不同的。A-依据FIGO分期确定肿瘤分期：146例ⅡB期患者中64.31%的患者CA125表达呈阳性,其CA125的水平明显高于ⅠC和ⅡC的12例患者,CA125的表达与肿瘤的期别存在相关性。IB-IIB IC-IIB相比,P=4.375E-05,#IIC与IC-IIB相比P=0.013IB-IIB*与ⅠA相比,P=0.496,#IIC与ⅠA相比,P=0.431ⅠB～ⅡB与ⅠC-ⅡB相比,P=4.375E-05,#ⅡC与ⅠC-ⅡB相比,P=0.013,其差别有统计学意义Ⅰ期卵巢癌患者中22例CA125表达的水平升高,Ⅱ期卵巢癌患者中99例CA125表达水平升高,二者相比,Ⅱ期卵巢癌CA125表达的阳性率(11.95%)明显低于Ⅱ期卵巢癌阳性率(53.80%)B-早期卵巢癌和对照组检测227正常对照组,185例早期卵巢癌患者。其中185例早期卵巢患者中163例CA125的表达呈阳性,163例患者的血CA125呈高表达(>35 U/m1),12例CA125的表达呈阴性。163例早期卵巢癌患者与227例对照组相比,CA125的表达差异显著(P<0.05)。肿瘤>4cm的卵巢癌患者的外周血中肿瘤生物标志物CA-125的水平高于肿瘤<4cm的卵巢癌患者,但细胞浆内的CA125的水平与肿瘤的大小无显著关系。(0.019,P<0.05)。C CA125蛋白与早期卵巢癌患者临床病理特征的相关性。对于通过分析CA125表达阳性的卵巢癌患者,发现家庭妇女的卵巢癌患者的进展速度低于城市妇女。卵巢癌与其他肿瘤相比较,72.88%为单侧肿瘤,27.11%为双侧肿瘤。CA125表达水平与职业、年龄和教育水平无显著的统计学差异。18岁后结婚的患者的CA125表达阳性率(59.07%),分别要高于38-57岁结婚(57.62%)或18岁以前结婚的妇女(6.77%)。通过比较分析发现67.79%的已婚女性及76.27%的多产妇,CA125的表达也无显著差异。D-早期病理诊断的多样性卵巢癌患者CA125的表达比畸胎瘤,囊腺瘤,子宫内膜腺癌和非卵巢腺癌要高。进一步分析我们过去5年的研究的肿瘤类型的数据发现,5年内子宫内膜癌共14例(6.16%),2005-2007年6例(占42.85%).2008-2009年8例(占57.14%)。透明细胞癌4例,卵巢囊腺癌40例,浆液性卵巢癌26例,粘液性卵巢癌13例,卵巢布伦纳肿瘤和交界性卵巢肿瘤12例,卵巢腺癌5例,癌性腹水29例,卵巢良性畸胎瘤41例,卵巢恶性畸胎瘤20例,混合型25例。CA125表达水平在各肿瘤间表达无明显差异。2005年的病例数量(25.06%)低于2007年的病例数量(37.62%),最近3年收集病例157例。所有病例中,其中144例手术术后病理回报卵巢癌102例,不明来源的腺癌3例,良性卵巢肿瘤28例,子宫肌瘤8例,3例无异常。应用ELISA正态分布曲线区分正常组织和癌变组织样本的敏感性和特异性分别达到81.49%、71.80%。此外,通过敏感性和特异性的获得,从而可以区分卵巢癌的高危人群或低危人群。本次博士论文的结果：可考虑应用CA125的酶联免疫吸附试验作为一种发现早期卵巢癌患者的战略,从而降低妇女的癌症死亡率。结论：卵巢是盆腔内一个很小的女性生殖器官。卵巢通过分泌激素支持女性生殖系统。早期诊断卵巢癌可降低中晚期卵巢癌的高死亡率。早期卵巢癌患者的外周血中CA125呈刺激性的高表达。这个系统的功能能力需要进一步检测方能考虑临床的监测使用。更多还原

【Abstract】 BACKGROUND:Consultant gynecologic oncologists ELISA laboratory from the regional Register Cancer Center assisted community gynecologists’CA 125 test after diagnosis method for early ovarian cancer when they were invited. For this report, the study evaluated the performance CA125 effects of early stage on diagnosis outcome METHODS: The hospital files from 227 diagnosed for epithelial ovarian cancer between 2005-2009 of ConakryJilin2, ChinoGuineans, Africasia were abstracted. The diagnosis results were analyzed 4 for weak reaction ELISA; 154 for moderate; 27 for strong at the patients according to the clinicians examination by ELISA for diagnosis and SSP 15.00, Epi-Info6 analyses with univariate and multivariate Cox regression. RESULTS:Method was performed on 178 Bethune (BN) patients, and 7 Conakry (CY). ELISA Bethune followed diagnostically guidelines more strictly compared with Conakry,8CY(18.60%) vs.37BN(20.10%);[P=0.01]ⅠA-ⅠB stage; 8CY(18.20% vs. 30 BN(16.30%); [P=0.003]ⅡA-ⅡB stage,within 27 CY(62.79%) vs. 117 BN(63.58%); [P=0.02]ⅠC～ⅡC stage. Patients tumor were stratified according to early stageⅠwith 5-year overall survival rate was 96% versus 80%; (P=0.03) for patients disease and 76% versus 60%, (P=0.02) for stageⅡwho underwent peripheral blood ELISA CA125 Bethune and Conakry diagnosis. The hazards ratio was 0.79 (95% confidence interval [95%CI],0.61-1.03; adjusted for patient age, stage, type of hospital, when patients age 77 years were excluded, the hazards ratio fell to 0.71 (95% CI,0.54-0.94) in multivariate analysis. CONCLUSIONS:An early diagnosis ovarian cancers by ELISA CA125 occurred more often according to laboratory guidelines, tumor removal more often was complete, and survival improved.Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, Jilin University Medical Center Normal Clinical, Bethune, The ChinoGuineansCD 10183, ID 9200873101 ObjectiveIn the past 20 years the diagnosis have made a great effort a variety of tumor makers in clinical widely used. The guide clinical laboratory with early prevention has effective important ovarian cancer significance. The recent past 3 years showing an improvement of infrastructure and methods of diagnosis that there was an increased demand on CA125 assay over the past 5-years, particularly in the last 3 years compared of data variety early diagnosis method clinic of patients of early ovarian cancer. The retrospective study was designed performance as assessed by the area under the receiver biomarker cancer antigen CA125 alone of panel to correctly identify women significantly greater and performance with early ovarian cancer chinoguineans.MethodsThe biochemical ELISA technique, a high molecular weight glycoprotein, was measured in sera from patients with the most common cause of cancer death among gynecologic tumors stages (ⅠandⅡ) in the benign worldwide. For biomarker panel Bethunedonka (BD) in surgical resection specimens and control group, blood sample with early ovarian cancer clinical data underwent respective examination monoclonal CA-125 albumins with ovarian cancer diagnosed kit blood from March 2005 January 1 to 2009 December 31, the Second Jilin hospital, Clinical Bethune of Medical Science (BN) of International Exchange Center Jilin University, Changchun, Jilin, China and Registered Cancer Center, National Hospitals Donka, Gamal Abdel Nasser University, Conakry(CY), West Africa. Guinea.The ELISA data by In Vitro (protocol No.1820/ub90504A) kit sensitivity color reagent were purchased from a new biological Technology Co., Ltd, United States, and Alpha Diagnostic Intl.We observed high CA125 efficiencies in the early ovarian cancer (stageⅠ-Ⅱ) and in ELISA using low peripheral blood concentrations with the absence of significant cell viability impairment. Early ovarian cancer has evolved as a superior method for CA125 into peripheral blood cells, CA125 was expressed in 4 for weak reaction ELISA; 154 for moderate; and 27 for strong at the patients as reported in the literature.The expressing verified the results with the Gene Bank expression database analyzed cancer cells instability with this ELISA type, such as relevance of ovarian cancer stage I-II.Statistical analysis Methods:Two independent statisticians from program Epi-Info and SPSS15.0 calculate statistic analyzed the data; the age average (mean±SE) of patients will be evaluated. Significant difference was that p<0.05 which represented all probability values lower than 0.05. The study was performed encompassing the critical issues associated with expression efficiency, to standardize a CA125 protocol for use in early ovarian cancer ELISA schedules. Through the peripheral blood of patients with early ovarian cancer free DNA for qualitative research to explore new treatment can be used early diagnosis stage and efficacy of the method of monitoring. CA125 was used as an antigen cancer to optimize peripheral blood by early ovarian cells by diagnosis the expression of the reporter tumor gene analysis for confirmation of the optimized loading parameters.ResultsThe efficiency of this delivery system was assessed using CA125 peripheral blood in stimulating antigen specific peripheral blood cell responses in early ovarian stage of healthy individuals. The peripheral blood + antigen specific early ovarian cells responses were generated recognizing CA125 peripheral blood plasma and also early ovarian cancer cell lines endogenously expressing CA125. This study has identified CA125 specific early ovarian cells responses in healthy patients, allowing further investigation into the potential for its use as a stage I and II in ovarian cancer ELISA. Furthermore, the use of antigen cancer CA125 expressed with peripheral blood ovaries is a promising strategy for the delivery of CA125in the generation of antigen-specific early ovarian cell responses.Early diagnostic efficiency are undoubtedly the major priorities for achieving long-term reduction of mortality due to ovarian cancer the hypothesis that early ovarian cancer may be detectable up to many years prior clinical presentation and that if effective diagnosis for early stage disease were achieved with an accuracy of 80% or more, mortality would be halved. The positivity of serum concentrations of CA125 expression are elevated (i.e.>35 U/ml) in 163 blood of patients with early stage disease. The CA125 expressions in patients with varied FIGO clinical stage were different. The data case cohort (overall and by disease stage) median age (range) of the (n=227) was 49.89(18-77) years (mean±SE,49.89±14.96) of age on plasma biomarker concentration identified (Spearman’s rank correlation,0.0004. p< 0.05). A- Tumor type was identified and FIGO clinical stage by FIGO nomenclature Rio de Janeiro 1998, ACS Cancer Facts & Figures,2007 presented of early stage cohorts:⊥The CA125 expressions in patients with 146 cases (64.31%) IIB stage disease was much highest than in patients both with 12 (<35 U/ml) IC～IIC positive, the CA125 expression correlated the types, were significantly of IB to IIB, (0.496. p<0.05); and IIC (0.431,p<0.05).⊥IB-IIB compared to IC-IIB, P= 4.375E-05,# IIC compared to IC-IIB, P= 0,013⊥IB-IIB* compared to IA, P= 0,496,# IIC compared to IA, P= 0,431Patient’s commonly situation of early ovarian cancer, the significance difference was *IB～IIB compared to IC-IIB, P= 4.375E-05,# IIC compared to IC-IIB, P= 0,013.⊥The CA-125 expressions elevated (i.e.□35 U/ml) in 22 cases (11.95%) in patients with I stage, was lowest than in 99(53.80%) of II stage.B-The group Controls and early ovarian cancer⊥In biomarker (n= 185{43.59±13.12} Cases) and (n=227{49.89±14.96}, Cases). The positives (n=175{40.87±11.48}, group compared falsies positives (n=6{27.09±10.14}). The significant differences biomarker plasma concentrations were significantly greater within 154(4CY+150BN) with 38.60±15.12 average (0.44, p< 0.05) moderate,27(1CY+26BN), 35.08±10.08 for strong,4 (2 CY and 2 BN)= 17.03±5.08 for weak reaction at patients and early ovarian cancer positively of date in Clinical patients 224(0.0003, p<0.05) of CA125 alone.⊥High levels (<35 U/ ml) were found in 4 cases of 227 patients with active ovarian cancer but in 181 (79.73 %) Bethunedonka (BD) patients who had positive of cases.⊥The tumor size on biomarker CA-125 plasma concentration was highly than in cells tumor>2 cm (177.24%) i.e 97.62% of the ovarian cells tumor<2 cm, plasma concentration was no significantly with tumor size, (0.019,p<0.05)⊥Anatomical seat 18-37 ages were usually much affected,63 cases i.e.7 cases tumors associated, (0.299, p<0.05). Histological diagnosis by age and anatomical areas (0.299, p<0.05) compared to anatomical areas by age (0.0006, P<0.05).C- Correlation between CA-125 protein and clinicopathological features of the patients of early ovarian cancer.⊥The housewives are the most affected layer less than urban women to increase with processing,62.71%, with 142.37% of cases respectively. Compared with ovarian tumors unilateral or bilateral with other tumor,72.88%,27.11% of the cases registered. There were no statistical significantly difference correlates in profession (0.032, p<0.05) and age> 58 old (0,286, p<0.05) compared to education level (0.009, p< 0.05) with CA125A cancer antigen positively. CA125 positive expressing was seen in 17 of patients who get married before 18 old and 146(78.91%) after 18 years old; high than the age group 38 to 57 57.62%, and the age group below 18 years represents 6.77% respectively. Compared to married,67.79% and multiparous,76.27%patients. The CA125 expression were also no statistical significantly difference inthe parity (0.073, p<0.05), complaints (p=0.091, p<0.05) and in age marriage (>18 old years,0.106,p<0.05) for cohorts.D-According to the variety histopathological diagnosis clinic in early stage4-CA125 was expressing much than the teratoma (417.62%) with cystadenocarcinome, endometrial adenocarcinoma, and non ovarian adenocarcinomas.4 In five (5) past years, the data in our study were further analyzed with types and detected in 14(6.16%) cases with endometrioid carcinoma with 5-year relative diagnosis estimates increasing from 6(42.85%) in 2005-2007 to 8(57.14%) in 2008-2009.⊥We in 4(2.12%) clear cell carcinoma were diagnosed. The significant difference was in 2005, (0.03, P<0.05) and 2007, (0.02 p<0.05) diagnosis. In cystadenocarcinoma (n=40), serous,(n=26), mucinous(n=13), Brenner tumor and borderline(n=12), Adenocarcinoma(n 5), Leiomyoma et ascite carcinomatous(n =29). Cysts and teratoma tumors(n = 41) with carcinoma(n = 20), mixed type (mixed, n = 25). Tumor typing was no significantly with CA125.⊥The greatest frequency was observed in 2005 which is lower than that in 2007 diagnosis 25.06% vs. (37.62%), respectively. Almost,157(145.92%) cases were detected in the recent past 3 years.⊥Of those patients,144 cases underwent surgical procedures.5 had ovarian cancers,3 had adenocarcinoma of unknown origin,28 had benign tumors,8 had fibroids, and 3 had no abnormalities.Using ELISA normal distribution statue to classify the measurements a sensitivity of 81.49 % and 71.80% specificity was achieved when classifying normal and cancerous samples. Also, a sensitivity of specificity was obtained when classifying patients as either being in a high risk group or low risk group of developing ovarian cancer. The results gained from this PhD thesis should be taken into consideration when designing future CA125 ELISA strategies, to combat one of the leading causes of cancer mortality in women, early ovarian cancer.ConclusionThe ovaries are small female reproductive organs that reside in the pelvis. The ovary makes female hormones reproduction system. Diagnosis of disease in early stages can reduce the high mortality rate of ovarian cancer and treat of disease in advanced stages. CA125 is not a novel strategy to express peripheral blood and a strong expression simultaneously in ovaries cells earlier. The functional capacity of this system requires further monitoring before it can be considered for investigation clinical use.更多还原