Alterations in DNA methylation are early, but not initial, events in ovarian tumorigenesis.

We compared global levels of DNA methylation as well as methylation of a specific locus (MyoD1) in ovarian cystadenomas, ovarian tumours of low malignant potential (LMP) and ovarian carcinomas to investigate the association between changes in DNA methylation and ovarian tumour development. As we realized that cystadenomas showed different methylation patterns from both LMP tumours and carcinomas, we verified their monoclonal origin as a means of confirming their true neoplastic nature. High-pressure liquid chromatographic (HPLC) analyses showed that global methylation levels in LMP tumours and carcinomas were 21% and 25% lower than in cystadenomas respectively (P = 0.0001 by one-way variance analysis). Changes in the methylation status of the MyoD1 locus were not seen in any of ten cystadenomas analysed but were present in five of ten LMP tumours and in five of ten carcinomas (P = 0.03). These findings suggest that alterations in DNA methylation are absent (or at least not as extensive) in ovarian cystadenomas, but are present in LMP tumours, the phenotypic features of which are intermediate between those of benign and malignant ovarian tumours. The results also emphasize the merit of distinguishing ovarian LMP tumours from cystadenomas, in spite of their similar clinical characteristics.

WAF1/CIP1 structural abnormalities do not contribute to cell cycle deregulation in ovarian cancer.

Mutations in the WAF1/CIP1 gene were not found in 36 ovarian carcinomas, including tumours with loss of heterozygosity at the WAF1/CIP1 locus and/or lacking p53 mutations. In addition, no association was demonstrable between a polymorphism in a conserved region of the WAF1/CIP1 gene and ovarian carcinoma.

Potential role of the inactivated X chromosome in ovarian epithelial tumor development.

BACKGROUND: Ovarian epithelial tumors can be divided into subcategories often regarded as different stages of neoplastic transformation. Cystadenomas belong to the least aggressive subgroup and are noninvasive and nonmetastatic. Ovarian tumors of low malignant potential (LMP) are intermediate between cystadenomas and carcinomas and show markedly reduced invasive and metastatic abilities. Invasion and metastasis are the hallmarks of carcinomas, which constitute the most aggressive subgroup and can be further subdivided into different grades. PURPOSE: We performed comparative allelotype analyses of ovarian cystadenomas, LMP tumors, and carcinomas, reasoning that such analyses could provide clues about the molecular determinants of their phenotypic differences. Because we realized that allelic losses involving the X chromosome might be associated with LMP tumor development, we determined whether such losses were interstitial and whether they involved the active or the inactive X chromosome. METHODS: Frequencies of loss of heterozygosity (LOH) at specific loci in every chromosomal arm were determined in 16 ovarian cystadenomas, 23 ovarian LMP tumors, 15 low-grade ovarian carcinomas, and 35 high-grade ovarian carcinomas by use of either the polymerase chain reaction (PCR) or Southern blot analyses. We took advantage of the fact that DNA methylation is an important mechanism of X-chromosome inactivation to determine whether losses involving the X chromosome were in the active or the inactive copy. We analyzed the methylation status of retained alleles on the X chromosome by determining whether they could be amplified by PCR after digestion with the methylation-sensitive restriction endonuclease Hpa II. RESULTS: High-grade carcinomas contained frequent(>50%) LOH in four autosomal chromosome arms, i.e., 6q, 13q, 17p, and 17q. Except for 13q, these same chromosomal arms showed frequent LOH in low-grade carcinomas. LOH in autosomal chromosomes was comparatively rare in LMP tumors and was absent in cystadenomas. In contrast, half (eight of 16) of LMP tumors informative for a locus in the proximal portion of chromosome Xq showed LOH at that locus. These losses were the result of interstitial deletions in six of the eight cases and involved the inactive copy of the X chromosome exclusively. Similar losses in the X chromosome were not seen in either cystadenomas or low-grade carcinomas. CONCLUSIONS AND IMPLICATIONS: LOH at multiple loci is associated with the development of ovarian carcinomas but not with the development of cystadenomas and LMP tumors. However, the integrity of a locus in chromosome Xq that possibly escapes X-chromosome inactivation is important for the control of LMP tumor development. The fact that this locus does not appear to be involved in the genesis of low-grade carcinomas suggests that LMP tumors are not precursors of such carcinomas.

Genetic disparity between morphologically benign cysts contiguous to ovarian carcinomas and solitary cystadenomas.

BACKGROUND: Ovarian carcinomas occasionally contain large, histologically benign cysts contiguous to the clearly malignant areas (cystadenocarcinomas). The question of whether such cysts are remnants of pre-existing benign tumors (cystadenomas) or constitute integral components of the carcinomas is important in clarifying the role of cystadenomas in ovarian carcinogenesis. It is also important for our general understanding of tumor heterogeneity, a phenomenon thought to result from the gradual accumulation of genetic abnormalities in initially homogeneous tumors. This question is also pertinent to the clinical management of ovarian cystadenomas, which are frequent in women of childbearing age and are usually treated surgically based on the possibility that they may give rise to carcinomas. PURPOSE: Reasoning that molecular markers of ovarian malignancy would be confined to the histologically malignant portions of cystadenocarcinomas if the morphologically benign portions are in fact pre-existing typical cystadenomas, we sought to verify that mutations in the p53 tumor suppressor gene are markers of malignancy in ovarian tumors and to determine the distribution of such mutations in cystadenocarcinomas. METHODS: We used immunohistochemical and DNA-sequencing techniques to analyze 46 ovarian carcinomas, 21 ovarian tumors of low malignant potential, and 16 solitary cystadenomas for the presence of p53 mutations. We then used similar techniques to examine the distribution of such mutations in different portions of cystadenocarcinomas. The observed differences in mutation frequencies were analyzed by the two-tailed Fisher's exact test. RESULTS: Mutations in the p53 gene were present in 24 (52%) of the 46 carcinomas, but they were absent in the 21 tumors of low malignant potential (P < .0001) and the 16 solitary cystadenomas (P = .0002). Six of six cystadenocarcinomas with p53 mutations showed the presence of the same mutations in the adjacent, histologically benign cysts. The mutations were seen not only in cells immediately adjacent to the carcinomas, but also throughout the morphologically benign cysts. Twenty (83%) of the 24 cases showing mutation of one p53 allele also showed loss of genetic heterozygosity, suggesting that the other p53 allele was deleted. Such allelic loss, if present in morphologically malignant portions of cystadenocarcinomas, was also observed in the contiguous cysts. CONCLUSIONS: Ovarian carcinomas can be distinguished from ovarian cystadenomas and tumors of low malignant potential by p53 mutations. The fact that the mutations were present in histologically benign cysts contiguous to ovarian carcinomas suggests that such cysts are not typical cystadenomas and may carry a genetic predisposition to carcinogenesis that is not present in ordinary cystadenomas.

Inhibition of adhesion reformation in the rabbit model by meclofenamate: an inhibitor of both prostaglandin and leukotriene production.

OBJECTIVE: To determine the relative ability of meclofenamate sodium, a water-soluble inhibitor of both prostaglandin and leukotriene synthesis, to inhibit adhesion reformation. DESIGN: Prospective, randomized study in a rabbit model. INTERVENTIONS: Laparotomies were performed on mature New Zealand White rabbits, and each uterine horn was devascularized and traumatized with unipolar electrocautery. One week later, adhesions were microsurgically lysed. Each rabbit was randomly assigned to one of five different groups, and different solutions or an adhesion barrier were placed into the peritoneal cavities before closure: [1] control, 40 mL of normal saline (n = 8); [2] meclofenamate, 1.75 mg/mL in 40 mL of normal saline (n = 7); [3] Hyskon, 40 mL of 32% dextran-70 (n = 6); [4] meclofenamate 1.75 mg/mL in 40 mL of 32% dextran-70 (n = 6); and [5] TC-7, 40 mL of normal saline plus oxidized regenerated cellulose fabric, Interceed, placed over the site of adhesion lysis (n = 6). Two weeks later, adhesion reformation was scored according to percent involvement of each uterine horn (0 to 4), and adhesion density (0 to 1) and compared using a one-factor analysis of variance. RESULTS: Adhesion reformation was greatest in the control group (mean score +/- SEM, 3.7 +/- 0.4) and was decreased, but not significantly, in the Hyskon group (2.7 +/- 0.4). Compared with the control group, reformation was significantly decreased in the meclofenamate group (2.3 +/- 0.2), the TC-7 group (2.0 +/- 0.5), and the meclofenamate/Hyskon group (1.1 +/- 0.3). This last group was also decreased compared with the meclofenamate and Hyskon groups. CONCLUSION: Meclofenamate significantly inhibits adhesion reformation in the rabbit model, especially when used in combination with a 32% dextran-70 solution.

Etiology of closure-related adhesion formation after wedge resection of the rabbit ovary.

Adhesion formation after closure after ovarian wedge resection of the rabbit ovary may be related to the use of polyglactin suture rather than to the closure itself.