Hiromitsu Yabushita

Vitamin D Receptor in Endometrial Carcinoma and the Differentiation-Inducing Effect of 1,25-Dihydroxyvitamin D3 on Endometrial Carcinoma Cell Lines

In view of the potential of 1,25‐dihydroxyvitamin D3 [1,25(OH)2D3] as a cell‐differentiation‐inducing agent in endometrial cancer, the localization of the vitamin D receptor (VDR) was examined immunohistochemically in 21 endometrial adenocarcinoma specimens, and the effect of 1,25(OH)2D3 on cell growth, as well as the phenotypic changes for cell maturation after treatment with 1,25(OH)2D3, was investigated in 2 endometrial carcinoma cell lines (AMEC‐1, RL95‐2). The VDR was detected in 14 of the 21 endometrial carcinoma specimens. The growth of RL95‐2 cells expressing VDR was inhibited to 44% when cultured with 50 nM 1,25(OH)2D3 for 6 days. In contrast, the growth of AMEC‐1 cells not expressing VDR was completely uninhibited even when cultured with 100 nM 1,25(OH)2D3 for 6 days. The RL95‐2 cells exposed to 50 nM 1,25(OH)2D3 for 6 days had an increasing expression for 52.5 kD or 45 kD cytokeratin polypeptide, and they became columnar with pronounced polarity and formed gland‐like structures when cultured in collagen gel. These results suggest that endometrial adenocarcinoma is a target for 1,25(OH)2D3, which appears to function as a cell‐differentiation‐inducing agent for the treatment of endometrial cancer.

Combination assay of CA125, TPA, IAP, CEA, and ferritin in serum for ovarian cancer☆

The levels of CA125, TPA, IAP, CEA, and ferritin in the serum were measured simultaneously in 68 healthy nonpregnant females and 133 patients with various gynecological diseases, and were subjected to statistical discriminant analysis for the diagnosis of ovarian cancer. The usefulness and the limits for diagnosis of various gynecological diseases were investigated for each tumor marker. Also, the diagnostic usefulness of the stepwise discriminant analysis employing the values of these five tumor markers in the serum in cases of ovarian cancer was compared with that of CA125 measurements alone. Because the frequency of cases with an elevated serum CA125 level increased more specifically in the ovarian cancer group than those of other tumor makers in the serum, this parameter was considered to be more useful for the diagnosis of ovarian cancer than the levels of the other tumor markers. The frequencies of cases with the elevated serum CA125 levels, however, also increased in the groups of patients with endometriosis and at an early stage of normal pregnancy more than in the group of healthy nonpregnant females. In the ovarian cancer patients, the discriminant analysis employing the values of CA125 and four other tumor markers in sera was more useful for early diagnosis, differential diagnosis, early detection of recurrences, and the determination of complete remission after therapy than the measurement of the serum CA125 level alone.

The Association of Transforming Growth Factor‐β1 with Myometrial Invasion of Endometrial Carcinomas through Effects on Matrix Metalloproteinase

Objective: The association of transforming growth factor‐β1 (TGF‐β1) with a matrix metalloproteinase (MMP) and a tissue inhibitor of metalloproteinase (TIMP), as well as myometrial invasion of endometrial cancer was studied.

Clear-cell adenocarcinoma of the uterine cervix in a 17-year-old adolescent

This report describes the case of the youngest Japanese person to be diagnosed with endocervical clearcell adenocarcinoma. In September 2005, a 17-year-old female adolescent visited a physician because of vaginal bleeding. A cervical tumor was discovered, and the patient was referred to our outpatient department. Vaginal examination showed a bleeding tumor approximately 1.5 cm in size protruding from the cervical os. The cytological finding of the uterine cervix was positive for malignancy, and the histological diagnosis by punch biopsy was clear-cell adenocarcinoma of the uterine cervix. A radical abdominal hysterectomy and pelvic lymphadenectomy were performed on October 30. Macroscopic findings showed a tumor, 1.5 cm in diameter, growing from the right side of the uterine cervix. The pathological diagnosis was clear-cell adenocarcinoma of the cervix (PT1b1NR0M0). The patient was discharged from our hospital without any adjuvant therapy. No signs of recurrence have been detected in the 2-year follow up.

Role of hypoxia-inducible factor-1α, carbonic anhydrase-IX, glucose transporter-1 and vascular endothelial growth factor associated with lymph node metastasis and recurrence in patients with locally advanced cervical cancer

The aim of the present study was to determine whether the expression of hypoxia-inducible factor-1α (HIF-1α), carbonic anhydrase-IX (CA-IX), glucose transporter-1 (GLUT-1) or vascular endothelial growth factor (VEGF) was associated with the clinicopathological characteristics, lymph node metastasis or progression-free survival of patients with cervical cancer. Tumor tissue samples were obtained from 54 cervical cancer patients who had undergone radical hysterectomy. The expression of HIF-1α, CA-IX, GLUT-1 and VEGF was analyzed by immunohistochemical staining. Of the 54 cases, 28 were positive for HIF-1α, 35 for CA-IX, 40 for GLUT-1 and 23 for VEGF. It was revealed that HIF-1α expression was correlated with tumor stage and histology, CA-IX expression with tumor stage, tumor size, lymph node metastasis and lymph-vascular space involvement, GLUT-1 expression with tumor stage and lymph-vascular space involvement, and VEGF expression with microvessel density. The multivariate regression analysis indicated that CA-IX expression and lymph-vascular space involvement were independent variables associated with lymph node metastasis. Progression-free survival was shorter for patients who were positive for CA-IX or VEGF expression than for those who were negative for CA-IX or VEGF expression. The progression-free survival of patients treated with radiotherapy or chemo-radiotherapy following radical hysterectomy was also shorter for patients with positive CA-IX expression. These findings suggest that CA-IX expression is a possible risk factor for lymph node metastasis and disease recurrence in locally advanced cervical cancer patients.

Growth potential of endometrial cancers assessed by a Ki-67 Ag/DNA dual-color flow cytometric assay

Ki-67 is a monoclonal antibody directed against a nuclear antigen present only in proliferating cells. To assess the growth potential of uterine endometrial cancer, the population of cells in proliferating cycle (%PC) was examined with Ki-67, using flow cytometry. The %PC of 27.18 +/- 12.00% in 22 endometrial cancers was significantly higher than the 14.5 +/- 5.94% found in 28 normal endometrial tissues. In premenopausal endometrial tissue, the %PC in the proliferatory phase was significantly higher than the %PC found in the secretory phase. In endometrial cancers, an increase of %PC was found in cases with deep myometrial invasion, and the %PC was elevated in groups containing histologically poorly differentiated types when compared to groups of well-differentiated and moderately differentiated types. Sorted cells reactive with Ki-67 antibody were large and had a high nuclear/cytoplasmic ratio. On the bases of these results, it was concluded that a Ki-67 Ag/DNA dual-color assay would be useful to examine the growth fraction in endometrial carcinoma and that an increased growth fraction was related to deep myometrial invasion or poorly differentiated types.

Clinicopathological Role of Serum-Derived Hyaluronan-Associated Protein (SHAP)-Hyaluronan Complex in Endometrial Cancer.

The role of hyaluronan (HA), serum-derived HA-associated protein (SHAP)-HA complex and hyaluronan synthase (HAS) in endometrial carcinomas was investigated. The relationship of metalloproteinase (MMP) and its inhibitor (TIMP) with HA and the SHAP-HA complex was also examined. The expression of HAS1 was related to the depth of myometrial invasion and lymph-vascular space involvement. The serum levels of HA, SHAP-HA complex, MMP-9, and TIMP-1 were increased in related with the depth of myometrial invasion, histological grade and lymph-vascular space involvement. They were also higher in the HAS1-positive group compared to -negative group. The serum concentrations of HA and SHAP-HA complex had a significant correlation with the MMP-9 and TIMP-1. The patients with elevated SHAP-HA complex had the shorter disease-free survival. The multivariate analysis revealed that the SHAP-HA complex was the independent variable for disease-free survival of endometrial cancer patients. In conclusion, the elevation of serum SHAP-HA complex depended on the HAS1 expression and the SHAP-HA complex is a useful marker to predict disease recurrence in endometrial cancer patients. The SHAP-HA complex may promote the lymph-vascular space involvement and the synthesis and activation of MMP-9 and TIMP-1 in the progression of endometrial cancer.

Detection of Chlamydia trachomatis infection with DNA extracted from formalin-fixed paraffin-embedded tissues.

It is a well-known fact that tubal stenosis and/or peritubal adhesion are often associated with Chlamydia trachomatis infection. Although tubal pregnancy may be attributed to this infection, there are only extremely rare cases in which the presence of C. trachomatis has been confirmed by immumo-histochemical examination on tissues isolated from patients with tubal pregnancy. We thus tried to confirm the presence of C. trachomatis infection by detecting DNA of the organism in tissues surgically isolated from patients with tubal pregnancy. Two detection methods, a ligase chain reaction (LCR) method and an immuno-histochemical staining which detects an antigen of C. trachomatis, were compared using paraffin-embedded tissue samples which were surgically isolated from patients with tubal pregnancy or hydrosalpinx. The LCR method was capable of detecting DNA of C. trachomatis in tissue samples in which the C. trachomatis-specific antigen could not be detected using immuno-histochemical staining. This was due to the fact that immuno-histochemical staining methods are applicable to the analysis of sequences the length of which range from 200 to 400 bp (base pairs), while the LCR method used here allows the analysis of sequences as small as 48 bp. This fact makes the LCR method a very convenient tool, as compared with immuno-histochemical methods, for analysis of the paraffin embedded tissue samples where by effects of formalin--used for fixation for pathologic diagnosis--the risk of fragmenting the DNA samples is relatively high. Presence of C. trachomatis DNA as detected by LCR method in surgically isolated samples from patients with tubal pregnancy supports the involvement of Chlamydia trachomatis infection in the occurrence of tubal pregnancy. Accordingly the LCR method is capable of detecting DNA of C. trachomatis in paraffin-embedded samples of tubal tissue in which presence of C. trachomatis could not be confirmed by an immuno-histochemical staining method.

Hyaluronan (HA) and serum-derived hyaluronan-associated protein (SHAP)-HA complex as predictive markers of cervical ripening in premature labor.

The purpose of this study is to investigate whether serum hyaluronan (HA) and serum-derived HA-associated proteins (SHAP)-HA complex predict cervical ripening and premature delivery. Sera were obtained from 64 women with normal pregnancies, 20 with full term delivery, and 13 with threatened premature labor. Concentrations of HA and SHAP-HA complex in serum were measured by sandwich ELISA. Serum concentrations of HA and SHAP-HA complex did not differ within first, second, and third trimester groups. The serum SHAP-HA complex was elevated in the full term labor group more than in the third trimester group; however, the concentrations of serum HA did not differ between both groups. The HA and SHAP-HA complex levels in sera were higher in the premature labor group than in the second trimester group. In the premature labor group, the SHAP-HA complex levels were higher in the cases with Bishop scores more than 4 points when compared with the cases with Bishop scores of 4 points or less. Increased levels of SHAP-HA complex in sera are possible predictive markers for cervical ripening in premature labor.

Clinicopathological roles of adiponectin and leptin receptors in endometrial carcinoma

To clarify the roles of adiponectin receptor (AdipoR) and leptin receptor (ObR) in endometrial carcinoma, the expression of AdipoR-1 and -2 and ObR in endometrial cancer was examined immunohistochemically, and correlations with clinicopathological implications were also analysed. Paraffin-embedded tissues were obtained from 77 patients with endometrial carcinoma and were stained immunohistochemically using antibodies against AdipoR-1, AdipoR-2 and ObR. AdipoR-1, AdipoR-2 and ObR were localised predominantly in the cell membrane and cytoplasm of tumour cells and normal endometrial cells. In 77 cases of endometrial cancer, positive expression was observed in 46 cases (59.7%) for AdipoR-1, 47 cases (61.0%) for AdipoR-2 and 33 cases (42.9%) for ObR. Expression of AdipoR-1 was observed most in stage I cases, G1 tumours, tumours with shallow myometrial invasion, tumours negative for lymphovascular space involvement, cases negative for adnexal invasion and cases with no lymph node metastasis. However, the expression of AdipoR-2 and ObR showed no correlation with any clinicopathological factors. Kaplan-Meier analyses revealed that progression-free and overall survival times were longer in cases with positive AdipoR-1 expression compared with negative AdipoR-1 expression. Poor expression of AdipoR-1, thus, appears to be associated with tumour grade, myometrial invasion, adnexal invasion, lymph-vascular space involvement and lymph node metastasis, as well as poor prognosis, in endometrial cancer.