卵巢黏液性肿瘤中EGFR表达和K-RAS基因的突变

目的 研究卵巢黏液性肿瘤中表皮生长因子受体(epidermal growth factor receptor,EGFR)蛋白的表达及K-RAS基因的突变,探讨卵巢黏液性肿瘤的发病机制及靶基因治疗的可行性.方法 应用免疫组化PV 9000两步法和PCR-RFLP法分别检测20例卵巢黏液性囊腺瘤、40例卵巢黏液性交界性囊腺瘤和40例卵巢黏液性囊腺癌中EGFR蛋白的表达和K-RAS基因的突变情况.结果 EGFR在卵巢黏液性囊腺瘤、黏液性交界性囊腺瘤和囊腺癌中的阳性表达率分别为0、37.5%、67.5%(P<0.01).K-RAS在卵巢黏液性囊腺瘤、黏液性交界性囊腺瘤和囊腺癌中的突变率分别为0、37.5%、7.5%,交界组突变率明显高于其他2组,但卵巢囊腺瘤与囊腺癌比较,差异无统计学意义(P>0.05).EGFR与卵巢黏液性囊腺癌的临床分期、病理分级、患者年龄无关(P<0.05),与肿瘤的大小相关(P<0.05).EGFR与卵巢黏液性交界性肿瘤临床病理分期、肿瘤大小、患者年龄无关(P>0.05).K-RAS基因突变与卵巢黏液性交界性肿瘤临床病理分期、肿瘤大小无关(P>0.05),与患者年龄相关(P<0.05).EGFR的表达和K-RAS基因的突变在卵巢黏液性交界性肿瘤中无相关性(P>0.05).结论 EGFR对卵巢黏液性交界性囊腺瘤和卵巢黏液性囊腺癌的形成起到了一定的作用,而K-RAS基因则可能是卵巢黏液性交界性囊腺瘤形成的一个重要因素.     

卵巢黏液性交界性肿瘤中K-ras基因突变及p21 ras蛋白的表达

目的：观察卵巢黏液性交界性肿瘤中K-ras基因突变及p21 ras蛋白的表达,探讨卵巢黏液性交界性肿瘤的发病机制及靶基因治疗的可能。方法采用PCR-RFLP法和免疫组化EliVision法分别检测40例卵巢黏液性交界性囊腺瘤、40例卵巢黏液性囊腺癌和20例卵巢黏液性囊腺瘤中K-ras基因突变和p21 ras蛋白的表达。结果 K-ras基因在卵巢黏液性囊腺瘤、黏液性交界性囊腺瘤和黏液性囊腺癌中的突变率分别为0、37.5%、7.5%,交界组突变率明显高于其他两组,差异有统计学意义( P<0.01)。 K-ras基因突变在卵巢黏液性交界性肿瘤年龄分组中突变,差异有统计学意义(P<0.05)。 p21ras蛋白在卵巢黏液性囊腺瘤、黏液性交界性囊腺瘤和黏液性囊腺癌中阳性率分别为5%、45%、10%,交界组阳性率明显高于其他两组,差异有统计学意义(P <0.01)。结论 K-ras基因突变及p21ras蛋白表达可能是卵巢黏液性交界性肿瘤形成的原因之一,有利于卵巢黏液性交界性肿瘤的判断,还可为临床作为靶向治疗药物分析提供病理学基础。     

HDAC1和HDAC6蛋白过表达在卵巢浆液性癌中的临床病理学意义

目的探讨组蛋白去乙酰化酶(histone deacetylase,HDAC)1和6蛋白表达检测在卵巢浆液性癌中的临床病理学意义。方法选取147例卵巢病变的存档蜡块标本,其中包括39例卵巢浆液性囊腺瘤26例卵巢浆液性交界性囊腺瘤和82例卵巢浆液性囊腺癌。应用免疫组化EnVision法检测HDAC1和HDAC6基因蛋白在上述卵巢病变组织中的表达水平,并分析其蛋白检测的临床病理学意义。结果 HDAC1和HDAC6蛋白分别为胞核和胞质染色,HDAC1蛋白强阳性表达率在卵巢浆液性囊腺瘤、交界性浆液性肿瘤和浆液性囊腺癌中分别为7.7%(3/39)、65.4%(17/26)和80.5%(66/82);HDAC6蛋白的强阳性表达在39例卵巢浆液性囊腺瘤中完全阴性,而在交界性浆液性肿瘤中为34.6%(9/26)、在浆液性囊腺癌中则高达86.6%(71/86),差异均具有统计学意义。结论 HDAC1和HDAC6蛋白在卵巢癌发生发展过程中起重要作用,对于卵巢浆液性癌的诊断具有重要的辅助意义,有望成为卵巢癌治疗的新靶点。     

NQO1过表达在卵巢黏液性囊腺癌预后评估中的意义

目的:探讨NAD(P)H醌氧化还原酶1[NAD(P)H-quinone oxidoreductase 1,NQO1]过表达在卵巢黏液性囊腺癌临床预后评估中的意义。方法:应用免疫组化En Vision法检测NQO1蛋白在162例卵巢黏液性囊腺癌组织、35例卵巢黏液性囊腺瘤组织和29例正常卵巢上皮组织中的表达,并分析其过表达与卵巢黏液性囊腺癌临床病理学特点之间的关系,通过Kaplan-Meier方法进行生存分析。结果:NQO1蛋白在卵巢黏液性囊腺癌组织中的阳性率及强阳性率分别为85.8%和64.2%,显著高于卵巢黏液性囊腺瘤和正常卵巢上皮组织(P0.01)。卡方检验结果显示,NQO1蛋白高表达与卵巢黏液性囊腺癌组织学分级和FIGO分期密切相关(P0.05)。Kaplan-Meier生存分析显示,NQO1蛋白高表达的卵巢黏液性囊腺癌患者总生存期和无病生存期均明显低于NQO1蛋白低表达患者。结论:NQO1蛋白在卵巢黏液性囊腺癌组织中呈高表达,可能成为卵巢黏液性囊腺癌预后评估的有效生物学指标。     

卵巢上皮性癌中inhibin及其受体β-glycan的表达

目的 探讨α-inhibin及其受体β-glycan在恶性卵巢上皮性肿瘤中的表达,评估两种多肽信号对卵巢癌生长的调控作用,探讨卵巢癌的发病机制.方法 应用EliVision方法检测30例正常卵巢组织、30例卵巢囊腺瘤和80例卵巢囊腺癌中α-inhibin及其受体β-glycan的表达.结果 α-inhibin在正常卵巢、卵巢囊腺瘤和卵巢囊腺癌中的总阳性表达率分别为93.3%、60.0%、36.3%,差异均有显著性(P<0.01);β-glycan在正常卵巢、卵巢囊腺瘤、卵巢囊腺癌的阳性表达率呈逐渐下降的趋势分别为100%、80.0%、75.0%,卵巢囊腺瘤、囊腺癌与正常卵巢组比较,差异有显著性(P<0.05),但卵巢囊腺瘤与囊腺癌比较,差异无显著性(P>0.05).结论 α-inhibin蛋白在卵巢囊腺癌中表达明显下降,而β-glycan在卵巢肿瘤中的表达仍然很高,提示β-glycan对增强α-inhibin抑制卵巢囊腺癌的形成可能起着一定的作用.     

卵巢浆液性交界性肿瘤中clusterin的表达

目的探讨凋亡抑制因子clusterin在卵巢浆液性肿瘤中发生、发展的作用及其与bcl-2、Ki-67表达的关系。方法免疫组化sP法检测clusterin、bcl-2、Ki-67在20例卵巢浆液性囊腺瘤,28例浆液性交界性肿瘤,26例浆液性囊腺癌标本中的表达。结果clusterin在囊腺瘤、交界性肿瘤、囊腺癌中的阳性率分别为40％,67．9％,96．2％。囊腺癌中的clusterin与Ki-67的表达水平均明显高于囊腺瘤（P〈0．05）和SBOT（P〈0．05）。交界性肿瘤中有腹水的患者clusterin阳性率要明显高于无腹水者（P=0．0390）,Ki-67在有腹膜种植的患者阳性率要明显高于无腹膜种植者（P=0．0473）。且两者的表达成正相关。结论clusterin基因可能通过抑制凋亡在卵巢浆液性肿瘤的发生发展中起重要作用。clusterin与Ki-67结合起来分析可能作为鉴别SBOT与浆液性癌的辅助指标。     

Survivin和Smac在卵巢黏液性肿瘤中的表达及意义

目的 探讨Survivin和Smae在卵巢黏液性肿瘤中的表达及意义.方法 应用免疫组织化学SABC法检测55例原发性卵巢黏液性肿瘤中的Survivin和Smac表达,并应用免疫电镜胶体金标记法观察Smac在原发性卵巢黏液性肿瘤中的亚细胞定位及表达趋势.结果 (1)Survivin在良性、交界性和恶性组的阳性表达率分别为2/20、12/15和20/20,呈升高趋势,其中良性与交界性、良性与恶性组间比较差异均有统计学意义(P<0.01).良性、交界性和恶性组Survivin的胞核阳性表达率分别为1/20、6/15和5/20,其中良性与交界性组比较差异有统计学意义(P<0.05).Smae在良性、交界性和恶性组的阳性表达率分别为19/20、9/15和3/20,呈降低趋势,三组间两两比较差异均有统计学意义(P<0.01或<0.05).Survivin和Smac之间呈负相关(r=-0.153,P<0.01).(2)良性、交界性和恶性组线粒体内Smae金标颗粒计数分别为24.1±7.2、11.1±1.9和5.2±1.7,三组间两两比较差异均有统计学意义(P<0.01或<0.05);线粒体外计数分别为4.7±3.0、2.9±1.0和1.7±1.3,三组间比较差异无统计学意义(P>0.05).结论 随着卵巢黏液性肿瘤的恶性进展,凋亡抑制蛋白Survivin表达上调,促凋亡蛋白Smac表达下调;下调的Smac蛋白表达主要为储存在线粒体膜间的非活性形式蛋白,而非释放到胞质中的活性形式蛋白.     

Numb蛋白在卵巢浆液性肿瘤中的表达及临床意义

目的检测Numb蛋白在卵巢浆液性肿瘤组织中的表达及其临床意义。方法用免疫组织化方法检测Numb蛋白在卵巢浆液性囊腺癌、交界性浆液性囊腺瘤、浆液性囊腺瘤及正常卵巢组织中的表达情况,并与患者的临床病理资料进行分析。结果Numb蛋白在卵巢浆液性囊腺癌中表达阳性率明显低于卵巢正常组织和卵巢浆液性囊腺瘤及交界性浆液性囊腺瘤的阳性率(P0.01),但与肿瘤分化程度、临床病理分期、淋巴结转移及年龄均无明显相关性(P0.05)。结论Numb蛋白在卵巢浆液性囊腺癌中低表达,可能参与浆液性囊腺癌的发生过程。     

卵巢浆液性肿瘤中IQGAP1与E-cadherin表达的检测

目的探讨上皮性钙黏素(E-cadherin)与具有IQ结构域的人Ras GTP激活蛋白相关蛋白1(IQGAP1)在卵巢浆液性肿瘤中的表达及其临床意义。方法分别用免疫组织化学SP法和Western blot法检测20例卵巢浆液性囊腺癌、10例卵巢交界性浆液性囊腺瘤、10例卵巢浆液性囊腺瘤以及10例正常卵巢组织中E-cadherin和IQGAP1的蛋白表达。结果 E-cadherin蛋白在浆液性囊腺瘤中表达最高,明显高于正常卵巢组织和浆液性囊腺癌组织(P0.05),其均值高于交界性囊腺癌,无统计学意义。IQGAP1在卵巢浆液性囊腺癌中表达较正常卵巢组织、良性及交界性肿瘤组织均增高(P0.05);免疫组织化学染色证实IQGAP1在浆液性囊腺癌中以胞膜表达为主,而在良性肿瘤中以胞质表达为主。结论 E-cadherin和IQGAP1在卵巢浆液性瘤高表达,可联合用于卵巢浆液性肿瘤的免疫组化诊断。     

ataxin-3在卵巢浆液性肿瘤中的表达及其临床意义

目的检测ataxin-3在卵巢浆液性肿瘤中的表达及临床意义。方法采用免疫组化SP法检测ataxin-3在各组卵巢浆液性肿瘤中表达情况。结果ataxin-3在卵巢低级别浆液性囊腺癌中表达水平明显高于卵巢浆液性囊腺瘤、交界性浆液性囊腺瘤、高级别浆液性囊腺癌中的表达水平,但后三者之间无明显差异(P0.05),ataxin-3表达与卵巢浆液性囊腺癌的分化程度呈负相关,与其它临床病理因素均不存在相关性(P0.05)。结论ataxin-3可能参与卵巢低级别浆液性囊腺癌的发生机制,可能成为其早期诊断指标和治疗靶点。     

Nuclear findings of ovarian surface epithelial tumors

While cytoplasmic features of ovarian surface epithelial tumors are well-known, the nuclear findings have received little attention. We reviewed imprint cytology materials of the ovary which were collected at the Kawasaki Medical School Hospital between January 1989-July 1999, and identified 15 mucinous cystadenomas, 3 borderline mucinous tumors, 4 mucinous cystadenocarcinomas, 4 serous cystadenomas, 4 borderline serous tumors, 7 serous cystadenocarcinomas, 6 endometrioid carcinomas, and 2 clear-cell adenocarcinomas. We microscopically observed nuclear findings of the 45 cases. Coffee-bean nuclei were observed in 15.0%, 15.8%, and 10.1% of the tumor cells in mucinous adenomas, borderline mucinous tumors, and borderline serous tumors, respectively. The frequencies of the coffee-bean nuclei in the three tumors were higher than in the remaining tumors (P < 0.001). Intranuclear cytoplasmic inclusions were observed in 2.1% of the tumor cells in mucinous cystadenoma, and their frequency was significantly higher than that in cases of other surface epithelial ovarian tumors (P < 0.001). Semilunar-shaped nuclei were seen in all cases of mucinous cystadenomas and borderline mucinous tumors, and in 3 of 4 mucinous adenocarcinomas. The remaining surface epithelial tumors did not reveal the semilunar-shaped nuclei. In the cytology of the ovary, the semilunar nuclei are characteristic of mucinous tumors, and the intranuclear cytoplasmic inclusion may be a diagnostic clue to mucinous cystadenoma, when it is conspicuous. The coffee-bean nuclei can be seen in mucinous cystadenoma, borderline mucinous tumors, and borderline serous tumors.     

Aberrant hypermethylation of RASSF1A promoter in ovarian borderline tumors and carcinomas

The newly identified 3p21.3 tumor suppressor gene RASSF1A is inactivated by hypermethylation in variable solid tumors, including those of the lung, breast, prostate, kidney, and ovary. The purpose of this study was to evaluate the methylation status of RASSF1A in various types and stages of ovarian epithelial tumors. We analyzed the DNA methylation status of ovarian tumors using methylation-specific polymerase chain reaction in 54 frozen ovarian tumor tissues and in 97 cases of archival ovarian serous epithelial tumors using a microdissection procedure. Hypermethylation statuses were examined vs clinicopathologic findings. RASSF1A promoter methylation rates in the various types of fresh ovarian tissues were as follows: serous cystadenoma (1/5), serous tumor of borderline malignancy (2/7), serous adenocarcinoma (4/10), mucinous cystadenoma (0/5), mucinous tumor of borderline malignancy (2/7), mucinous adenocarcinoma (3/6), transitional-cell carcinoma (1/3), clear-cell carcinoma (3/3), and malignant müllerian mixed tumor (3/3). In archived serous tumor tissues, RASSF1A promoter hypermethylation was detected in serous cystadenoma (1/6, 16.6%), serous tumor of borderline malignancy (20/41, 48.8%), and in serous adenocarcinoma (25/50, 50%). The status of RASSF1A hypermethylation in borderline tumors was found to correlate statistically with the presence of microinvasion (p=0.002), peritoneal implant (p<0.001), and bilaterality (p=0.019). The RASSF1A promoter hypermethylation was frequently found in borderline tumors and carcinomas, suggesting that RASSF1A promoter hypermethylation may be a useful molecular marker for the early detection of ovarian tumors.     

Amylase in fallopian tube and serous ovarian neoplasms: immunohistochemical localization

We studied the cellular localization of amylase in normal Fallopian tubes and serous ovarian neoplasms using an indirect immunoperoxidase technique. The primary antiserum was against human pancreatic amylase, and was found to inhibit ovarian tumor amylase as well. Amylase was present in normal endosalpingeal epithelium and in the epithelial cells of benign, borderline, and malignant serous ovarian tumors. A mucinous cystadenoma was also studied and contained no amylase. This localization suggests that amylase production by serous ovarian neoplasms reflects the endosalpingeal differentiation of these tumors. Antibody to amylase may be potentially useful in distinguishing serous ovarian tumors from other forms of ovarian neoplasia.     

Mucinous borderline ovarian tumors: points of general agreement and persistent controversies regarding nomenclature, diagnostic criteria, and behavior

This report focuses on the borderline category of ovarian mucinous tumors and summarizes the points of general agreement and persistent controversies identified by experts in the field who participated in the Borderline Ovarian Tumor Workshop held in Bethesda, MD, in August 2003. Points of agreement and persistent controversies regarding nomenclature, diagnostic criteria, and behavior are addressed for the following ovarian mucinous tumor categories: mucinous borderline ovarian tumor (M-BOT; synonymously referred to as atypical proliferative mucinous tumor of ovary or mucinous ovarian tumor of low malignant potential), M-BOT with intraepithelial carcinoma, and M-BOT with microinvasion. The morphologic spectrum of M-BOTs with regard to distinction from mucinous cystadenoma and the confluent glandular/expansile type of invasive mucinous carcinoma is also addressed. Non-ovarian mucinous tumors, including the secondary ovarian mucinous tumors associated with pseudomyxoma peritonei and metastatic mucinous carcinomas with a deceptive pattern of invasion, are recognized as tumors that can simulate primary M-BOTs. Improved classification of these mucinous tumors has clarifed the behavior of true M-BOTs by excluding these simulators from the M-BOT category.     

Expression and mutational analysis of c-kit in ovarian surface epithelial tumors

Coexpression of Kit ligand and c-kit has been reported in some gynecologic tumors. To determine whether imatinib mesylate is useful in ovarian epithelial tumors, we performed immunohistochemical and mutational analysis. The cases consisted of 33 cases, which included 13 serous cystadenocarcinomas, 1 borderline serous tumor, 8 mucinous cystadenocarcinomas, 6 borderline mucinous tumors and 5 clear cell carcinomas. Five cases of serous cystadenoma and 5 cases of mucinous cystadenoma were also included. In the immunohistochemical study, 3 cases (3/6, 50%) of borderline mucinous cystic tumor and two cases (2/8, 25%) of mucinous cystadenocarcinoma show positive staining for KIT protein. Only one case (1/13, 7.7%) of serous cystadenocarcinoma had positive staining. On mutational analysis, no mutation was identified at exon 11. However, two cases of borderline mucinous tumors and one case of mucinous cystadenocarcinoma had mutations at exon 17. In these cases, the immunohistochemistry also shows focal positive staining at epithelial component. Although, KIT protein expression showed higher incidence in mucinous tumors than serous tumors, they lack KIT-activating mutations in exon 11. Thus, ovarian surface epithelial tumors are unlikely to respond to imatinib mesylate.     

卵巢粘液性囊腺瘤恶变和肠上皮化生的关系

Ninety cases of ovarian mucinous tumors were studied histologically and histochemically. Intestinal metaplasia was found in 48.2% (14/29) of benign. 73.6% (14/19) of borderline and 92.9% (39/42) of malignant mucinous cystadenomas. The differences between these three groups are statistically significant (P less than 0.01). Among 67 cases of intestinal metaplastic mucinous tumors, 43 contained argyrophil cells, and 36 contained argentaffin cells. The coexistence of intestinal metaplasic and uterocervical canal type epithelia was observed in 2/3 of borderline and 1/3 of malignant intestinal mucinous cystadenomas. In addition, there were 5 cases of borderline and 3 cases of malignant uterocervical canal type mucinous cystadenomas among the 90 cases. It is evident that the malignant transformation of ovarian mucinous cystadenoma was closely related to intestinal metaplasia. Anyhow, it seems not necessary for malignant transformation of all ovarian mucinous cystadenomas to pass through a stage of intestinal metaplasia: some of the malignant mucinous cystadenomas were considered probably originating from the uterocervical canal type epithelium.     

Mucin gene transcripts in benign and borderline mucinous tumours of the ovary: an in situ hybridization study

Mucinous tumours of the ovary are characterized by mucin-secreting cells exhibiting a variable endocervical, intestinal, gastric or pancreatobiliary phenotype as ascertained by microscopy, electron microscopy, histochemistry or immunohistochemistry. The molecular mechanisms underlying the tumourigenesis process are not well understood. The mucin glycoproteins expressed by ovarian mucinous tumours have not been fully characterized, but mucins are known to be implicated in tumour progression in various epithelial neoplasms. The purpose of this study was to evaluate the expression of mucin genes (MUC1, MUC2, MUC3, MUC4, MUC5AC, MUC5B, MUC6) in ovarian mucinous tumour cells, to relate MUC gene expression to the histological diagnosis, and to compare the expression patterns with those observed in normal tissues. The expression of mucin genes was evaluated by in situ hybridization in 21 mucinous tumours (11 adenomas and ten borderline tumours). Heterogeneity of expression correlated with morphological heterogeneity. Intense expression of the MUC5AC gene, suggesting a gastric surface cell phenotype, was demonstrated in 18/21 tumours (86%). Goblet cells expressing the MUC2 gene and columnar cells expressing the MUC3 gene were consistent with an intestinal phenotype, which was observed in 15 tumours (71%) including nine adenomas and six borderline tumours. Major expression of MUC4 and MUC5B consistent with an endocervical phenotype was observed in seven benign (64%) and three borderline (30%) tumours. In all, the MUC profiles suggested gastrointestinal-type cells in 13 cases (62%), gastric-type cells in five cases (24%), and intestinal-type cells in two cases (one benign, one borderline) (9%); the results were inconclusive in one borderline tumour (5%). It is concluded that gastric and, to a lesser degree, intestinal differentiation are early and almost constant events in ovarian mucinous tumourigenesis.     

Metastatic neoplasms involving the ovary: a review with an emphasis on morphological and immunohistochemical features

The ovary is a common site of metastatic tumour. In many cases of ovarian metastasis there is a known history of malignancy but in other cases the ovarian tumour is the first manifestation of disease. In this review metastatic colorectal, appendiceal, gastric, breast, pancreatic and biliary tract, hepatocellular, renal, transitional and cervical carcinomas and metastatic malignant melanoma involving the ovary are discussed, as is the issue of synchronous ovarian and endometrial carcinomas. Peritoneal tumours, including primary peritoneal carcinoma, mesothelioma and intra-abdominal desmoplastic small round cell tumour, involving the ovary are also discussed, together with a variety of other rare, metastatic ovarian neoplasms. Many metastatic adenocarcinomas involving the ovary, especially those exhibiting mucinous differentiation, closely mimic primary ovarian adenocarcinomas with morphologically bland areas simulating benign and borderline cystadenoma. This is referred to as a maturation phenomenon. In recent years immunohistochemistry, especially but not exclusively differential cytokeratin (CK7 and CK20) staining, has been widely used as an aid to distinguish between a primary and secondary ovarian adenocarcinoma. While immunohistochemistry undoubtedly has a valuable role to play and is paramount in diagnosis in some cases, the results must be interepreted with caution, especially in mucinous tumours, and within the relevant clinical context. We feel the significance of differential cytokeratin staining is not always understood by histopathologists and this can result in erroneous interpretation. We critically discuss the value of immunohistochemistry and associated pitfalls with each tumour type described.