Differential expression of WT1 and p53 in serous and endometrioid carcinomas of the endometrium.

Wilms' tumor antibody (WT1) has recently been reported to be reactive in most ovarian and peritoneal serous carcinomas, but few studies have looked at WT1 reactivity in endometrial carcinomas. p53, like WT1, is a tumor suppressor gene and in its mutated form is frequently present in endometrial serous carcinoma. Routine immunohistochemical staining for p53 and WT1 was performed in 70 endometrial carcinomas (39 endometrioid and 31 serous) of varying differentiation using tissue microarrays. Only 2 (7.5%) serous carcinomas and none of the endometrioid carcinomas (0%) were reactive for WT1. p53 immunoreactivity was found in 26 (83.9%) serous carcinomas and in 2 (5.1%) endometrioid carcinomas. We conclude that WT1 and p53 expression are not related and that WT1 expression in endometrial serous carcinoma differs from that of its extrauterine counterparts.     

基因芯片在筛选卵巢癌基因中的应用研究

目的:研究人卵巢癌相关基因在卵巢癌组织和正常卵巢组织中的差异表达。方法:用含512条癌及抑癌基因的cDNA表达谱芯片研究一组卵巢癌组织样本的基因表达谱。结果:共筛选出差异表达与癌相关的基因54条,其中18条表达上调,36条表达下调。结论:用基因表达谱芯片研究基因谱,可有效的筛选出新的卵巢癌相关基因。     

E-cadherin expression in endometrioid,papillary serous,and clear cell carcinoma of the endometrium

OBJECTIVE: To compare the frequency and pattern of E-cadherin expression in endometrioid, papillary serous, and clear cell carcinomas of the endometrium. METHODS: E-cadherin expression was examined in 76 endometrial carcinomas by immunohistochemistry using a monoclonal antibody to E-cadherin and was correlated with poor prognostic indicators such as depth of myometrial invasion, lymph node status, and intraperitoneal spread. The frequency of expression was compared between endometrioid, papillary serous, and clear cell carcinomas by the Fisher exact test. Logistic regression was used to examine the simultaneous effect of histological type and tumor grade on E-cadherin expression. RESULTS: Sixty-three endometrioid, nine papillary serous, two clear cell, and two carcinomas of mixed histology were examined. E-cadherin negative tumors were more likely to be poorly differentiated (P <.01), have cervical extension (P =.02), have positive peritoneal cytology (P <.01), and have adnexal spread (P =.01) when compared with E-cadherin positive tumors. Papillary serous and clear cell carcinomas were significantly less likely to express E-cadherin than endometrioid carcinoma (38% versus 95%, P <.001). Tumor grade and histological type were identified as significant predictors of E-cadherin expression in univariable analysis; however, only histological type remained significant in multivariable analysis (P =.01). When grade was controlled, endometrioid carcinoma remained 23 times more likely to express E-cadherin than papillary serous and clear cell carcinomas. CONCLUSION: Papillary serous and clear cell carcinomas are significantly less likely to express E-cadherin than endometrioid tumors. This difference may account for the more aggressive behavior of papillary serous and clear cell carcinomas.     

Apoptosis and the expression of Bcl-2 and Bax in patients with endometrioid, clear cell, and serous carcinomas of the uterine endometrium

OBJECTIVES: Clear cell and serous carcinoma of the uterus are rare types of endometrial carcinomas. This study was designed to investigate the differential occurrence of apoptosis, Bcl-2, and Bax in endometrioid, clear cell, and serous carcinomas. METHODS: In a total of 28 endometrial carcinomas as well as 4 samples of normal postmenopausal endometria, apoptotic changes were examined using molecular biochemical techniques. The expression of Bcl-2 and Bax proteins was also investigated by immunohistochemical staining with appropriate antibodies. RESULTS: Labeling of DNA in situ indicated that apoptotic cells were sporadically seen in postmenopausal endometrium (5.2 +/- 2.1, n = 4). In contrast, cells undergoing apoptosis apparently were detected in endometrioid carcinoma (29.3 +/- 3.7, n = 20), and their numbers increased intensely in clear cell (49.5 +/- 5.6, n = 5) and serous carcinomas (50.8 +/- 6.0, n = 3). Autoradiographic analysis revealed that high-molecular-weight DNA was predominant in postmenopausal endometrium. However, a DNA ladder was identified in 7 of 10 carcinomas. Although Bcl-2 was immunonegative or faintly immunopositive in all cases, many cases of endometrioid carcinoma (43.6 +/- 4.1%, n = 20) were immunopositive for Bax, unlike postmenopausal endometrium (17.6 +/- 6.7%, n = 4). Moreover, the number of cells expressing Bax increased in clear cell (60.4 +/- 6.5%, n = 5) and serous carcinomas (66.8 +/- 7.6%, n = 3) compared with that in endometrioid carcinoma. CONCLUSIONS: These results indicate that apoptosis occurs in a specific population of cells in different histologic components of endometrial carcinomas. The expression of Bax, but not of Bcl-2, might suggest histologic differentiation in endometrial carcinomas.     

Comparative immunohistochemical study of endometrioid and serous papillary carcinoma of endometrium.

OBJECTIVE: The aim of this study was to determine whether immunohistochemical analysis of molecular parameters can provide an alternative method for classification of endometrial cancer cases according to their aggressiveness. METHODS: Sixty-four cases of endometrial carcinoma were assigned to three groups: group I--28 cases of endometrioid well and moderately differentiated (G1-G2) carcinoma; group II--14 cases of endometrioid poorly differentiated (G3) carcinoma; group III--22 cases of serous papillary endometrial cancer. Immunohistochemistry was used to determine the existence of estrogen receptors (ER), progesterone receptors (PR), and the expression of bcl-2, p53, HER-2/neu and Ki-67. RESULTS: There was a significant difference in the immunohistochemical profile of the studied molecular parameters comparing the three study groups. The endometrioid G1-G2 cases (group I) were characterized by increased immunoreactivity for ER and PR (85.7% and 78.6%, respectively), increased immunoreactivity for bcl-2 (42.8%) and low expression of p53 (14.3%) and HER-2/neu (14.3%). In contrast to group I cases, the serous papillary endometrial cancer cases (group III) were characterized by immunonegativity for ER, PR and bcl-2 and high immunoreactivity for p53 (81.8%) and HER-2/neu (45.4%). The endometrioid G3 cases (group II) demonstrated an intermediate immunoprofile, characterized by immunonegativity for ER, PR and HER-2/neu, low immunoreactivity for bcl-2 (7.1%) and high expression of p53 (57.1%). The expression of Ki-67 did not differ significantly comparing the different cases of endometrial cancer. CONCLUSION: This study provides evidence that the immunohistochemical analysis of endometrial carcinoma differentiates between different grades and histological types, thus being useful in the distinction of high risk cases.     

Investigation of galectin-3 and heparanase in endometrioid and serous carcinomas of the endometrium and correlation with known predictors of survival

Introduction  

Inhibitors of tumor angiogenesis and metastasis are emerging as important new drug candidates for cancer therapy. Galectin-3 and heparanase have been shown to function in tumor progression and metastatic spread. Both of them exert pleiotropic effects; proliferation, cell migration, differentiation and tissue remodeling. The aim of this study was to investigate heparanase and galectin-3 expression in endometrioid and serous carcinomas of the endometrium and their relation with well-known prognostic factors, in addition to estrogen, progesterone, C-erbB-2, Ki-67 and p53.     

Opposite alterations of DNA methyltransferase gene expression in endometrioid and serous endometrial cancers

OBJECTIVE: To examine the DNA methyltransferase (DNMT) mRNA and protein levels in endometrioid and serous cancers and to study the relationship between DNA methyltransferase expression and endometrial cancer development. METHODS: Normal endometrium, Grade I and Grade III endometrioid carcinoma tissues and cell lines, as well as serous cancer tissues, were analyzed for DNMT expression. Real-time PCR and Western blot techniques were employed to measure the mRNA and protein levels of the four DNA methyltransferases, DNMT1, DNMT2, DNMT3A, and DNMT3B. Immunohistochemistry was performed to detect alterations in DNMT nuclear localization and spatial organization patterns. RESULTS: While DNMT2 and DNMT3A expression appear to be normal, two- to fourfold increase in DNMT1 and DNMT3B were found in both Grade I and Grade III endometrioid cancers. In addition, the poorly differentiated cell lines expressed relatively higher DNMT levels than well-differentiated cells. In contrast to endometrioid carcinomas, serous cancers expressed substantially lower levels of DNMT1 and DNMT3B than normal controls, with four- and twofold reduction observed in DNMT1 and DNMT3B mRNA levels, respectively. Western blot analysis confirmed opposite expression patterns of DNMT1 and DNMT3B protein in endometrioid and serous cancers. Immunohistochemistry showed normal nuclear localization of DNMT1 and DNMT3B in Type I and Type II cancer specimens as well as cell cultures. CONCLUSION: Two opposite DNMT expression patterns were identified in endometrioid and serous cancers. The concerted upregulation in maintenance and de novo DNA methyltransferases in endometrioid carcinomas is consistent with a tendency for gene-specific hypermethylation observed in this histologic subtype, and may be implicated in tumor suppressor silencing. In contrast, the downregulation of maintenance and de novo DNA methyltransferases in serous cancers suggests that these tumors may contain hypomethylated genomic DNA, which has been associated with a higher mutation rate and is consistent with the known pathogenesis of serous-specific phenotypes. Taken together, the data suggest that divergent DNA methylation pathways may be implicated in the development of Type I and Type II endometrial cancers.     

Wilms tumor gene immunoreactivity in primary serous carcinomas of the fallopian tube, ovary, endometrium, and peritoneum.

Wilms tumor gene (WT-1) expression has been reported in many human cancers, including most ovarian and peritoneal serous carcinomas, but has not been studied in carcinomas of the fallopian tube. In this study, the authors evaluated the immunohistochemical expression of WT-1 in serous carcinomas of the fallopian tube and compared their reactivity with that of ovarian, peritoneal, and endometrial serous carcinomas. All primary serous carcinomas of the fallopian tube (13 cases), ovaries (25 cases), and peritoneum (3 cases) were reactive with the WT-1 antibody, whereas all five primary endometrial serous carcinomas were nonreactive. WT-1 reactivity in an unknown primary serous carcinoma is therefore suggestive of an extrauterine site. The marked difference in WT-1 staining raises the possibility of genetic differences between serous carcinomas arising in the endometrium compared with those arising in the ovaries, fallopian tubes, and peritoneum.     

WT1 is differentially expressed in serous, endometrioid, clear cell, and mucinous carcinomas of the peritoneum, fallopian tube, ovary, and endometrium.

The Wilms' tumor gene WT1 plays complex roles in the development of the organs of the genitourinary tract and mesothelium, as well as Wilms' tumors. Although its biologic role is still unclear, most serous carcinomas of the ovary and peritoneum, mesotheliomas, and Wilms' tumor have been shown to express WT1. A recent study, however, found no WT1 expression in serous carcinomas of the endometrium, suggesting that WT1 could be useful in identifying the primary site of serous carcinomas. We examined the expression of WT1 and p53 by immunohistochemistry in 69 cases of endometrial carcinoma (35 endometrioid, 18 clear cell, 16 serous), 68 cases of ovarian carcinoma (28 serous, 11 endometrioid, 18 clear cell, and 11 mucinous), 14 fallopian tube carcinomas (12 serous, 2 endometrioid), and 20 primary peritoneal serous carcinomas. WT1 nuclear reactivity of any extent and intensity was considered positive. Immunohistochemical stains were evaluated semiquantitatively using a four-tiered scale. Among endometrial carcinomas, WT1 immunoreactivity was seen in 10 of 16 serous, but in none of 35 endometrioid or 18 clear cell carcinomas. Among ovarian tumors, WT1 expression was seen in 24 of 28 serous and 4 of 18 clear cell carcinomas, but in none of 11 endometrioid and 11 mucinous tumors. All 12 serous carcinomas but none of 2 endometrioid carcinomas of the fallopian tube were positive for WT1. WT1 expression was seen in 19 of 20 serous primary peritoneal carcinomas. The difference in WT1 expression was highly significant between serous and other types of tumors in all sites (p<0.0001, chi-square test), although the level of WT1 expression was significantly different among serous carcinomas arising at different sites (p<0.0001, Kruskal-Wallis test). A significant positive correlation was found between the level of p53 and WT1 expression in all carcinomas combined (r = 0.3935, p<0.0001, Spearman test), but when only serous carcinomas were analyzed, the correlation between p53 and WT1 expression levels did not reach statistical significance. Our results suggest that WT1 expression in epithelial tumors of the female genital tract may be related to cell differentiation and histologic subtypes of carcinomas, rather than to primary site of origin.     

Outcome of uterine clear cell carcinomas compared to endometrioid carcinomas and poorly-differentiated endometrioid carcinomas

OBJECTIVES: Our aim was to compare the survival between patients with clear cell carcinoma (CC) and patients with endometrioid carcinoma (EC). METHODS: Through the population-based Geneva Cancer Registry, we identified 1,380 resident women diagnosed with uterine cancer between 1970 and 2000. We excluded those with papillary serous endometrial carcinoma and uterine sarcomas. We categorized patients as CC (n = 32, 2.8%) or EC (n = 1,145, 97.2%). Uterine cancer-specific survival rates were calculated by Kaplan-Meier analysis. We used Cox proportional hazards analysis to compare uterine cancer mortality risks between groups, and adjusted these risks for other prognostic factors. RESULTS: CC patients presented with a more advanced stage at diagnosis than EC patients (p = 0.002). Compared to women with EC, women with CC had a significantly greater risk of dying from their disease (hazard ratio [HR] 2.9, 95% confidence interval (95% CI) 1.7-4.9). After adjustment for age, stage and adjuvant chemotherapy, the risk of dying from uterine cancer was still significantly higher for CC patients (HR 2.0, 95% CI 1.2-3.4). By univariate analysis, the risk of dying of endometrial cancer was not significantly higher in CC patients than in patients with poorly-differentiated EC (HR 1.3, 95% CI 0.7-2.3). CONCLUSION: This population-based investigation shows that patients with CC have a poorer outcome than those with EC. Studies to determine the role of adjuvant treatment in CC patients are needed.     

Benign and malignant serous and endometrioid epithelium in the omentum.

OBJECTIVES: Benign and malignant serous and endometrioid epithelial proliferations are found in the omentum, where their presence may be interpreted either as metastases from Müllerian tumors elsewhere or as primary peritoneal tumors. The present study was undertaken in an attempt to gather data that might help resolve the issue. METHODS: The ratios of serous epithelium to endometrioid epithelium in the omentum, in both the benign and malignant states, were determined for cases from January 1985 to July 1997 and January 1991 to December 1997, respectively. RESULTS: In ovarian carcinoma, the ratio of malignant serous epithelium to endometrioid epithelium involving the omentum is 15:1. This is comparable to the ratio of benign serous epithelium to endometrioid epithelium in the omentum, which is 10:1. The ratio of primary peritoneal serous carcinoma to endometrioid carcinoma is 10.5:1. CONCLUSION: It seems not reasonable that endometrioid carcinoma of the ovary is 15 times less likely to metastasize to the omentum than its serous counterpart. The ratio, however, is not unreasonable if endometrioid and serous carcinomas arise from preexisting endometrioid or serous epithelium. We conclude that serous and endometrioid carcinomas may arise primarily in the omentum and, in at least some cases, may derive from their benign counterparts.     

Combined large cell neuroendocrine and endometrioid carcinoma of the endometrium.

We present the surgical and pathological findings and follow-up of 5 women diagnosed with combined endometrioid and high-grade neuroendocrine carcinoma of large cell type (LCNEC) arising in the endometrium. The mean age of the women was 75 years (range, 50-88 years). Of the 5 tumors, 4 formed polypoid endometrial masses associated with extensive lymphovascular involvement of the myometrium by neoplastic cells. A single endometrial tumor was formed by LCNEC alone, and 4 tumors were composite with varying proportions formed by endometrioid (4/5) and small cell neuroendocrine carcinoma (1/5). In all 5 LCNEC tumor components, an insular growth pattern was noted, whereas a diffuse (solid) pattern was found in 4 tumors, a trabecular in 2, and rosettes/pseudorosettes in another 2. In all 5 tumors, the LCNEC tumor components were labeled with neuron-specific enolase (NSE). Four tumors were reactive for chromogranin A, CAM 5.2, and p53. Three tumors were labeled for AE1/AE3, CD56 (NCAM), p16, and cytokeratin 7. Synaptophysin was reactive in 2 tumors, and CD117 was found in only a single tumor. Of the 3 endometrioid tumor components examined, all were reactive for NSE. Two tumors were reactive for p16 and p53, 1 for CD56, but none for synaptophysin orchromogranin A. We conclude that LCNEC of the endometrium is a distinct clinicopathological entity with a poor prognosis irrespective of stage. The gross and histomorphological features are often suggestive, but confirmation requires immunoperoxidases, including NSE, synaptophysin, chromogranin A, p16, and p53. Combined endometrioid and high-grade LCNEC possess more characteristics of a type II than a type I endometrial carcinoma.     

Neutral mucosubstances as a prognostic indicator in serous and endometrioid carcinomas of the ovary. A morphometrical analysis of saliva-periodic acid-Schiff stain

Forty-four serous and 17 endometrioid carcinomas of the ovary were classified on hematoxylin-eosin stained tissue sections using the criteria of World Health Organization (WHO). The amount of saliva-periodic acid-Schiff (SPAS) staining positivity, indicating presence of neutral mucosubstances, was quantitated morphometrically and compared with survival of the patients. The 5-year survival of patients who had SPAS positive serous carcinoma was 36% whereas that of patients with SPAS negative serous carcinomas was only 7%. Among the patients who had endometrioid carcinoma showing SPAS positivity over the median, the 5-year survival was 47%. All patients with SPAS positivity less than the median had died in 3 years after the diagnosis. The differences in survivals were statistically significant (P < 0.001). This study shows that neutral mucosubstances as quantitated morphometrically is a good prognostic indicator of serous and endometrioid carcinomas of the ovary.     

Analysis of p53 and c-erbB-2 expression in ovarian endometrioid carcinomas arising in endometriosis.

We assessed the immunohistochemical expression of p53 and c-erbB-2 oncoproteins in 13 ovarian endometrioid adenocarcinomas arising from endometriosis (group 1) and compared the findings with 15 otherwise similar cases without associated endometriosis (group 2). Tumors in group 1 showed a higher expression of both p53 and c-erbB-2 (p = 0.015 and p = 0.048, respectively). The expression of the two proteins was also significantly associated in group 1 (p = 0.013) but not in group 2 (p = 0.63) tumors. The different pattern of expression of p53 and c-erbB-2 in the two groups suggests that different molecular pathways may be involved in their pathogenesis.     

Pelvic and para-aortic lymphadenectomy in patients with endometrioid adenocarcinoma of the endometrium.

BACKGROUND: The purpose is to determine the rate of lymph node metastases in women with endometrioid adenocarcinoma of the endometrium (EAE) undergoing systematic lymphadenectomy. METHODS: Patients (349) underwent a complete pelvic and para-aortic lymphadenectomy from caudal to the median circumflex to the level of the renal vessels. RESULTS: Grade 1 tumors accounted for 32.7% of the tumors and 31.0% of the positive nodes, grade 2 accounted for 47.3% of the tumors (37.9% of positive nodes), and grade 3 accounted for 20.1% of the tumors and 31.0% of the positive nodes (P>0.05). Positive nodes were found in 15.8% of grade 1 tumors, 13.3% of grade 2 tumors and 25.7% of grade 3 tumors (P>0.05). Isolated para-aortic involvement without pelvic nodal involvement occurred in 29% of patients with positive nodes. CONCLUSIONS: When complete lymphadenectomies are performed in EAE, positive lymph nodes (including isolated para-aortic lymph nodes) are common in all grades.     

Molecular classification of human endometrial cancer based on gene expression profiles from specialized microarrays

Objective

To investigate whether the molecular classification of endometrial cancer based on gene expression profiles can predict the biological behavior of the tumors and inform prognosis.

Methods

An array containing 492 genes was used to generate gene expression profiles from 35 tumor samples. A hierarchical cluster algorithm was used to compare gene expression patterns among the tumor samples.

Results

A cluster analysis revealed 3 distinct tumor clusters. A comparative analysis of tumor type, grade, FIGO stage, and depth of myometrial invasion revealed significant differences in grade and stage among the clusters, which appear to group tumors with specific clinical behaviors. Moreover, the cluster analysis initially revealed 2 clusters of differentially expressed genes. One contained 38 genes that were upregulated in most samples of the cluster representing the most advanced disease, and the other contained 27 genes that were upregulated in most samples of the cluster representing the least advanced disease.

Conclusion

Molecular classification of endometrial cancer based on gene expression profiles obtained by designing specialized microarrays indicated a marked correspondence with the histologic features and clinical behavior of endometrial cancer tumors.     

Progesterone-regulated gene expression in the primate endometrium

Progesterone action is essential for maturation of the endometrium to a receptive state for implantation in humans and nonhuman primates. The orchestration of progesterone-regulated gene expression is also temporally controlled during the secretory phase based on the limited window for implantation. The genes and gene networks affected by progesterone are likely to involve both activation and repression. Our laboratory has used the rhesus monkey as a model to study the regulation of genes known or suspected to be involved in endometrial maturation. In addition, we have used subtractive hybridization and differential display techniques to identify novel or unsuspected genes that are regulated by progesterone during endometrial maturation. Our studies have led us to propose a working model of progesterone action during the primate secretory phase that includes waves of gene activation and repression that culminate in a receptive endometrium.