上皮性卵巢癌病因学研究进展

上皮性卵巢癌是当今最致命的妇科肿瘤之一,也是妇产科临床诊断及治疗中的最大难点。究其原因,除了早期缺乏明显的临床症状及有效的诊断方法外,还由于缺乏对上皮性卵巢癌发生、发展准确而全面的认识,故相关研究旨在更好地掌握上皮性卵巢癌的发病机制,有望在今后的诊断及治疗中取得新的进展。目前,上皮性卵巢癌公认的三大发病机制是：高促性腺激素理论、“二元论”和干细胞假说。本文对此作一综述。     

p53蛋白表达与上皮性卵巢癌恶性程度及预后的关系研究

目的：研究p53蛋白表达与上皮性卵巢癌恶性程度及预后的关系。方法：采用免疫组化SP法测定26例上皮性卵巢癌中p53蛋白表达。结果：p53蛋白在黏液性、浆液性、内膜样卵巢癌中的表达率（88.9%、75%、100%）无明显差异（P〉0.05）;三种卵巢癌的复发率分别为33.3%、12.5%、0,以黏液性癌最高,三者之间有显著差异（P〈0.05）;p53蛋白在淋巴结转移卵巢癌患者中均为高表达（100%）,与无淋巴结转移者（68.75%）相比差异明显（P〈0.05）;另外,淋巴结转移卵巢癌患者的复发率（40%）较无淋巴结转移患者（6.25%）明显增高（P〈0.05）。p53蛋白在I-II期和III-IV期卵巢癌的高表达（50%、100%）具有显著差异（P〈0.01）,不同期别卵巢癌复发的几率也存在显著差异,其中I-II期复发率为0,III-IV期复发率为31.25%。结论：p53蛋白的高表达与上皮性卵巢癌的恶性程度及预后有着密切的关系。     

ERK1/2通路基因突变对低级浆液性卵巢癌发生发展的影响

卵巢癌是女性生殖器官中常见的恶性肿瘤之一,是女性生殖系统肿瘤中的最大杀手.随着分子遗传学和肿瘤生物学研究的深入发展,基因诊断已成为治疗恶性肿瘤的重要手段.因此,研究卵巢癌的分子遗传学机制进而了解其生物学效应对卵巢癌的发生发展、治疗及预后等方面有重要意义.大量临床病理学和分子遗传学实验证明ERK1/2通路异常活动与低级浆液性卵巢癌的发生发展关系密切.文章将针对ERK1/2通路的异常持续活化(KRAS或BRAF突变次级效应)与低级浆液性卵巢癌发生发展的关联性研究作一简要综述.     

P53、C-erbB-2在上皮性卵巢癌中的表达及意义

目的:为了探讨C-erbB-2和P53在上皮性卵巢癌中的表达和意义及二者表达之间的关系,方法:应用免疫组织化学的方法回顾性研究了70例上皮性卵巢癌石蜡切片中这两个基因的蛋白表达情况,并对病人进行了生存期随访。结果:(1)70例上皮性卵巢癌中P53表达的阳性率为34.3％,P53在浆液性卵巢癌及粘液性卵巢癌间表达差异显著(P<0.05)。(2)C-erbB-2阳性率为31.47％。阳性细胞在瘤体局部具有“团块状”的特点。Ⅲ、Ⅳ期上皮性卵巢癌较Ⅰ、Ⅱ期显著为高(P<0.05)。(3)C-erbB-2和P53表达间无显著相关关系(P>0.05)。(4)Kaplan—Meier法研究表明,CerbB-2阳性患者生存期缩短(P<0.01),P53阳性者与阴性者无显著差异。Cox s比例风险回归模型研究表明,C-erbB-2表达状态、FIGO分期、残存瘤大小是三个对上皮性卵巢癌患者预后具有显著影响的变量。结论:C-erbB-2表达与上皮性卵巢癌的恶性程度关系密切。对其预后影响显著;而P53表达与其亚型有关。     

上皮源性卵巢癌干细胞的研究进展

肿瘤干细胞(cancer stem cells,CSCs)是肿瘤组织内具有自我更新能力以及无限增殖和多向分化潜能的一群细胞,对化疗药物耐受.卵巢癌干细胞(epithelial ovarian cancer stem cells,EOCSCs)在卵巢癌的发生发展、侵袭转移和耐药复发过程中都起到了重要作用.传统的肿瘤细胞减灭术联合顺铂、紫杉醇全身化疗对减小肿瘤体积,缓解临床症状具有一定的作用,但治疗后残留的EOCSCs能够短时间内重建肿瘤组织,是卵巢癌复发和难治的根本原因.深入了解EOCSCs的生物学特性,探索EOCSCs的发生发展机制,研究针对EOCSCs的靶向治疗药物,是抗肿瘤治疗的关键.     

PCNA、p53基因与上皮性卵巢癌预后的研究

目的探讨与上皮性卵巢癌预后有关的因子.方法回顾性分析89例上皮性卵巢癌的临床及病理资料,并用p53和PCNA单克隆抗体分别检测p53和PCNA的表达水平,用Cox模型对上述因子作生存分析.结果p53蛋白表达阳性率为62.9%,PCNA表达阳性率为82.0%.经单因素Cox模型分析,与预后有关的因子有淋巴转移、血道转移、直接蔓延、残存病灶、临床分期、血红蛋白值(P＜0.05);多因素分析,影响预后的独立因子有血道转移、临床分期、化疗疗程、PCNA定量(P＜0.05).结论临床分期、血道转移、PCNA定量和化疗疗程均是影响上皮性卵巢癌生存期的因素.     

上皮性卵巢癌52例分析

１９７６年至１９９２年，我院共收治卵巢上皮癌５２例，全部病例均采用手术为主配合化疗的综合疗法。其５年、１０年在活率分别为３３．３％及１７．４％。５年存活率Ⅰ、Ⅱ、Ⅲ、Ⅳ期分别为６６．７％、４４．４％、１６．６％、０％；粘液性囊腺癌６１．５％、浆液性囊腺癌３１．５％、宫内膜样癌及低分化癌均为２０．０％；无残余瘤６０．０％，残余瘤直径＜２厘米３７．５％，残余瘤直径＞２厘米６．３％；单一化疗１６．７％、联合化疗４７．７％。经统计学分析结果表明，临床分期、病理类型、残余瘤大小及化疗方案均为影响预后的因素。     

上皮性卵巢癌的危险因素分析

上皮性卵巢癌在卵巢恶性肿瘤中最常见。近年来，卵巢癌发病率有逐渐升高趋势。故本文采用病例对照研究的方法探讨生殖因素等与卵巢癌的关系，旨在寻找卵巢癌的高危因素，为预防卵巢癌的发生提供参考。资料与方法病例选择病例选自1994年3月一1995年3月住院的病人，所有病例均为经手术后病理学证实的上皮性卵巢癌，共127例。年龄为17岁一～3岁，平均年龄52岁。对照组选择选自同期住院的其他非恶性肿瘤病人共254例。无内分泌疾病和消化道疾病患者。对照组病人与卵巢癌病人的年龄相差不超过三岁。调查方法印制统一调查表格，调查人员均选自妇产…     

上皮性卵巢癌组织病理学及分子生物学研究进展

上皮性卵巢癌是最为常见的卵巢恶性肿瘤,根据组织病理学特征和分化程度可分为多种亚型.组织学类型和分化程度是决定肿瘤行为及预后的关键因素,了解不同亚型上皮性卵巢癌组织病理学和分子生物学特点,可为选择早期筛查的肿瘤标记物和制订针对性治疗方案提供参考.     

上皮性卵巢癌组织病理学及分子生物学研究进展

上皮性卵巢癌是最为常见的卵巢恶性肿瘤,根据组织病理学特征和分化程度可分为多种亚型.组织学类型和分化程度是决定肿瘤行为及预后的关键因素,了解不同亚型上皮性卵巢癌组织病理学和分子生物学特点,可为选择早期筛查的肿瘤标记物和制订针对性治疗方案提供参考.     

Clinical significance and prognostic relevance of KRAS,BRAF, PI3K and TP53 genetic mutation analysis for resectable and unresectable colorectal liver metastases: A systematic review of the current evidence

Background

Hepatic resection is considered the optimal potentially curative treatment for colorectal liver metastases (CRLM). Following resection, up to two-thirds of patients will develop recurrence within 5-years. Genetic mutation analysis of CRLM, especially KRAS status, has been proposed as a means to guide treatment, as well as identifying patients who can derive the most survival benefit from hepatic resection.

Analysis of KRAS,NRAS, BRAF,PIK3CA and TP53 mutations in a large prospective series of locally advanced rectal cancer patients

Little information is available on the clinical significance of cancer-related genes such as KRAS, NRAS, BRAF, PIK3CA and TP53 in nonmetastatic rectal cancer. We investigated mutations of these genes in a large prospective series of locally advanced rectal cancer (LARC) patients who were recruited into two phase II trials. Mutational analyses were performed with diagnostically validated methods including polymerase chain reaction, capillary electrophoresis single-strand conformational analysis, Sanger sequencing and next-generation sequencing. Associations between single or multiple gene mutations and clinicopathological characteristics and treatment outcomes were explored. Of these 269, 210 (78%) patients were assessable. Mutations of KRAS, NRAS, BRAF, PIK3CA and TP53 occurred in 43, 9, 4, 9 and 60% of patients, respectively. Concordance between paired biopsy and resection specimens was 82% for KRAS, 95% for NRAS, 99% for BRAF, 96% for PIK3CA and 63% for TP53. TP53 mutations were associated with extramural venous invasion on baseline MRI (78% vs. 65%, p = 0.04), poor pathological tumour regression (23% vs. 36%, p = 0.05) and a trend toward a worse 5-year progression-free survival (PFS; 60% vs. 74%, HR 1.59, p = 0.06). Patients with tumours harbouring mutation of TP53 and either KRAS or NRAS (32%) had a worse 5-year PFS than those with TP53/KRAS/NRAS wild-type tumours (54% vs. 72%, HR 1.75, p = 0.02). In univariate analysis, BRAF mutation predicted poor 5-year overall survival only among patients treated without cetuximab (20% vs. 73%, HR 3.29, p = 0.03). This is one of the largest biomarker studies in a prospective, largely homogeneous, LARC population. Our findings are hypothesis generating and require validation in independent series.     

新型人上皮性卵巢癌细胞HO8910靶向输送系统的构建及其生物学活性的鉴定

目的 构建新型人卵巢癌HO8910细胞株靶向输送系统TAT-OSBP-EGFP,并对其靶向输送特性和体外活性进行研究鉴定。方法 采用PGEX-6P-3质粒分别构建TAT-OSBP、OSBP-EGFP和TAT-OSBP-EGFP表达载体,重组质粒转化BL21（DE3）大肠杆菌,经IPTG诱导表达和GST SefinoseTM Resin柱亲和层析纯化后,采用SDS PAGE 和Western blotting 鉴定。流式细胞术分析不同浓度（0、1、5、10μmol/L）TAT-OSBP、OSBP-EGFP和TAT-OSBP-EGFP融合蛋白处理HO8910细胞2h后的细胞穿膜率,细胞免疫荧光法检测3种融合蛋白对HO8910的输送特性（以人结肠癌LoVo细胞作对比）,CCK-8法检测0、1、5、10、15、40、60、80、100μmol/L TAT-OSBP-EGFP处理HO8910细胞2h后的细胞活性。结果 成功构建TAT-OSBP-EGFP原核表达载体,并获得可溶性融合蛋白。与TAT-OSBP相比,当浓度为10μmol/L时,TAT-OSBP-EGFP处理后HO8910细胞的穿膜率无明显升高（P＞0.05）;但当浓度为1、5μmol/L时,TAT-OSBP-EGFP处理后HO8910细胞的穿膜率升高,差异有统计学意义（P＜0.01）。经TAT-OSBP-EGFP蛋白处理,HO8910细胞内荧光强度高于LoVo细胞;经TAT-OSBP蛋白处理,HO8910细胞内绿色荧光强度与LoVo细胞相似;经OSBP-EGFP蛋白处理,两种细胞内未见明显的绿色荧光。不同浓度TAT-OSBP-EGEP对HO8910细胞活性无影响（P＞0.05）。结论 成功构建针对人卵巢癌HO8910细胞株的靶向输送系统,为下一步肿瘤靶向药物输送载体的构建并发挥肿瘤的靶向杀灭作用打下了良好的基础。     

卵巢癌相关基因的筛选与鉴定

背景与目的:卵巢癌在妇科肿瘤中死亡率最高,但是目前对于在卵巢癌发生中的敏感和特异癌基因或抑癌基因的改变还知之甚少。因此,筛选新的卵巢癌相关基因并对其作用进行研究可能有助于探讨卵巢癌发生的机理,并可为卵巢癌的早期诊治提供有意义的生物学指标。本研究旨在获得新的卵巢癌相关基因。方法:在改良mRNA差异显示方法的基础上,结合反向Northern点杂交等鉴定方法,筛选卵巢癌组织和对侧正常卵巢组织之间的差异表达基因,进一步利用生物信息学资源对所获得的差异表达基因片段进行分析。并以差异表达片段作为探针,原位杂交方法检测基因在其它卵巢正常组织和卵巢癌组织中的表达。结果:筛选后得到12个差异表达基因,其中3个为基因组重复序列,5个未知基因序列,4个已知基因序列。原位杂交结果表明锌指蛋白361(Zincfingerprotein361,ZNF361)基因、蛋白酶体PSMA2(proteasomesubunit,alphatype2,PSMA2)基因、位于人1号染色体的未知基因OCRC13和位于9号染色体的未知基因OCRC4在36例卵巢癌组织中的表达均高于其在16例正常卵巢组织中的表达(P<0.05)。尤其是与鼠配子形成期特异表达家族成员-纺锤体蛋白(spindlin)基因高度同源的OCRC4基因,在16例正常卵巢癌组织中未见其表达,而在卵巢癌组织中的表达可达53%(17/36)。结     

Distinct molecular profiles in Lynch syndrome‐associated and sporadic ovarian carcinomas

Ovarian carcinoma in Lynch syndrome (LS) is associated with unexpectedly high survival; yet, beyond DNA mismatch repair (MMR) defects, the developmental mechanisms are unknown. We used established (genetic) and new (epigenetic) classifiers of ovarian cancer to explore similarities and differences between LS‐associated and sporadic diseases. To this end, all available ovarian carcinomas (n = 20) from MMR gene mutation carriers ascertained through a nation‐wide registry and 87 sporadic ovarian carcinomas of the main histological types were molecularly profiled. LS‐ovarian carcinomas were mostly of nonserous histology (12 endometrioid, seven clear cell and one serous), diagnosed at a mean age of 45.7 years, and associated with a 10‐year survival of 87%. Among LS‐ovarian carcinomas, 19/20 (95%) were MMR‐deficient vs. 11/87 (13%) among sporadic cases (p < 0.0001). In a striking contrast to the sporadic cases, the expression of p53 was normal and KRAS/BRAF mutations absent in all LS‐ovarian carcinomas. PIK3CA mutations, suggested to be a favorable prognostic factor, occurred with a frequency of 6/20 (30%), which was comparable to sporadic tumors of endometrioid or clear cell type. Tumor suppressor genes were more frequently methylated and LINE‐1 hypomethylation less common in LS‐ovarian carcinomas compared to their sporadic counterparts. The patterns of genetic and epigenetic alterations reflected the origin as LS vs. sporadic cases on one hand and the histological type on the other hand. In conclusion, the significant molecular differences observed between LS‐associated and sporadic ovarian carcinomas help explain the different behavior of these tumors and emphasize the need for tailored clinical management.     

p53 and related proteins in epithelial ovarian cancer

We conducted a retrospective immunohistochemical evaluation of the prognostic significance of the expression of p53 and the related proteins Bax, Bcl-2, growth arrest and DNA damage (Gadd45), murine double minute 2 (Mdm2) and p21^WAF1/CIP1 in chemonaïve tumours taken from 66 patients with ovarian cancer. Ki-67 expression (a marker of cell proliferation) was also evaluated immunohistochemically, while apoptosis within malignant cells was determined with the terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling (TUNEL) assay. The expression of each of the following proteins was significantly associated in the tumours (P<0.05 unless otherwise stated): Bax with Bcl-2 (P<0.01); Bax with Mdm2; p21^WAF1/CIP1 with Gadd45 (P<0.01); p21^WAF1/CIP1 with p53; p53 with Mdm2. Univariate analysis showed that expression of p53, Bax, bulk residual disease and International Federation of Gynecology and Obstetricians (FIGO) stage were all strongly correlated with response to chemotherapy (P<0.01). Similarly, the FIGO stage and Ki-67 expression (P<0.01), as well as pathological subtype and bulk residual disease (P<0.05), were prognostic factors for disease progression. The FIGO stage and Ki-67 expression were significant prognostic factors for overall survival (P<0.01), with Gadd45 expression and pathological subtype also significant (P<0.05) in a univariate analysis. Multivariate analysis for response to chemotherapy showed that expression of p53, Bax and FIGO stage were all independent prognostic factors (P<0.01). The FIGO stage was the most important independent prognostic factor for progression and survival on multivariate analysis (P<0.01). However, Ki-67 expression was also an independent prognostic factor for disease progression (P<0.05) and approached significance for survival (P=0.055). Taken together, these data suggest that determination of Ki-67 expression could supplement established prognostic factors.     

Alterations of theTP53 gene and TP53 protein expression in epithelial ovarian cancer before and after chemotherapy

Background A role for theTP53 (alias p53) tumor-suppressor gene in chemoresistance has recently been discussed, but little is known about the clinical relevance of theTP53 gene to chemoresistance. To elucidate the relevance ofTP53 status to chemoresistance, we investigated theTP53 gene and TP53 protein expression in tumors from the same patients, before and after chemotherapy. Methods Twenty-one patients with ovarian cancer, who had residual disease after primary surgery, were studied. These patients received chemotherapy consisting of cisplatin, doxorubicin, and cyclophosphamide, and then underwent a second surgery. Polymerase chain reaction-single strand conformation polymorphism analysis and cycle sequencing were performed to determineTP53 mutation. TP53 protein was detected by Western blot analysis. Results Of the 21 patients studied, 9 responded to chemotherapy. Mutation of theTP53 gene was seen in 7 patients (2 responders and 5 nonresponders) before chemotherapy. After chemotherapy, another mutation of the gene was observed in 5 patients, all of whom were nonresponders. TP53 protein was detected in 10 patients (3 responders and 7 nonresponders) before chemotherapy. After chemotherapy, the expression of TP53 protein increased in these 3 nonresponders, and became positive in 2 other nonresponders. Conclusions This study showed for the first time in clinical investigation that alterations toTP53 could develop in association with chemotherapy, and thatTP53 status may relate to the mechanisms of chemoresistance in patients with epithelial ovarian cancer.     

Impact of mutational status on survival in low-grade serous carcinoma of the ovary or peritoneum

Background:

Low-grade serous carcinoma of the ovary or peritoneum is a distinct, well- recognized histologic subtype characterized by young age at diagnosis, relative chemoresistance, and prolonged overall survival. Common mutations reported to be found within this subtype include KRAS and BRAF.

Methods:

Using clinical information of patients from our IRB-approved registry and tissue from a subset of these patients, we performed mutational analysis for KRAS and BRAF using the direct Sanger sequencing technique and correlated findings with the clinical outcome, overall survival (OS).

Results:

In 79 cases, patients with KRAS or BRAF mutations (n=21) had a significantly better OS than those with wild-type KRAS or BRAF (n=58) (106.7 months (95% CI, 50.6, 162.9) vs 66.8 months (95% CI, 43.6, 90.0)), respectively (P=0.018).

Conclusions:

Mutational status appears to be a potential prognostic factor in low-grade serous carcinoma of the ovary or peritoneum.     

卵巢癌肿瘤细胞减灭术中脾脏切除的手术探究

目的探讨晚期和复发的上皮性卵巢癌减瘤术中脾脏切除术的必要性和可行性。方法回顾分析首次（5例）或再次细胞减灭术（7例）中施行脾脏切除术的12例晚期卵巢癌患者的临床资料。结果12例患者的平均总生存期为37．75个月（15个月-65个月）,3年总生存率为58．33％（7／12）;7例生存至今的患者中2例无瘤生存,分别生存3年和3．5年,三年无瘤生存率为16．67％（2／12）。手术并发症发生率25％（3／12）,与脾切除直接相关的手术并发症发生率为8．33％（1／12）。获得满意减瘤术的9例患者和不满意减瘤术的3例患者的平均总生存期分别为43．33个月和21个月（t=3．215,P〈0.05）,有显著性差异。结论晚期或复发的卵巢癌肿瘤细胞减灭术时为达到满意的手术对脾脏转移者施行脾脏切除是必要和可行的。