Morio Sawada

Different expression patterns of KIT, EGFR, and HER-2 (c-erbB-2) oncoproteins between epithelial and mesenchymal components in uterine carcinosarcoma

Uterine carcinosarcoma histologically comprises the components of epithelial and mesenchymal malignancies, and is known to be clinically highly aggressive. To reveal the significance of the expression of tyrosine‐kinase‐receptor‐type oncoproteins in this tumor type, the incidence and distribution of the KIT, EGFR, and HER‐2 (c‐erbB‐2) oncoproteins were immunohistochemically examined in 16 surgically resected cases. For 6 cases, the EGFR and HER‐2 amplifications were also examined by fluorescence in situ hybridization (FISH). In the epithelial component, overexpressions of KIT, EGFR, and HER‐2 were detected in 4 (25%), 5 (31%), and 9 (56%) cases, respectively, whereas these overexpressions in the mesenchymal component were detected in 6 (38%), 8 (50%), and 1 (6%) cases, respectively. KIT and EGFR were co‐overexpressed in the mesenchymal component of 4 cases and in the epithelial component of 2 cases. However, HER‐2 overexpression was mostly detected in the epithelial component only, and tended to occur independently of KIT and/or EGFR overexpression. By FISH, one of the 4 cases with HER‐2 overexpression showed low‐level gene amplification. In two cases with EGFR overexpression, the gain of EGFR alleles and/or polyploidization of chromosome 7 had occurred. The expression patterns of KIT, EGFR, and HER‐2 differed between the epithelial and mesenchymal components, and the regulation of their expression appeared important in the acquisition of mesenchymal metaplasia in uterine carcinosarcoma. Structural and/or numerical alterations of chromosomes might be in part involved in EGFR and/or HER‐2 overexpression in this tumor type.

Radical hysterectomy for FIGO stage IIB cervical cancer: clinicopathological characteristics and prognostic evaluation.

OBJECTIVE To clarify the clinicopathological features and prognostic factors of patients with FIGO stage IIB cervical cancer who were treated with radical hysterectomy. METHODS One hundred thirty-nine FIGO stage IIB patients with squamous or adenosquamous cell carcinoma (median age, 51 years) who were treated with primary radical hysterectomy were examined retrospectively. Sixty-six FIGO stage IIB patients who were treated with primary radiotherapy (median age, 70 years) were included for comparison of survival. RESULTS Fifty percent (70/139) of the patients had pathological parametrial involvement. Among them, the positive rate of pelvic lymph nodes was 71% (50/70). Ninety-nine percent (138/139) of the tumors were completely removed, and the pelvic control rate was 88%. Major complications requiring surgery were found in 2.9% (4/139). Significant differences in survival were found among patients in subgroups according to pathological parametrial involvement, pelvic lymph node status, tumor size, lymph-vascular space invasion, and depth of myometrial invasion (log-rank test, P<0.05). Of these, the Cox proportional-hazard model revealed that parametrial involvement (P=0.001, 95% CI 1.992-6.297) and lymph node metastasis (P=0.042, 95% CI 1.023-3.298) were independent prognostic factors. The 5-year survival rate and relapse-free survival at 36 months were 69% and 72% among the radical hysterectomy group, and 69% and 75% among the radiotherapy group. The Cox model adjusted for age showed no significant differences in survival and relapse-free survival between these two groups. CONCLUSION Pathological parametrial involvement and positive nodes were prognostic factors for surgically treated patients with FIGO stage IIB cervical cancer.

Neoadjuvant weekly carboplatin and paclitaxel followed by radical hysterectomy for locally advanced cervical cancer: long-term results.

Introduction: To determine the long-term effect of neoadjuvant chemotherapy with paclitaxel and carboplatin on a weekly schedule followed by radical surgery for patients with locally advanced cervical cancer. Materials and Methods: Thirty patients with stage IB2 to IIIB uterine cervical cancer were treated with paclitaxel (60 mg/m2) and carboplatin (area under the curve, 2-an area under the time-concentration curve of 2 mg × min/mL based on creatinine clearance) every week for 6 cycles. A radical hysterectomy was performed 6 days after the final administration of neoadjuvant chemotherapy. The patients were followed up, and 5-year progression-free survival (PFS) and overall survival (OS) were evaluated. Results: Of 30 patients, 28 were followed up. The median follow-up period was 55.6 months (range, 26-83 months). An objective response (complete response + partial response) to the treatment was observed in 26 patients (87%; 95% confidence interval, 70%-95%). Two had complete response, 4 had stable disease, and the remaining patients had partial response; progressive disease was not seen in this study. A radical hysterectomy was performed in 28 patients without delay. Thirteen patients with high-risk factors received radiotherapy after surgery. The 5-year PFS and OS rates were 78.6% and 81.8%, respectively. The 5-year PFS and OS for patients with stage IB2 to IIB cervical cancer were 79.2% and 83.1%, respectively, which were comparable with those in the concurrent chemoradiation therapy study previously reported. There was no significant correlation in survival between preoperative staging and cell type, whereas larger initial tumor size and lymph node metastasis tended to be negatively correlated with survival. Conclusions: Neoadjuvant chemotherapy with paclitaxel and carboplatin on a weekly schedule followed by radical surgery for patients with locally advanced cervical cancer is a promising mode of therapy that may improve the prognosis. It would be worthwhile to conduct larger-scale trials for comparison with the results of the chemoradiation therapy study.

Estrogen-related receptor α expression and function are associated with vascular endothelial growth factor in human cervical cancer.

Introduction: Estrogen-related receptor &agr; (ERR&agr;), one of orphan nuclear receptors with an unknown ligand, is expressed in various types of cancer. Increased ERR&agr; levels are associated with a higher risk of recurrence and poor clinical outcome in breast cancer, suggesting that ERR&agr; could be a negative prognostic factor. Recently, it has been suggested that vascular endothelial growth factor (VEGF) could be one of the transcriptional targets of ERR&agr; in breast cancer. Here, we examined the expression of ERR&agr; and the association of ERR&agr; with VEGF in uterine cervical cancer cells and tissues. Methods: We evaluated the expression of ERR&agr; and VEGF by immunohistologic analysis using specimens from 40 patients with invasive cervical cancer. We also evaluated the VEGF promoter activity of ERR&agr; in cervical cancer cell lines by transfection and luciferase assay. We overexpressed or knocked down ERR&agr; and examined VEGF expression by real-time polymerase chain reaction. Finally, cell proliferation assay was performed to examine whether ERR&agr; affects tumor growth in cervical cancer. Results: Immunohistologic analysis demonstrated that ERR&agr; expression in cervical cancer tissues was higher than that in noncancerous tissues and that there was a positive association between ERR&agr; and VEGF expression in cancer tissues (P < 0.05). We showed that ERR&agr; stimulated the VEGF promoter activity in cervical cancer cell lines. We further showed the overexpression and knockdown of ERR&agr;-regulated VEGF expression level by real-time polymerase chain reaction. Moreover, we showed that ERR&agr; and VEGF knockdown by small interfering RNA or an inverse agonist of ERR&agr;, XCT 790, could suppress cell growth compared with control cells in cervical cancer. Conclusions: We have provided compelling evidence that ERR&agr; affects VEGF expression and tumor growth in cervical cancer. These results justify further investigation into the use of ERR&agr; as a therapeutic target for patients with uterine cervical cancer.

A case of vaginal clear cell adenocarcinoma complicated with congenital anomalies of the genitourinary tract and metanephric remnant without prenatal diethylstilbestrol exposure

Vaginal clear cell adenocarcinoma (CCA) is well known to be associated with prenatal diethylstilbestrol exposure. We present a vaginal CCA with congenital anomalies of the genitourinary tract without prenatal diethylstilbestrol exposure. A 54‐year‐old woman complained of a 3‐month history of genital bleeding. The examination revealed CCA at the anterior vagina and congenital anomalies. An anterior pelvic exenteration was performed. Macroscopically, bicornuate uterus, vaginal septum and left ureteral agenesis were found. Microscopically, vaginal CCA coexisted with adenosis and both metanephric and mesonephric remnants. The vaginal CCA was supposed to derive from coexisting adenosis. The adenosis was also supposed to occur as a congenital basis, together with genitourinary tract anomalies. Relations between congenital anomalies of the genitourinary tract and vaginal adenocarcinoma were suspected, resultantly.

Loss of AF-6/afadin induces cell invasion, suppresses the formation of glandular structures and might be a predictive marker of resistance to chemotherapy in endometrial cancer

BackgroundAF-6/afadin plays an important role in the formation of adherence junctions. In breast and colon cancer, loss of AF-6/afadin induces cell migration and cell invasion. We aimed to elucidate the role of AF-6/afadin in human endometrial cancer.MethodsMorphology and AF-6/afadin expression in endometrial cancer cell lines was investigated by 3-dimensional culture. We used Matrigel invasion assay to demonstrate AF-6/afadin knockdown induced invasive capability. Cell proliferation assay was performed to estimate chemoresistance to doxorubicin, paclitaxel and cisplatin induced by AF-6/afadin knockdown. The associations between AF-6/afadin expression and clinicopathological status were determined by immunohistochemical analysis in endometrial cancer tissues. Informed consent was obtained from all patients before the study.ResultsThe majority of cell clumps in 3-dimensional cultures of Ishikawa cells that strongly expressed AF-6/afadin showed round gland-like structures. In contrast, the cell clumps in 3-dimensional cultures of HEC1A and AN3CA cells—both weakly expressing AF-6/afadin—showed irregular gland-like structures and disorganized colonies with no gland-like structures, respectively. AF-6/afadin knockdown resulted in reduced number of gland-like structures in 3-dimensional cultures and enhancement of cell invasion and phosphorylation of ERK1/2 and Src in the highly AF-6/afadin-expressing endometrial cancer cell line. Inhibitors of MAPK/ERK kinase (MEK) (U0126) and Src (SU6656) suppressed the AF-6/afadin knockdown-induced invasive capability. AF-6/afadin knockdown induced chemoresistance to doxorubicin, paclitaxel and cisplatin in Ishikawa cells, not in HEC1A. Immunohistochemical analysis showed that AF-6/afadin expression was significantly associated with myometrial invasion and high histological grade.ConclusionsAF-6/afadin regulates cell morphology and invasiveness. Invasive capability is partly regulated through the ERK and Src pathway. The inhibitors to these pathways might be molecular-targeted drugs which suppress myometrial invasion in endometrial cancer. AF-6/afadin could be a useful selection marker for fertility-sparing therapy for patients with atypical hyperplasia or grade 1 endometrioid adenocarcinoma with no myometrial invasion. AF-6/afadin knockdown induced chemoresistance especially to cisplatin. Therefore, loss of AF-6/afadin might be a predictive marker of chemoresistance to cisplatin.

Leprdb/db Mice with Senescence Marker Protein-30 Knockout (Leprdb/dbSmp30Y/−) Exhibit Increases in Small Dense-LDL and Severe Fatty Liver Despite Being Fed a Standard Diet

Background/Aims The senescence marker protein-30 (SMP30) is a 34 kDa protein originally identified in rat liver that shows decreased levels with age. Several functional studies using SMP30 knockout (Smp30Y/−) mice established that SMP30 functions as an antioxidant and protects against apoptosis. To address the potential role of SMP30 in nonalcoholic fatty liver disease (NAFLD) pathogenesis, we established Smp30Y/− mice on a Leprdb/db background (Leprdb/dbSmp30Y/− mice). Research Design/Principal Findings Male Leprdb/dbSmp30Y/− mice were fed a standard diet (340 kcal/100 g, fat 5.6%) for 16 weeks whereupon the lipid/lipoprotein profiles, hepatic expression of genes related to lipid metabolism and endoplasmic reticulum stress markers were analyzed by HPLC, quantitative RT-PCR and western blotting, respectively. Changes in the liver at a histological level were also investigated. The amount of SMP30 mRNA and protein in livers was decreased in Leprdb/dbSmp30Y/+ mice compared with Leprdb/+Smp30Y/+ mice. Compared with Leprdb/dbSmp30Y/+ mice, 24 week old Leprdb/dbSmp30Y/− mice showed: i) increased small dense LDL-cho and decreased HDL-cho levels; ii) fatty liver accompanied by numerous inflammatory cells and increased oxidative stress; iii) decreased mRNA expression of genes involved in fatty acid oxidation (PPARα) and lipoprotein uptake (LDLR and VLDLR) but increased CD36 levels; and iv) increased endoplasmic reticulum stress. Conclusion Our data strongly suggest that SMP30 is closely associated with NAFLD pathogenesis, and might be a possible therapeutic target for NAFLD.

Primary extraskeletal myxoid chondrosarcoma of the vulva

Primary extraskeletal myxoid chondrosarcoma (EMC) of the vulva is extremely rare. There is little available information about the biological behavior and treatment strategy for primary EMC of the vulva. We report a rare case of primary EMC of the vulva treated surgically. A 24‐year‐old Japanese woman had demonstrated a small and elastic mass of the vulva and underwent enucleation of the mass at a previous hospital, but a definitive histopathological diagnosis was not obtained. Therefore, the patient was referred to our hospital for further evaluation and treatment. We histopathologically diagnosed the tumor as primary EMC of the vulva and performed vulvectomy with vulvoperineal reconstruction. Microscopic examination of the resected specimens demonstrated residual tumor nodules of EMC. However, there were no viable tumor cells at the surgical margin. Approximately two years after wide local excision was performed, the patient is doing well and there is no apparent recurrence of EMC.

Prognostic impact of the history of breast cancer and of hormone therapy in uterine carcinosarcoma.

Objective Recent studies reveal an association between hormone therapy for breast cancer (BC), such as tamoxifen (TAM) and toremifene (TOR), and uterine carcinosarcoma (UCS). The aim of this study was to investigate the characteristics and prognosis of patients with UCS after BC and hormone therapy. Methods Between January 1997 and December 2007, we treated 51 patients with UCS. The medical records of these patients were reviewed, and factors that influenced their survival were retrospectively analyzed using univariate and multivariate analyses. Results Ten (19.6%) of the 51 patients had a history of BC; 6 (11.8%) had received hormone therapy with TAM or TOR. The characteristics of the patients with UCS were similar regardless of whether they had a history of BC or hormone therapy. On univariate analysis, age greater than 56 years, elevated serum lactate dehydrogenase levels, residual tumors, FIGO (International Federation of Gynecology and Obstetrics) stage higher than stage IIIa, and non–endometrioid carcinomatous components were identified as prognostic factors. On multivariate analysis, in addition to residual tumors, FIGO stage higher than stage IIIa, and non–endometrioid carcinomatous components, a history of BC (relative risk, 0.14), a history of TAM use (relative risk, 15.9), and a history of TOR use (relative risk, 16.9) were also identified as independently significant prognostic factors. Conclusions Our data suggest that a history of BC and hormone therapy for BC is a risk factor for developing UCS without obvious impacts on the characteristics of UCS. Both of these factors had statistically significant impacts on the prognosis of patients with UCS. Further studies are necessary to clarify and validate these associations.

Estrogen-related receptor-γ regulates estrogen receptor-α responsiveness in uterine endometrial cancer.

Objective Estrogen-related receptors (ERRs) are orphan nuclear receptors that modulate the estrogen receptor (ER)-mediated pathway and play roles in the regulation of breast and prostate cancer cell growth. However, the significance of the localization and the function of ERRs in uterine endometrial cancer remain unclear. We aimed to measure the expression of ERR&ggr; and determine its association with the ER-mediated pathway in human endometrial cancer. Methods Proliferation, luciferase, and quantitative polymerase chain reaction assays were performed in ER&agr;-positive (Ishikawa) and ER&agr;-negative (HEC1A) endometrial cancer cell lines. The association between ERR&ggr; and ER&agr; expressions was determined by immunohistochemical analysis in uterine endometrial cancer tissues. Results Estrogen-induced estrogen response element transcriptional activity was repressed by ERR&ggr; in ER&agr;-positive cells but was stimulated by ERR&ggr; in ER&agr;-negative cells. The stable overexpression of ERR&ggr; regulated the in vitro cell growth in the ER&agr;-positive and ER&agr;-negative endometrial cancer cell lines. A selective ERR&ggr; agonist, DY131, inhibited the growth of the ER&agr;-positive endometrial cancer cells but promoted that of the ER&agr;-negative cancer cells. Furthermore, we found that ERR&ggr; is expressed in the nuclei of human uterine endometrial cancer tissues. Estrogen-related receptor &ggr; was not associated with pathological parameters such as the International Federation of Gynecology and Obstetrics stage and histological type. The uterine endometrial cancer tissues with ERR&ggr;-positive/ER&agr;-negative status may have a significantly poor prognosis. Conclusions The relationship between ERR&ggr; and ER&agr; status could be a predictive marker for the treatment of uterine endometrial cancer, which provides an impetus for the identification of ligands for nuclear orphan receptor ERR&ggr;.