Analysis of E6 variants of human papillomavirus type 33, 52 and 58 in Japanese women with cervical intraepithelial neoplasia/cervical cancer in relation to their oncogenic potential.

The variation of the E6 region of human papillomavirus type 16 (HPV16) is associated with a high risk for cervical carcinogenesis. To see whether the same is the case with HPV33, 52 and 58, known to have high homology with HPV16, we analyzed the E6 sequence variation of these HPVs in 107 Japanese women with cervical intraepithelial neoplasia (CIN) or invasive cervical cancer (ICC): 20 HPV33-positive, 46 HPV52-positive and 41 HPV58-positive cases. HPV33 variants were more frequently observed in CINs I/II than in CIN III/ICCs (71% (5/7) versus 15% (2/13), P=0.02). In HPV52-positive cases, a single E6 variant was detected in 98% of the cases, whereas the prototype accounted for 98% of HPV58-positive cases. In summary, the distribution of E6 variants is different among HPV types tested, suggesting a link between E6 variation and oncogenic potential being type-specific.

Effects of toxic substances on female reproduction.

Successful reproduction requires a complex series of interdependent physiological, cellular and molecular events. In the female many of these interdependent events are vulnerable to interruption by xenobiotic compounds. The physiological steps in the female reproductive cycle are reviewed. Selected xenobiotics which interrupt this cycle are presented and their mechanisms and site of adverse effects are discussed. Finally, a more detailed discussion of chemically induced ovarian failure in the human and an experimental animal model system is presented.

Immunohistochemical characterization of undifferentiated carcinomas of the ovary.

Immunohistochemical characteristics of undifferentiated carcinomas of the ovary were examined using formalin-fixed, paraffin-embedded tissues with an avidin-biotin staining approach. Eight cases were collected from the pathology files of our Institute from a total of 214 recorded malignant ovarian tumors. For immunostaining, antibodies reacting with epithelial membrane antigen (EMA), pankeratin, vimentin, CA 125, CA 19-9, carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), alpha-1-antitrypsin (AT), epidermal growth factor receptor (EGFR), c-erbB-2, bcl-2 and p53 proteins were used. All the cases examined were positive for EMA and pankeratin, specific markers for epithelial tumors, negative for the non-epithelial tumor marker, vimentin, and also positive for EGFR. Interestingly, only one case was positive for CA 125, despite it being one of the commonest reported indicators of ovarian cancer. CA 19-9 was positive in 7 cases, CEA in 5, AFP in 2, AT in 6 and c-erbB-2 protein in 4. Two cases were positive for p53 protein, and in 1 of these positive staining for bcl-2 was also observed. These results indicate that the epithelial nature is well preserved in undifferentiated ovarian carcinomas, although consistently positive reactions were not observed within the cases for some antigens. They further clearly show that a negative signal for CA 125 can not be considered to exclude the possibility of a primary ovarian tumor.

Occurrence of apoptotic DNA fragmentation in quiescent and proliferating cells in human endometrial adenocarcinoma tissues and the influence of apoptosis-suppressing effects of bcl-2 products.

In situ estimation of DNA fragmentation by the nick end labelling (NEL) method, and immunohistochemical examination of Ki-67 proliferative antigen and bcl-2 products in human endometrial adenocarcinoma tissues were performed to provide answers to the following two questions; a) does apoptotic DNA fragmentation occur specifically in quiescent cells or in proliferating cells or randomly in both?, b) does the bcl-2 product exert its apoptosis-suppressing effects differentially on carcinoma cells depending on their cell cycle condition?. Serial sections, one micrometer in thickness, from formalin-fixed and paraffinembedded tissues of 9 cases of human endometrial adenocarcinoma were examined. Apoptotic DNA fragmentation was observed in both quiescent (Ki-67 negative) and proliferating (Ki-67 positive) cells. Bcl-2 product-positive tumor cell islands tended to be NEL negative, although a few but non-negligible number of carcinoma cells, including both Ki-67 positive and negative ones, were NEL positive. These results indicate that, at least in human endometrial adenocarcinomas, apoptotic DNA fragmentation and bcl-2 product-independent (DNA) fragmentation occurs non-specifically with respect to the cell proliferation status. Further, the results suggest an altered regulation of cell death processes in human solid tumor tissue in vivo.

Inverse correlation between bcl-2 expression and cell growth fraction in human endometrial adenocarcinoma tissue.

Correlation between expression status of bcl-2 products and cell growth fraction (estimated by immunostaining for Ki-67 antigen) was analyzed in human endometrial adenocarcinoma tissue in situ. For this, serial sections, 2 micrometers thick, from formalin-fixed and paraffin-embedded tissue samples of 10 cases of the carcinoma were examined. Nine out of 10 carcinomas contained both bcl-2 positive and negative nests when examined immunohistochemically. In the remaining one case, no positive reaction for bcl-2 was observed. In general, bcl-2 positive nests tended to contain quiescent (Ki-67 negative) carcinoma cells, and bcl-2 negative nests, on the contrary, a large fraction of proliferating (Ki-67 positive) cells. However, this correlation was not strict, and in a few nests, the level of growth fraction was observed to be similar irrespective of bcl-2 positivity. These results show the complex function(s) of bcl-2 products, when considering their apoptosis-suppressing effects, cell cycle dependence and influence on cell proliferation status. Further, the results suggest an altered regulation of oncogene products in human solid tumor tissue in vivo.

Evaluation of apoptosis in human endometrial adenocarcinoma: comparison of nick end labeling and Le(y) antigen immunostaining method.

In situ estimation of DNA fragmentation by the nick end labeling method and immunohistochemical detection of cell surface carbohydrate Le(y) were performed to establish correlation between the results of these two apoptosis-detecting techniques in human solid tumor tissues. Formalin-fixed and paraffin-embedded tissue samples from 10 cases of human endometrial adenocarcinoma were examined and compared by using the two techniques. A substantial fraction of carcinoma cells was found to be definitely stained by both methods, although Le(y) expression did not seem to be positively correlated with DNA fragmentation in the carcinoma tissue. These results suggest that Le(y) expression does not necessarily reflect apoptotic DNA fragmentation, at least in human endometrial adenocarcinomas, and simultaneous application of DNA nick end labeling and Le(y) immunostaining is necessary to show the relevant signs of apoptosis in histologic slides, especially gynecological cancers.

Expression of bcl-2 and apoptotic DNA fragmentation in human endometrial adenocarcinoma cells.

Immunohistochemical detection of bcl-2 products and in situ estimation of DNA fragmentation during apoptosis by the nick and labelling method were performed to establish any correlation between these two apparently opposing processes in human solid tumors. Serial sections, one micrometer in thickness, from formalin-fixed and paraffin-embedded tissues of 10 cases of human endometrial adenocarcinoma were examined using the two techniques. A significant fraction of carcinoma cells were found to be stained by both methods, although there existed a general tendency for bcl-2 expression to be negatively correlated with DNA fragmentation in carcinoma tissue. The results thus suggest that bcl-2 expression does not always block apoptosis, and that simultaneous application of bcl-2 immunostaining and DNA nick end labeling is useful for showing the complex nature of apoptosis in histologic slides, especially in gynecological cancers.

Tamoxifen mediated human endometrial carcinogenesis may not involve estrogenic pathways: a preliminary note.

Seven cases of endometrial adenocarcinoma patients who had experienced long-term tamoxifen treatments as adjuvant therapy of breast carcinoma, were investigated with respect to estrogen receptor (ER) status. Four cases of endometrial adenocarcinoma without tamoxifen treatment but with a previous history of breast carcinoma were investigated for comparison. One of the 7 and two of the 4 cases were positive for ER immunohistochemically. Thus, the frequency of ER positivity in secondary endometrial adenocarcinoma seemed to be at random among tamoxifen-treated and non-treated breast cancer patients. These results suggest that tamoxifen-mediated human endometrial carcinogenesis may not involve estrogenic pathway(s) but may involve other carcinogenic mechanisms such as DNA adduct formation as shown in rat liver tumorigenesis.

Immunohistochemical detection of ras p21 oncoprotein in undifferentiated and well-differentiated epithelial carcinomas of the human ovary.

Expression of ras p21 oncoproteins in human ovarian carcinomas was examined immunohistochemically by using a monoclonal antibody(clone RAS 10) with respect to the degree of their histological differentiation. To achieve this, the intensity of staining for the protein was compared between undifferentiated and well differentiated carcinomas, i.e. extreme subtypes of common epithelial carcinomas. The former was composed of 8 "solid" carcinomas and the latter, 11 serous, 8 mucinous, 4 endometrioid and 4 clear cell carcinomas. All the cases examined, including both undifferentiated and well-differentiated carcinomas, showed a positive reaction to this antibody. Staining intensity and the number of positive cells somewhat varied among the cases. Additionally, 2 cases of ovarian epithelial tumors of low malignant potential (I,MP) were stained with this antibody. Both the cases were positive, but the number of positive cells seemed to be rather less than that found in the carcinoma groups. Thus, no differences in ras p21 expression were observed between the cases examined in spite of the differences in the degree of differentiation of the epithelial ovarian carcinomas. However, the possibility remained that the number of positive cells could be an indicator of malignant potential, enabling us to distinguish LMPs from carcinomas.

Analysis of initial failure site and spread pattern in endometrial carcinoma: a Japanese experience.

Kasamatsu T, Shiromizu K, Takahashi M, Matsumoto K, Shirai T. Analysis of initial failure site and spread pattern in endometrial carcinoma: a Japanese experience. This retrospective study was undertaken in an attempt to identify initial failure sites and spread patterns in patients with endometrial carcinoma in Japan. A retrospective clinicopathologic review of 272 patients treated from 1983 to 1994 at Saitama Cancer Center was performed. Patients underwent total hysterectomy, bilateral salpingo-oophorectomy and pelvic lymphadenectomy. Postoperative external radiation was given to the patients with deep myometrial invasion, high grade tumor, and/or lymph node metastasis. Following surgery, chemotherapy was given to the patients with extrapelvic metastasis. Of the 272 patients, 262 had no residual disease after initial treatment and 10 had confirmed residual disease. Of the 262 patients, 32 (12.2%) suffered recurrence. The recurrence rates for stage I, II, III, and IV were 5.6% (10/178), 5.7% (2/35), 35.3% (18/51), and 100% (2/2), respectively. Of the 32 patients who failed, 6 (18.8%) experienced local failure, 13 (40.6%) had distant failure without peritoneal spread, and 13 (40.6%) had distant failure with peritoneal spread. In distant failure, the incidence of peritoneal spread was highest (50.0%, 13/26), closely followed by that of pulmonary metastasis (46.2%, 12/26). Furthermore, of those patients with residual disease, peritoneal spread was found in 80% (8 of 10). Five of the six patients (83.3%) with local failure survive, but all patients with peritoneal spread have died. Peritoneal dissemination is an important failure pattern and should be considered a top priority in an attempt to improve survival in patients with endometrial carcinoma.

Intramural carcinomas of the uterine corpus: a clinicopathological study.

Cases in which no sizable amount of tumor was observed in the uterine cavity and the main foci of the carcinoma were exclusively confined to the myometrium, were studied pathologically in a series of carcinomas of the uterine corpus. Ten out of 350 carcinomas of the uterine corpus proper fell into this category, and one case of suspicious metastatic corpus tumor from the ovary was also considered. The growth pattern of these tumors was classified into following 6 groups: (a) growth confined to the myometrium, (b) nodular growth in the myometrium with minimal endometrial involvement, (c) scattered (scirrhous) growth in the myometrium with marked shedding of the endometrium, (d) almost complete replacement of the myometrium by carcinoma with endometrial shedding (e) extreme subtype of usual endophytic growth pattern and (f) diffuse, intra-lymphatic spread of secondary tumor. One, 2, 3, 1, 3, and 1 cases of tumors of group, (a)-(f), were observed respectively in our pathological files. A spectrum of initial symptoms, diagnostic procedures, histology and histogenesis of the tumor was described in this report.

Malignant squamous cell elements in the ovarian cancer: report of 5 cases and review of the literature.

Five cases of ovarian cancer are presented in which malignant squamous cell elements were present. Three of the cases were adenosquamous carcinoma without composite endometriosis, mature cystic teratoma or with mixed mesodermal tumor. The other two cases were squamous cell carcinoma which arose in mature cystic teratoma, one of them being in situ carcinoma. Among cases of adenosquamous carcinoma, one composed predominantly of squamous cell element and foci of endometrioid adenocarcinoma was found. In this case, transitional figures between the two components were found. In other cases of adenosquamous carcinoma, components of adeno- and squamous cell carcinoma were intermingled in each carcinoma cell nests. Thus a large variety of histogenesis and microscopic figures were seen in malignant squamous cell elements in the ovarian cancer.

Cytopathological features of undifferentiated carcinoma of the ovary.

Cytopathological characteristics of undifferentiated carcinoma of the ovary were examined and reviewed. Seven cases of undifferentiated carcinoma were encountered among 202 malignant ovarian neoplasms in the Saitama Cancer Center. Histologically, two cases contained a minor component of usual Müllerian carcinoma and five others were purely of solid architecture without differentiation. Nevertheless, six cases, four of which were totally undifferentiated histologically, showed cytological features characteristic of adenocarcinoma. The results indicate that even carcinomas histologically diagnosed as undifferentiated may have cytological features of glandular differentiation, which may be Müllerian in origin.

Fine needle aspiration cytology of adenocarcinoma of the rectovaginal septum. A case report.

BACKGROUND: Adenocarcinoma arising in the rectovaginal septum is exceedingly rare and is difficult to diagnose by pathologic examination prior to surgery because of the anatomic position of the tumor. CASE: A 42-year-old woman presumed to have adenocarcinoma of the rectovaginal septum underwent fine needle aspiration for diagnosis. Although a previously performed biopsy from the posterior vaginal fornix was unsuccessful, fine needle aspiration cytology via the posterior vaginal wall detected adenocarcinoma cells. The cell clusters were composed of cells with enlarged and hyperchromatic nuclei. The nuclei themselves demonstrated round and/or irregular morphologic patterns, with high nuclear/cytoplasmic ratios, and often contained an enlarged, round nucleolus and sometimes multiple ones in a single nucleus. Aniso-nucleosis was severe, and the chromatin patterns ranged from coarse to finely granular. The cytoplasm was narrow and lightly stained. Following fine needle aspiration, the patient underwent posterior exenteration on the basis of the cytologic diagnosis. CONCLUSION: Fine needle aspiration cytology was useful in establishing the preoperative diagnosis of adenocarcinoma of the rectovaginal septum, and curative exenterative surgery could be then performed. To our knowledge, this is the first report of fine needle aspiration cytology of adenocarcinoma at this location.

Endometrial adenocarcinoma with diffused/scattered "intramural" spreads: report of a case and review of the literature.

A case is reported of endometrial adenocarcinoma of the uterus in an 85-year-old patient with an unusual spreading pattern. On macroscopic examination, only a tiny exophytic tumor was found in the uterine cavity, while microscopic examination demonstrated a scattered (scirrhous) spread of the carcinoma cells throughout the myometrium. The tumor occupied about half of the upper uterine corpus. The intramural spread of the tumor could not be seen at the time of macroscopic examination of the uterine cut surface. The tumor cells were attached closely to the serosal membrane, and metastasis to the left ovary was found. Intraoperative cytology detected malignant cells in the ascites. We present here this unusual type of endometrial carcinoma and review our previous report which dealt with pure "intramural carcinomas of the uterine corpus".

Pathological aspects of normal-sized ovarian carcinoma.

Twenty cases of ovarian carcinoma with normal-sized ovaries were examined histologically. Fifteen of them had metastatic disease of the peritoneal cavity and three of the remaining showed uterine involvement. Only in one case was the tumor confined to the ovary, but this patient had experienced systemic chemotherapy before surgery. The remaining case had metastasis of the stomach. Thus, "normal-sized" ovarian carcinoma has a great tendency to spread externally. Histology of the carcinoma was that of common epithelial carcinoma of the ovary with variable degrees of differentiation except for three cases of apparent ovarian metastasis. No cases of definite mesothelioma with ovarian involvement were encountered. A search for determinants of the peritoneal spread was performed using immunohistochemistry for p53 and bcl-2 proteins. However, no differences were found in the antigen expression between the group with and without peritoneal metastasis.

Cell dynamics of undifferentiated carcinomas of the ovary. Immunohistochemical estimation of their growth fraction and apoptotic status.

The proliferative and apoptotic status of undifferentiated carcinomas of the ovary was estimated immunohistochemically by antibody for Ki-67 proliferative antigen and Le(Y) apoptotic antigen. Well-differentiated types of ovarian carcinoma were examined for comparison. In 8 cases of undifferentiated carcinomas, the proliferative activity in terms of the growth fraction expressed as percentage of positive nuclei for Ki-67 antigen ranged from 28.2 to 81.2% (mean 60%). In well-differentiated carcinomas, which included 8 serous, 8 mucinous, 4 endometrioid and 4 clear cell adenocarcinomas, the growth fraction ranged from 8.2 to 33.4% (mean 20%). Thus, statistically significant differences were found in the proliferative activity between undifferentiated and well-differentiated ovarian carcinomas. On the other hand, apoptotic status as estimated by Le(Y) staining varied considerably among the cases in both undifferentiated and differentiated carcinoma groups. Nevertheless, in general, undifferentiated carcinomas seemed to be more prone to apoptosis than well-differentiated carcinomas. It is concluded that in ovarian undifferentiated carcinomas, more active cell proliferation takes place with a concomitant cell loss compared to their differentiated counterparts.

Expression of the proto-oncogene bcl-2 in uterine cervical squamous cell carcinoma: its relationship to clinical outcome.

We studied bcl-2 expression in patients with uterine cervical squamous cell carcinoma and correlated this phenomenon with survival. Immunohistochemical analysis with a monoclonal antibody specific for bcl-2 was used to detect the protein in tumor samples from 259 patients undergoing surgery for squamous cell carcinoma of the uterine cervix. Of the total, 67% (174) of the tumors were bcl-2 negative, and 33% (85) were positive. No significant difference in survival at five years was noted between patients with negative (78%) and positive (82%) tumors. However, when bcl-2 positive tumors were divided into partially stained (62 of 85, 73%), and diffusely stained (23 of 85, 27%) groups, the patients with partial staining had a better prognosis than those with diffused or negative (p < 0.01), staining (p < 0.05) (5 year survivals, respectively; 92%, 61%, and 78%). Though detection of bcl-2 positivity may itself not have clinical value for uterine cervical squamous cell carcinoma, the staining characters may add to predicting prognosis.

Basement membrane status in undifferentiated carcinomas of the ovary. Immunohistochemical distribution of type IV collagen and laminin.

Basement membrane status in well-differentiated and undifferentiated carcinomas of the ovary was examined and compared immunohistochemically by using antibodies which react with type IV collagen and laminin. In well-differentiated carcinomas, which included 8 serous, 8 mucinous, 4 endometrioid and 4 clear cell adenocarcinomas, the staining showed a regular and linear pattern in the interface between epithelium and stroma. However, in 8 cases of undifferentiated carcinoma, the staining for two antibodies was irregular, being either completely negative or focally positive in the portion surrounding the carcinoma cell nests. In some cases, intracytoplasmic staining was observed. Staining patterns for two antibodies were somewhat different within undifferentiated carcinoma cases. It is concluded that in ovarian undifferentiated carcinomas, the architecture of basement membrane is substantially altered compared to their differentiated counterparts.

Intramural adenocarcinoma of the uterus, arisen from adenomyosis uteri, showing unique histologic appearances. Report of two cases.

Two cases of uterine adenocarcinoma which grew mainly in myometrium and showed unique histologic appearances are reported. Both cases were considered to have arisen from adenomyosis uteri because transitional figures were observed between carcinoma cells and adenmyotic glands in one case, and carcinomatous glands were scattered in myometrium and were associated with endometrial stroma which mimicked benign adenomyosis in another. The former case was poorly differentiated adenocarcinoma and the latter was well-differentiated adenocarcinoma. Interestingly, in the latter case, the carcinoma cells were transformed into papillary adenocarcinoma such as has been observed in the thyroid gland, and formed well-demarcated nodular mass. These findings indicate that adenocarcinoma which arise from adenomyosis uteri could show various histologic appearances, in addition to usual endometrioid adenocarcinoma.