Decreased expression of 14-3-3σ is predictive of poor prognosis for patients with human uterine papillary serous carcinoma.

Uterine papillary serous carcinoma (UPSC) morphologically resembles ovarian serous carcinoma and is categorized as a type II endometrial cancer. UPSC comprises about 10% of all types of endometrial cancer and has an aggressive clinical course and a poor prognosis. The 14-3-3σ gene was originally discovered as a p53-inducible gene; its expression is induced by DNA damage in a p53-dependent manner, which leads to G2 arrest and repair of damaged DNA. Moreover, it has been reported that expression of 14-3-3σ is frequently lost in various types of human cancer, including ovarian cancer. We therefore examined the association between 14-3-3σ expression determined by immunohistochemistry and clinical outcomes of 51 patients with UPSC. UPSC was considered positive for 14-3-3σ when > 30% of tumor cells were stained with a specific antibody. Of these patients, 29 (58.7%) showed positive immunoreactivity for 14-3-3σ and 22 (41.3%) had decreased 14-3-3σ staining. Decreased immunoreactivity for 14-3-3σ was associated with stage (P = 0.001) and lymphovascular space involvement (P = 0.005). Moreover, decreased 14-3-3σ expression was an independent risk factor for reduced overall survival (P = 0.0416) in multivariate analysis. Direct bisulfite sequencing was performed to evaluate the methylation status of the 27 CpG islands in the promoter region and first exon of the 14-3-3σ gene. These CpG islands were hypermethylated in 30% of 14-3-3σ-positive UPSC and 80% of 14-3-3σ-negative UPSC, although the difference was not statistically significant. These findings suggest that decreased expression of immunoreactive 14-3-3σ may be a predictor of poor prognosis in patients with UPSC.

Association of keap1 and nrf2 genetic mutations and polymorphisms with endometrioid endometrial adenocarcinoma survival.

OBJECTIVE: Dysregulation of Kelch-like erythroid cell-derived protein with CNC homology-associating protein (Keap)-nuclear factor E2-related factor 2 (Nrf2) homeostasis owing to oncogenic mutations or to endogenous alteration of protein expression levels is implicated in tumor resistance to adjuvant treatment. To understand the role of Keap1 and Nrf2 in endometrial cancer, we performed DNA sequencing of tumors and noted the relation of the DNA sequence with annotated clinicopathologic data. METHODS: We sequenced the Keap1 and Nrf2 genes in 105 tumor specimens. Associations of genetic mutations and polymorphisms with the patients' clinicopathologic characteristics were evaluated. RESULTS: We detected 9 patients with Keap1 mutations and 3 patients with Nrf2 mutations. No patient had both Keap1 and Nrf2 mutations. We found 2 single nucleotide polymorphisms within the coding region of Keap1 - rs1048290 (c. 1413C>G) and rs11545829 (c. 1611C>T) that displayed high heterogeneity in our cohort. The c. 1413C>G polymorphism is significantly associated with progression-free survival by multivariate analysis (hazard ratio, 0.16; 95% confidence interval, 0.036-0.69; P = 0.014). The presence of Keap1 or Nrf2 mutations and c. 1611C>T are not associated with the clinical outcome of the patients. CONCLUSIONS: Mutations impairing Keap1-Nrf2 interaction are relatively common in endometrial cancer (12 [11.4%] of 105). Keap1 single nucleotide polymorphism rs1048290 may be a novel independent prognostic marker for patients with endometrial cancer receiving adjuvant treatment. Therefore, genotyping patients for this Keap1 polymorphism will help identify patient subgroups more likely to benefit from standard adjuvant therapy.

Roles of intrinsic angiogenesis inhibitor, vasohibin, in cervical carcinomas.

The aim of the present study is to clarify the critical roles of vasohibin in cervical carcinomas. We investigated the expression ratios of vasohibin and vascular endothelial growth factor (VEGF) receptor-2 on endothelium and microvessel density, lymphatic vessel density (LVD) by immunohistochemistry. Sixty-one squamous cell carcinoma (SCC), 18 mucinous adenocarcinoma (Adenocarcinoma), 38 carcinoma in situ (CIS), and 35 normal cervical epithelium were collected. We investigated the expression of vasohibin and compared it with the expression of VEGF receptor-2 (VEGFR-2, KDR/flk-1), and CD34 in the stromal endothelium. Expression of VEGF was counted using the histological score (H score). D2-40 was used as a marker for lymphatic endothelial cells to investigate LVD. The microvessel density of the normal cervical epithelium was significantly lower than that of CIS, SCC, and Adenocarcinoma (P < 0.05). The expression ratio of vasohibin in the normal cervical epithelium was significantly lower than that of SCC and Adenocarcinoma (P < 0.05). The expression ratio of VEGFR-2 of the normal cervical epithelium was significantly lower than that of SCC and Adenocarcinoma (P < 0.05). The LVD of the normal cervical epithelium was significantly lower than that of CIS, SCC, and Adenocarcinoma (P < 0.05). For normal cervical epithelium, CIS, and SCC, there was a moderate correlation between the expression percentage of vasohibin and the expression percentage of VEGFR-2 (P < 0.05, r(2) = 0.3018). This is the first study to elucidate the correlation between the expression of vasohibin in the stromal endothelial cells and the expression of VEGFR-2 in human cervical carcinomas.

Vasohibin-1 in human breast carcinoma: a potential negative feedback regulator of angiogenesis.

Vasohibin-1 is a recently identified negative feedback inhibitor or suppressor of angiogenesis induced by vascular endothelial growth factor (VEGF)-A. The status of vasohibin-1 in human breast carcinoma has not been examined. We examined 151 breast specimens including 98 cases of invasive ductal carcinoma (IDC), 12 of ductal carcinoma in situ (DCIS), 16 of fibroadenoma (FA), six of inflammatory lesion, nine of fibrocystic change and seven of non-pathological breast tissue. We immunolocalized vasohibin-1 and compared its immunoreactivity to that of VEGF-A, basic fibroblastic growth factor (bFGF), VEGF receptor 2 (Flk-1), CD31, CD34 and Ki-67/MIB-1. The correlation of vasohibin-1 immunoreactivity with overall survival (OS), and disease-free survival (DFS) of the patients with breast carcinoma was also evaluated. In addition, we evaluated Ki-67 and CD31, and Ki-67 and vasohibin-1 double-immunostaining for further characterization of neovascularization. Vasohibin-1 was detected in endothelial cells of human breast and its immunodensity was significantly higher in IDC and inflammatory lesions than the other types (P<0.001). In addition, a significant positive correlation was detected between vasohibin-1 and VEGF-A, bFGF or Flk-1 (P<0.001). There was also positive associations between vasohibin-1 and OS (P=0.004) and between vasohibin-1 and DFS (P

Expression of vasohibin as a novel endothelium-derived angiogenesis inhibitor in endometrial cancer.

We have previously reported on vasohibin as a novel endothelium-derived vascular endothelial growth factor (VEGF)-inducible inhibitor of angiogenesis. The aim of our present study was to define the role of vasohibin in endometrioid endometrial adenocarcinoma. We collected 78 sections of endometrial carcinoma for assessment using immunohistochemistry. Twenty-seven were well differentiated (G1), 25 were moderately differentiated (G2), and 26 were poorly differentiated endometrioid adenocarcinomas (G3). We also included 12 sections of normal cyclic endometria, six of which were in the proliferative phase and six were in the secretory phase. We investigated the expression of vasohibin, and compared it to VEGF receptor-2 (VEGFR-2: KDR/flk-1), CD34, Ki-67, VEGF-A, and D2-40 (as a lymphatic vessel marker). We assessed the ratio of vasohibin- and VEGFR-2-positive vessels in the stroma of endometrial carcinoma. Immunohistochemical assessment was classified as negative or positive based on staining intensity. Vasohibin was selectively expressed on vascular endothelial cells in both cyclic endometria and endometrial carcinomas. Vasohibin was highly expressed in the normal functional endmetrium of the secretory phase, especially in the spiral artery, and was highly expressed in all grades of endometrioid adenocarcinomas. The stromal endothelial cells in G3 expressed vasohibin and VEGFR-2 more frequently than these in G1. In endometrioid adenocarcinomas, there was a significant correlation between the expression percentage of vasohibin and that of VEGFR-2 (P < 0.0001, r(2) = 0.591). This is the first study to elucidate the correlation between expression of vasohibin in the stromal endothelial cells and that of VEGFR-2 in human carcinomas.

A case of primary mucosa-associated lymphoid tissue lymphoma of the vagina.

We report the first case of primary mucosa-associated lymphoid tissue (MALT) lymphoma of the vagina, the diagnosis of which is supported by genetic and immunophenotypic studies. A 65-year-old, para 2 woman presented to our hospital in July 1997 with a history of prolonged vaginal discharge. Although cytologic examination suggested possible malignancy, a biopsy of the vaginal wall was diagnosed as chronic inflammation. In June 2000, she underwent gynecologic examination because of anuria. Excisional biopsy revealed subepithelial infiltration of atypical lymphoid cells that stained for CD20, CD79a, and BCL-2; stained weakly for IgM; and did not stain for CD3, CD5, CD7, CD10, CD56, CD23, and IgD, suggesting marginal zone B-cell lineage. Monoclonality was detected by Southern blot analysis, and this patient was finally diagnosed as having primary MALT lymphoma of the vagina. She received 3 cycles of chemotherapy (THP-COP) and concurrent radiation to the whole pelvis. The patient is alive and well 40 months after treatment. Because the vagina is one of the mucosa-associated tissues, MALT lymphoma, though rare, must be included in the differential diagnosis of the vaginal neoplasms.

The cul-de-sac packing method with a metreurynter in gynecologic gasless laparoscopy.

Laparoscopic surgery has inherent restrictions with respect to the operative field of view and the range of surgical manipulation. Of the two procedures which secure sufficient operative space, the operative view of the gasless method is inferior to that of a pneumoperitoneum. In order to gain greater surgical visualization in gynecological gasless laparoscopy, the authors devised the cul-de-sac packing method employing a metreurynter, an instrument familiar to obstetricians in Japan. A metreurynter was lead into the cul-de-sac, and was inflated with saline, which resulted in the adnexae being raised up. This method was performed in three patients whose preoperative diagnoses were unknown infertility, ovarian cyst, and ectopic pregnancy, respectively. In all cases this method was able to keep the bowels out of the cul-de-sac space. In the first case, we were able to perform a tubal patency test under tension-free conditions, while at the same time bilateral tubal information could be obtained in a single view. In the latter two cases the adnexal lesions were maintained at an inspectional position throughout the operation without the necessity of being held by forceps to prevent them from falling down into the cul-de-sac space. No complications occurred in our three cases. This method will not be useful for patients whose cul-de-sac space is closed due to adhesions. However, except in such cases, this technique supplies a good operative view while being simple, safe, and inexpensive. Furthermore, this method supports gentler and less traumatic manipulation throughout the operation.