Myometrial invasion and cervical invasion by endometrial carcinoma: evaluation by CO2-volumetric interpolated breathhold examination (VIBE).

PURPOSE: To assess myometrial invasion and cervical invasion by endometrial carcinoma, using CO2-volumetric interpolated breathhold examination (VIBE) enabling more precise evaluation of depth of tumor invasion. MATERIALS AND METHODS: CO2-VIBE was performed in 21 cases of endometrial carcinoma (Stage Ia-IIb) prior to treatment. The images were interpreted by performing multiplanar reconstruction (MPR), and the findings obtained from the images (degree of myometrial invasion and presence or absence of cervical invasion) were assessed in comparison with the histopathological diagnosis. RESULTS: The sites of the endometrial carcinoma lesions were clearly visualized by the CO2-VIBE method. Evaluation of the degree of myometrial invasion enabled a high correct diagnosis rate of 90.5%, and evaluation for the presence of cervical invasion also allowed a high correct diagnosis rate of 90.5%. CONCLUSION: VIBE permits evaluation of any plane desired by means of thin slices, and it is a truly revolutionary method for preoperative evaluation of depth of invasion of endometrial carcinoma that enables highly accurate determination of the extent of lesion sites and degree of invasion before treatment.

Establishment of a HPV and p53-mutation-negative human cell line (CA) derived from a squamous carcinoma of the uterine cervix.

OBJECTIVE: Well-characterized human cancer cell lines are important research resources for studying cancer cell biology, as well as for developing new strategies against cancer cell growth and progression. We present a new cell line, CA, established from an invasive non-keratinizing squamous cell carcinoma of the uterine cervix in 36-year-old patient. METHODS: We measured the doubling time of CA cells. To investigate the tumorigenicity of CA, cells were inoculated subcutaneously into the back of nude mice. Several tumor markers were analyzed using culture media by EIA. PCR-based analyses were performed to examine the human papillomavirus (HPV) status and telomerase activity. CA was also screened for p53 mutation using the sequencing technique. RESULTS: The cells show rapid growth in culture with a doubling time of 14.3 h and high migration activity. Monolayer-cultured cells were polygonal, showing a pavement-like arrangement and a tendency to pile up without contact inhibition. Subcutaneous transplantation of the CA cells into nude mice formed solid tumors that were histologically diagnosed as squamous cell carcinoma, whereas no metastasis was observed. Cultured CA cells produced SCC, CEA, TPA, CA125 and SLX. Genetic and molecular analyses revealed high telomerase activity and the absence of HPV DNA. No p53 mutation was observed in this cell line. CONCLUSION: These properties suggest that CA is an aggressive cervical carcinoma cell line and may serve as a useful experimental model for studying HPV role in cervical carcinogenesis.

Rupture of pregnant noncommunicating rudimentary uterine horn with fetal salvage: a case report.

A rare case of noncommunicating rudimentary uterine horn pregnancy is described. The pregnancy proceeded to twenty-six gestational weeks when the rudimentary uterine horn ruptured as the patient had signs and symptoms of massive hemoperitoneum. An emergency exploratory laparotomy revealed incomplete rupture of the gravid rudimentary horn. A viable female infant with a birth weight of 633 g was delivered. The rudimentary horn had no direct communication to the uterine cavity of the unicornate right uterus. Immunohistochemical examination showed that the excised uterine horn was filled with placental tissue without an intervening layer of decidua basalis.

A novel phenoxazine derivative suppresses proliferation of human endometrial adenocarcinoma cell lines, inducing G2M accumulation and apoptosis.

We examined the effects of a novel phenoxazine, 2-amino-4,4alpha-dihydro-4alpha,7-dimethyl-3H-phenoxazine-3-one (Phx), which was produced by the reaction of 2-amino-5-methyl-phenol with bovine hemoglobin on the proliferation of human endometrial adenocarcinoma cell lines, EN and KLE cells, and on induction of apoptosis and G2M arrest in these cells. Phx inhibited proliferation of these cell lines in a dose- and time-dependent manner, i.e., the inhibition rate of proliferation of EN and KLE cells was 43% and 40%, respectively, in the presence of 50 micro M Phx, and 75% and nearly 100%, in the presence of 100 micro M Phx, after 2 days. When these endometrial adenocarcinoma cells were incubated with a medium containing 100 micro M Phx for 24 h, accumulation of EN and KLE cells in the S and G2M phase and that of apoptotic cells were demonstrated by flow cytometry. Apoptosis of these cells caused by Phx was unlikely to be associated with p53, Bax, and Bcl-2, because the levels of these proteins were not altered regardless of the presence or absence of Phx. The present results suggest that Phx demonstrates antitumor activity against human endometrial adenocarcinoma cell lines EN and KLE cells, by inducing both cell cycle accumulation at S and G2M and apoptosis associated with p53, Bcl-2 and Bax insensitive pathways.

Increased expression of IgE-dependent histamine-releasing factor in endometriotic implants.

A complex network of cytokines mediates the immunoregulatory responses leading to endometriosis. Recent intensive studies suggest that monocyte and T cell chemoattractants contribute to the inflammatory environment of endometriotic implants. The relationship between the inflammation present during endometriosis and the development of endometriotic implants in the peritoneal cavity remains unclear. On the other hand, the association between endometriosis and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD; dioxin) exposure has been discussed in recent years, and our previous results revealed that IgE-dependent histamine-releasing factor (HRF) is inducible by TCDD. The present study aimed to clarify the expression, localization, and function of HRF in endometriosis. Northern blot analysis demonstrated that HRF is overexpressed in endometriotic implants. RT-PCR with Southern blot analysis, however, showed that HRF overexpression was not always accompanied by CYP1A1 induction in endometriotic implants, suggesting that HRF is inducible in endometriosis without exposure to TCDD. HRF is also inducible by macrophage colony-stimulating factor (M-CSF). Immunohistochemistry showed CD68-positive macrophages in the stroma of endometriotic implants, adjacent to regions with prominent HRF accumulation. HRF-overexpressing cells exhibited high implantation efficiency in comparison to control cells when the cells were injected into the peritoneal cavities of nude mice. These results suggest that the accumulation of macrophages in endometriotic implants induces HRF; the overexpression of HRF accelerates the growth of endometriotic implants.

Establishment of a new human cell line (EN) with TP53 mutation derived from endometrial carcinoma.

We present a new cell line, EN, established from an invasive endometrioid adenocarcinoma of the uterine corpus in 50-year-old patient. The cells show rapid growth in culture with a doubling time of 24.4 hours and high migration activity. Monolayer-cultured cells were polygonal in shape and showed a tendency to pile up without contact inhibition. Subcutaneous transplantation of the EN cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma, whereas no metastasis was observed. Cultured EN cells produced tissue polypeptide antigen. Genetic and molecular analyses revealed high telomerase activity and estrogen receptor beta but not alpha expression. Using the polymerase chain reaction-single strand conformation polymorphism technique, we have screened EN cells for TP53 mutation in exons 5-8. A mobility shift was observed in this cell line in exon 8. A nucleotide insertion (CGT-->CAGT) was detected at codon 273, which resulted in a creation of a stop codon at codon 308. This cell line thus appears to represent the development of a more malignant clone with divergent receptor function and growth behavior, and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium.

Matrix metalloproteinase-26 is expressed in human endometrium but not in endometrial carcinoma.

BACKGROUND: The human matrix metalloproteinase (MMP)-26, also called matrilysin-2 or endometase, has been isolated as a matrilysin (MMP-7) homolog. Matrix metalloproteinase-26 was expressed in tissue samples from the placenta and endometrial tumors and its expression may be related to the development of endometrial carcinomas. METHODS: Total RNAs were isolated from 5 endometrial carcinoma cell lines, 36 normal endometrial tissue samples, 4 hyperplasia tissue samples, and from 24 endometrial carcinoma tissue samples. Reverse transcription-polymerase chain reation (RT-PCR) was performed to detect MMP-26 mRNA expression. To identify MMP-26 mRNA localization and protein expression, we performed in situ RT-PCR and immunohistochemistry, respectively. RESULTS: Reverse transcription-polymerase chain reaction analysis revealed that MMP-26 mRNA was expressed in 24 of 36 normal human endometrial tissue samples. However, MMP-26 mRNA expression was not detected in endometrial carcinoma cell lines nor in endometrial carcinoma tissue samples except for one case. Western blot analysis showed similar results. In situ RT-PCR analysis revealed that MMP-26 expression was localized in the epithelial glandular cells but faint expression was observed in the stromal cells. Subsequently, we separated endometrial tissues into epithelial glandular and stromal cells. Using RT-PCR, the purified epithelial glandular cells exhibited MMP-26 mRNA expression but the purified stromal cells did not. Immunohistochemical analyses revealed that MMP-26 protein expression is also limited to endometrial epithelial glandular cells but not to cancer cells. Therefore, MMP-26 expression is limited to normal epithelial glandular cells. CONCLUSIONS: We found a significant difference in MMP-26 expression in normal and malignant endometrial tissue samples, although its function is still unknown. These data suggest that MMP-26 may be a candidate for a new tumor marker for endometrial carcinomas.

Evaluation of endometrial cancer with 3D-VIBE (volume interpolated breath-hold examination) using intrauterine CO2 gas.

To evaluate the myometrial infiltration of the endometrial cancer prior to aggressive treatment, dynamic MRI (magnetic resonance imaging) has attracted attention. However, it has also been found that in a number of cases, MRI exhibits inconsistent results with regards to the extent of the infiltration into this component of the uterine body. To overcome this limitation, the authors designed a method to delineate the tumor morphology more clearly by injecting CO2 gas into the uterine cavity. This procedure was combined with VIBE (volumetric interpolated breath-hold examination) to determine more precisely the depth of the tumor invasion. From our clinical results, the efficacy of the method was evaluated. On four patients with endometrial cancers (stage Ia-Ic), CO2 was injected to dilate the intra-uterine space through a catheter equipped with a balloon that had been introduced into the uterine cavity, after which VIBE was conducted. The images were interpreted by MPR (multiplanar reconstruction) and the findings from these images were compared against the histopathological findings. By employing this method, it was possible to delineate clearly the tumorous lesion in the uterine body, and three-dimensional images of the tumor invasion was acquired. The site and extent of tumor invasion in the myometrium were generally consistent with the histopathological findings. This method allows one to observe multiple planes by using thin slices. By dilating the uterine cavity, the site of involvement and the extent of invasion can be more precisely defined before treatment. It is truly a revolutionary procedure for determining-prior to surgery-the depth of invasion of a cancer located in the uterine body.

Differentially regulated genes as putative targets of amplifications at 20q in ovarian cancers.

Frequent amplification of DNA at 20q or part of 20q has been demonstrated by comparative genomic hybridization in ovarian cancer (OC), but the genetic target(s) of these amplification events remain unknown. We examined copy-number changes with respect to six candidate genes, E2F1 (20q11.2), TGIF2 (20q11.2), AIB1 (20q12), PTPN1 (20q13.1), ZNF217 (20q13.2), and BTAK (20q13), and then measured transcription levels of each candidate in 18 OC cell lines. Three distinct cores of amplification were identified: 20q11.2, harboring E2F1 and TGIF2 (region I; 1 of 18 cell lines, 5.6%); 20q13.1, harboring PTPN1 (region II; 5 lines, 27.8%); and 20q13.2, harboring ZNF217 and BTAK (region III; 6 lines, 33.3%). Among the six genes examined, expression levels of PTPN1 and ZNF217 were significantly correlated with absolute copy-number, and those of PTPN1 and TGIF2 were significantly correlated with copy-number relative to the centromere of chromosome 20 (20cen). Among 19 primary OCs examined, moreover, we observed amplification of TGIF2, PTPN1 and ZNF217 in five (26.3%), ten (52.6%), and twelve (63.2%) tumors, respectively. Expression levels of PTPN1 and ZNF217 were significantly correlated with their copy-numbers in those primary OCs. Our results suggest that 20q amplifications in OCs can be extensive and complex, probably due to synergistic or non-synergistic amplification of separate regions of 20q, involving multiple, independently amplified targets.

Induction of apoptosis in human choriocarcinoma cell lines by treatment with 3,4-dihydro-6-[4-(3,4-dimethoxybenzoyl)-1-piperazinyl]-2(1H)-quinolinone (vesnarinone).

Induction of apoptosis is an attractive strategy in cancer therapy but it clinical practice is not yet sufficient in choriocarcinoma. The quinolinone derivative, vesnarinone, is a novel inotropic agent used for treating congestive heart failure and may also have a potential anticancer activity. It induces apoptosis and differentiation in some tumor cell lines. We examined the antitumor effect of vesnarinone in eight cell lines established from human choriocarcinoma and hydatidiform mole using MTT assay and also analyzed the nuclear fragmentation of tumor cells by DNA electrophoresis assay. Vesnarinone inhibited the proliferation of choriocarcinoma cell lines in a dose-dependent manner and induced DNA fragmentation in cells. However, the BM cell line prepared by subcultivation from hydatidiform mole showed no growth suppression or DNA fragmentation in response to vesnarinone. On the other hand, PCR-SSCP analysis and direct DNA sequencing have shown that a human choriocarcinoma cell line, SCH, has a mutant p53 gene at codon 249. When SCH cells were treated with vesnarinone cellular proliferation was significantly inhibited. Vesnarinone suppressed the proliferation of all choriocarcinoma cell lines and induced apoptosis, regardless of the existence of p53 mutation. In addition, it has been found by RT-PCR that expression of c-Myc mRNA is upregulated by treating choriocarcinoma cells with vesnarinone. The finding suggests that vesnarinone might induce expression of c-Myc gene in choriocarcinoma cells, the product of which may be associated with the inhibition of cell growth and induce apoptosis. These results suggest that vesnarinone is a useful reagent for the treatment of choriocarcinoma.

Establishment and characterization of a new human cell line (EJ) derived from endometrial carcinoma.

We present a new cell line, EJ established from an invasive endometrioid adenocarcinoma of the uterine corpus in a 56-year-old patient. The cells show rapid growth in culture with a doubling time of 16 h and high migration activity. Monolayer-cultured cells were polygonal in shape showing a tendency to pile up without contact inhibition. Subcutaneous transplantation of the EJ cells into nude mice formed solid tumors that were histologically diagnosed as adenocarcinoma, whereas no metastasis was observed. Cultured EJ cells produced tissue polypeptide antigen (IPA). Genetic and molecular analyses revealed high telomerase activity but not estrogen receptor alpha expression. Using the DNA sequencing technique, we have screened EJ cells for p53 mutation in exon 5 to 8 but no mutation of p53 was observed. This cell line appears to represent the development of a more malignant clone with divergent receptor function and growth behavior, and provides us with an interesting new tool for the study of tumorigenesis in the human endometrium.

Dysgerminoma in a patient with a tumor of the neck. Empiric treatment of stage IV dysgerminoma.

The evolution of therapy for malignant ovarian germ cell tumors is one of the true success stories in oncology. Treatment outcome has improved greatly thanks to cisplatin-based combination chemotherapy. According to the well-established treatment guidelines for advanced cases, we treated a case of stage IV undifferentiated germ cell tumor in which we were able to preserve the patient's fertility. We concluded that the PEP regimen is an effective treatment for the patient with metastatic germ cell tumor.