Morphology of serous borderline ovarian tumors

The paper provides the results of the examination of ovarian serous borderline tumors in 68 patients followed up during different periods (up to 22 years) after surgery. More valuable information on the course and prognosis of the disease will be provided by taking into account both the process stages and structural features of extra-ovarian implants rather than by detecting the foci of initial invasive growth in the primary tumor. Among 4 peritoneal neoplasia types defined by the author, the desmoplastic reaction occurring around the tubal type tumor cells in particular, should be regarded as the most unfavourable type. The author believes that the serous borderline tumor of the ovary is not a separate nosological entity but a step in progressive malignant transformation of mesothelial tubal epithelium similar to definition of well differentiated carcinoma given by the traditional classification.     

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。     

Genomic imbalances in ovarian borderline serous and mucinous tumors

We analyzed 25 ovarian borderline tumors (13 serous and 12 mucinous tumors) by comparative genomic hybridization (CGH). Genomic imbalance was detected in 85% of serous tumors and 75% of mucinous tumors. Different patterns of genomic alterations were identified in serous and mucinous tumors. Gain of the X chromosome was common in both serous (30%) and mucinous (42%) tumors. However, gain of chromosome 8 was detected exclusively in 38% of serous and mixed sero-mucinous tumors, but not in any pure mucinous tumors. According to the present and previous studies, gain of chromosome 8 is the most common abnormality in borderline serous tumors. Gain of the same chromosome is also common in high grade and advanced stage serous carcinomas, but uncommon in early stage serous carcinomas. In addition gain of chromosome X is common in borderline serous and mucinous tumors, while loss of chromosome X is predominant in invasive carcinomas. These findings do not support the multi-step progression theory from borderline tumor to high-grade, advanced stage carcinoma, but indicate that the borderline ovarian tumor is a distinct entity. Genes in chromosome 8 may be critical for the development and the differentiation of borderline serous tumors.     

卵巢浆液性和粘液性交界瘤的临床病理分析

目的：观察卵巢交界瘤的临床病理学特点,探索肿瘤不同组织学改变的意义。方法：对45例卵巢浆液性和粘液性交界瘤进行回顾性分析,肿瘤分期按国际妇产科联合会（FIGO）标准,Ⅰ期34例,Ⅱ期4例,Ⅲ期7例。结果：45例卵巢浆液性和粘液性交界瘤占同期卵巢上皮恶性肿瘤的25．4％,浆液性同粘液性交界瘤的比为1：1．3,11例生长于卵巢表面的浆液性交界瘤中,9例出现腹膜种植,2例为浸润性种植,7例为非浸润性种植。2例浆液性交界瘤和1例粘液性交界瘤分别于术后5、4和1年复发。33例交界瘤经2－9年随访,按Kaplan-Meier法5年生存率为100％。结论：卵巢浆液性交界瘤预后较好,卵巢表面生长的浆液性交界瘤常伴有腹膜种植。     

Differential diagnosis of borderline serous and mucinous ovarian tumors

Electron microscopic study of borderline serous and mucinous ovarian tumours was carried out and revealed marked heterogeneity of both types of these tumours. On the basis of the remote results of surgical treatment of the patients the most dangerous subtype from the point of view of its malignancy, the so-called proliferating, tumour with atypical epithelium, was established.     

卵巢浆液性交界及恶性肿瘤K-ras基因突变分析

目的探讨K-ras基因在卵巢浆液性交界及恶性肿瘤发生发展过程中的作用。方法收集51例卵巢浆液性肿瘤标本,包括经典型交界性浆液性肿瘤18例,微乳头型交界性浆液性肿瘤11例,浸润性微乳头型浆液性癌12例,经典型浆液性癌10例。采用显微切割技术获取肿瘤细胞后,提取基因组DNA、PCR技术扩增K—ras基因第一外显子,通过直接测序的方法鉴定K—ras基因第12、13密码子的突变情况。结果1例经典型交界性浆液性肿瘤K—ras基因第12密码子发生突变,突变类型为GGT→GTT即甘氨酸→缬氨酸,余50例标本未见突变;所有标本K—ras基因第13密码子均为野生型。结论K—ras基因第12、13密码子在被检患者中卵巢浆液性交界及恶性肿瘤中的突变频率很低,其在该肿瘤发生发展过程中可能不起主要作用。     

Molecular genetic evidence for monoclonal origin of bilateral ovarian serous borderline tumors

Patients with serous borderline tumors of the ovary often present with multiple tumors at different sites in the abdominal cavity. Whether different foci of ovarian serous borderline tumors are monoclonal in origin, arising as a consequence of spread from a single ovarian site, or whether such deposits are polyclonal and explained by independent molecular genetic alterations on the background of a field defect, is unknown. So far, only X-chromosome inactivation studies were performed to study this issue. We used a genome-wide allelotyping to assess clonality in 47 metachronous and/or synchronous multifocal tumors from 22 patients, using 59 microsatellite markers. Loss of heterozygosity (LOH) was observed in only 34 of 1969 informative markers in 9 of 22 serous borderline cases studied. Of these cases, 7 showed concordant LOH for at least one polymorphic marker in more than one tumor site. Flanking microsatellite markers enabled identification of identical chromosomal breakpoints in 6 of 7 cases. The LOH results strongly favor a common origin indicated by a likelihood ratio (possibility common origin/possibility independent origin) ranging from 39 to 14,163. Strong additional evidence for monoclonality is provided by the finding of identical microsatellite alterations in all three-tumor sites in one case.     

卵巢浆液性及黏液性交界性肿瘤的研究进展

长期以来对卵巢交界性肿瘤（borderline ovarian tumors,BOT）命名及病理诊断标准一直存在不同的意见。经过30余年的探索,对其本质认识逐步深入,特别是2003年8月在美国马里兰州贝塞斯达举行的卵巢交界性肿瘤工作会议,就卵巢交界性肿瘤的某些病理学方面的分歧取得了较一致的意见。本文就近年卵巢浆液性及黏液性交界性肿瘤的命名、形态学特征、病理诊断以及预后等方面的进展作如下综述。     

K-RAS mutations in ovarian and extraovarian lesions of serous tumors of borderline malignancy

K-RAS mutations are the most frequent molecular genetic alteration in serous ovarian tumors of borderline malignancy (SBOT). The pathogenesis of associated contralateral tumors and extraovarian implants and Müllerian inclusion cysts is obscure. We hypothesized that the comparison of K-RAS mutations in these lesions might help to distinguish multifocal from metastatic foci. Eight cases of SBOT with known K-RAS mutation (RAS+) and two cases without mutation (RAS-) were analyzed for comparison. DNA was extracted from multiple samples of 58 paraffin-embedded and laser-microdissected ovarian and extraovarian lesions (10 ovarian borderline tumors, 8 contralateral tumors, 25 implants, 15 inclusion cysts; total: 97 samples). K-RAS exon 1 was amplified by PCR and analyzed by denaturing gradient gel electrophoresis and cycle sequencing. In 12 of 14 SBOT and in 2 of 2 extraovarian implants, the K-RAS mutation could be found in different areas of the same lesion, indicating monoclonality. All RAS+ ovarian borderline tumors with contralateral tumors (six of six) harbored an identical mutation in both ovaries (in one case, a separate surface borderline tumor component contained a different mutation in addition). In 4 of 5 RAS+ ovarian tumors with extraovarian lesions, RAS mutations were also found in implants (15/21 implants [71%]) and more rarely in inclusion cysts (3 of 12 lesions [25%]). These extraovarian mutations were always identical to the one in the ovary (18 of 18 [100%]). Regarding the contralateral and extraovarian lesions of the two RAS- SBOT, only one extraovarian implant contained a RAS mutation. The demonstration of K-RAS mutations in Müllerian inclusion cysts and implants of SBOT suggests that K-RAS mutations represent a pivotal event during neoplastic transformation of ovarian and extraovarian serous epithelium. Considering our observations, the two putative pathogenetic mechanisms for the development of implants and endosalpingiosis-multifocal tumorigenesis and spread from the ovarian primary tumor-seem to coexist.     

Genetic alterations of serous borderline tumors of the ovary compared to stage I serous ovarian carcinomas

Borderline tumors of the ovary comprise 10-20% of all epithelial ovarian tumors, and are placed between clearly benign and obviously malignant ovarian tumors. The issue of whether borderline tumors are precursors of invasive carcinoma or distinct clinical entities, however, is still the subject of discussion. To increase our understanding in relation to this issue, the aim of our study was to analyze both serous borderline and invasive ovarian tumors, and to investigate early carcinogenesis in serous ovarian tumors. Using comparative genomic hybridization, we compared cytogenetic changes in borderline ovarian tumors and stage I invasive tumors. The average number of genetic alterations differed significantly between the borderline and the invasive tumors (1.9 and 9.2, respectively). The most common genetic alterations among the borderline tumors were loss of chromosome 17, 20q, and 18p, and gain of 12p13 approximately q23. These changes were also found among the invasive tumors in a similar percentage. In conclusion, we found four distinct cytogenetic alterations that might be early events in serous ovarian tumors, and that might also characterize a subgroup of borderline ovarian tumors that may have the potential to progress and develop malignancy.     

High-grade ovarian serous carcinoma exhibits significantly higher p16 expression than low-grade serous carcinoma and serous borderline tumour

AIMS: A dualistic pathway of ovarian serous carcinogenesis is now well established whereby high-grade serous carcinoma and low-grade serous carcinoma represent two distinct tumour types with a different underlying pathogenesis. The aim of this study was to compare expression of p16 INK4A (p16) in these two tumour types. We also included cases of serous borderline tumour, since these are considered to represent a precursor lesion of low-grade serous carcinoma. METHODS AND RESULTS: Cases of serous borderline tumour (n = 18), low-grade ovarian serous carcinoma (n = 22) and high-grade ovarian serous carcinoma (n = 24) were stained with a monoclonal antibody against p16. Cases were scored both with respect to intensity of immunoreactivity (weak, 1+; moderate, 2+; or strong, 3+) and distribution (0, negative or occasional positive cells; 1+, < 10% cells positive; 2+, 10-25% cells positive; 3+, 26-50% cells positive; 4+, 51-75% cells positive; or 5+, 76-100% cells positive). An immunohistochemical composite score was also calculated (0-15) by multiplying the intensity and distribution scores. There was a statistically significant difference in p16 immunoreactivity with respect to intensity, distribution and composite score between high-grade serous carcinoma and each of the other two groups, with the high-grade neoplasms exhibiting stronger and more diffuse positivity. Most high-grade serous carcinomas exhibited positivity of close to 100% of tumour cells. There was no significant difference in p16 expression between the borderline tumours and low-grade serous carcinomas. CONCLUSIONS: The increased expression of p16 in high-grade serous carcinoma compared with low-grade serous carcinoma and serous borderline tumour is in keeping with a different underlying pathogenesis. p16 may be implicated in the development of high-grade serous neoplasia within the ovary and elsewhere within the female genital tract.     

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。     

Metallothionein expression and nuclear size in benign,borderline, and malignant serous ovarian tumours

Metallothioneins (MTs) are low-molecular-weight proteins involved in metalloregulatory functions such as cell proliferation, growth, and differentiation. In recent years, MT expression has been linked with carcinogenesis, resistance to cancer therapy, and tumour progression. However, the significance of MT expression in ovarian cancers is at present inadequately documented. In this study, MT immunohistochemistry was performed in 12 benign, 14 borderline, and eight malignant serous tumours of the ovary. The intensity of the immunostaining was evaluated by image analysis. There was a significantly higher number of MT-immunopositive cells in the multilayered epithelial cells of borderline serous tumours (atypical proliferative serous tumours) than in the single layered epithelial cells within the same tumour, and in the single cell layer of benign serous tumours. There was no difference in the expression of MTs in the single layered tumour cells of benign and borderline serous tumours. Significantly higher numbers of MT-immunopositive cells were observed in both the single and the multilayered epithelial cells of serous carcinomas, the highest number being observed in the multiple layers of serous carcinomas. The positively stained malignant tumour cells in both single and multiple layers were larger than the negatively stained cells in benign, borderline, and malignant serous ovarian tumours. There was moderate to intense staining. These findings indicate that there is increased expression of MTs in the progression of malignancy, which could be used as a marker in grading the three groups of ovarian serous tumours and for determining prognosis. Copyright © 1999 John Wiley & Sons, Ltd.     

Overexpression of trophoblast cell surface antigen 2 as an independent marker for a poor prognosis and as a potential therapeutic target in epithelial ovarian carcinoma

Most patients with epithelial ovarian cancer (EOC) are diagnosed at an advanced stage, and therapeutic options for these patients are limited. The identification of suitable biomarkers could be helpful for patients with EOC, who might benefit from targeted therapies even in advanced stages of the disease. Trophoblast cell surface antigen 2 (TROP2) is highly expressed in various human malignant tumours; however, this has not been demonstrated clearly in EOC. In this study, we further evaluated whether TROP2 is a promising marker for EOC, and thus also a potential target for EOC immunotherapy. Quantitative real‐time polymerase chain reaction (qPCR) and fluorescence‐activated cell sorting (FACS) analysis were employed to determine TROP2 mRNA and protein expression in both human EOC and normal ovarian cell lines. Additionally, TROP2 protein expression was measured by immunohistochemistry in 128 EOC tissue samples, 21 normal ovarian tissues and 18 normal fallopian tubes. The correlations between TROP2 protein expression and patients' clinicopathological features were investigated, and survival outcomes were analysed. TROP2 mRNA and protein levels were upregulated significantly in EOC cell lines compared with normal cell lines. The protein of TROP2 was expressed in the majority of EOC tissue samples (90.6%) and overexpressed in 75 (58.6%) of the 128 tumour samples. TROP2 overexpression was correlated with relevant clinicopathological characteristics and was associated with significantly shortened overall survival and disease‐free survival. Furthermore, TROP2 was an independent prognostic marker for EOCs as analysed by Cox regression. TROP2 was a potential biomarker for targeted therapy in patients with TROP2‐overexpressing EOC.     

Overexpression of p21WAF1/CIP1 and MDM2 characterizes serous borderline ovarian tumors

Ovarian epithelial tumors are classically divided into benign, malignant, and borderline or of low malignant potential. It is controversial whether this last group of tumors should be considered benign or malignant. Expression of cell cycle markers has recently been linked to tumor behavior and response to treatment. It has been shown that one of the pathways through which the p53 gene controls the cell cycle is by transactivating p21WAF1/CIP1, a cyclin-dependent kinase (cdk) inhibitor. By inhibiting cdks, p21WAF1/CIP1 blocks the G-1 to S-phase transition in the cell cycle. p53 can be regulated by MDM2 (murine double minute-2) through direct inactivation or promotion of its cytoplasmic degradation. In an attempt to investigate the cell cycle checkpoint mechanisms of these tumors, we studied the expression of p53, Ki-67, MDM2, and p21WAF1/CIP1 by immunohistochemistry. We analyzed the expression of these proteins in 19 cystadenomas (8 serous and 11 mucinous), 40 borderline tumors (31 serous and 9 mucinous), and 18 serous carcinomas of the ovary. p21WAF1/CIP1 was expressed in 7 of 19 (37%) benign cystadenomas, 32 of 40 (80%) borderline tumors (93.5% of serous and 33% of mucinous), and in 9 of 18 (50%) serous carcinomas. Ki-67 was only weakly expressed in 8 of 19 (42%) benign cystadenomas, all borderline tumors showed Ki-67 staining in less than 50% of the cells, and 55% of serous carcinomas stained in more than 50% of tumor cells. p53 was absent in all but 1 of the cystadenomas, was expressed in 9 of 40 (22.5%) borderline tumors (25.8% of serous and 11% of mucinous), and in 10 of 18 (55%) carcinomas. All 11 implants of serous borderline tumors expressed p21WAF1/CIP1. Most serous borderline tumors expressed higher levels of MDM2 compared with the benign cystadenomas and carcinomas. Four of the serous borderline implants (40%) expressed MDM2. Coexpression of p21WAF1/CIP1 and MDM2 characterizes serous borderline tumors of the ovary and their implants, which suggests that these cell cycle control proteins are important in these tumors and may be related to tumor progression. Low expression of p53 protein in serous borderline tumors might be in part mediated by MDM2. This suggests that the p53 pathway is intact in most of these tumors, in contrast with carcinomas, in which high expression of p53 has been related to mutations of this gene.