Evaluation of PTEN expression in cervical adenocarcinoma by tissue microarray

PTEN, a tumor suppressor gene, appears to negatively control the phosphoinositide 3-kinase signaling pathway for regulation of cell proliferation and cell survival. Somatic PTEN mutations are involved in a variety of tumors, including endometrial carcinomas, where PTEN expression is diminished. We examined expression of PTEN in a series of cervical adenocarcinomas and precursors, using tissue microarray (TMA) technology. TMA blocks were constructed using paraffin-embedded, formalin-fixed tissues from 273 samples derived from 16 normal cervical biopsies, 119 cases of invasive adenocarcinoma, and 20 high-grade cervical glandular intraepithelial neoplasia (CGIN). Fresh 3-mum sections were cut and immunostained with PTEN, and expression was correlated with clinicopathologic variables, including histologic subtypes of adenocarcinoma. In 137 patients, PTEN expression was positive in 121 (88%). The intensity and distribution of PTEN staining in the tumor tissue were more heterogeneous than those observed in the normal tissues. There were no significant differences in distribution or intensity of PTEN expression between adenocarcinoma in situ and subtypes of invasive adenocarcinoma. Our findings show that unlike the case in most endometrial carcinomas, PTEN expression is retained during the process of carcinogenesis in the glandular cervix. There is, however, evidence of altered distribution and intensity of PTEN expression in cervical adenocarcinoma cells.     

Cyclooxygenase-2 and c-erbB-2 expression in uterine cervical neoplasm assessed using tissue microarrays

OBJECTIVES: Cyclooxygenase-2 (COX-2) and c-erbB-2 are involved in the pathogenesis of solid organ tumors. Chemotherapeutic agents targeting COX-2 and c-erbB-2 are used to treat colon and breast cancers. This study evaluated the significance and relationship of COX-2 and c-erbB-2 protein expression in untreated uterine cervical neoplasm. METHODS: This study included 332 patients with uterine cervical neoplasm. We constructed tissue microarray blocks that included two cores from each donor tumor and immunostained them with primary anti-cyclooxygenase-2 and anti-c-erbB-2 monoclonal antibodies. The clinical features and survival data were compared. RESULTS: Three hundred and eighteen tumor samples (95.8%) could be interpreted after immunohistochemical staining. COX-2 protein expression was noted in 140 cases of uterine cervical neoplasm (44.0%): In 26.7% of the cervical intraepithelial neoplasia III (16/60 cases), 37.9% of the microinvasive squamous cell carcinoma (39/103 cases), 51.6% of the invasive squamous cell carcinoma (64/124 cases), and 76.2% of the adenocarcinomas (16/21 cases) (P < 0.005). By contrast, except for one case of adenoid cystic carcinoma, none of the uterine cervical neoplasm expressed c-erbB-2 protein. COX-2 protein expression correlated with histology (P < 0.005) and stage (P < 0.05), but was not associated with patient survival. CONCLUSION: COX-2 may participate in the progression of cervical squamous cell lesions, while the contribution of c-erbB-2 to cervical carcinogenesis is probably small.     

In situ adenocarcinoma and squamous carcinoma of uterine cervix. Pathological and immunohistochemical analysis with cytokeratin 13

OBJECTIVES: The aim of the study was the pathological and immunohistochemical analysis of cytokeratin 13 (CK13) in intraepithelial cervical tumors. STUDY DESIGN: We studied 415 in situ squamous carcinomas and 13 in situ mucinous cervical type adenocarcinomas of the uterine cervix. All patients underwent laser cervical conization and had a follow-up ranging 12-135 months. RESULTS: 3% of the squamous carcinoma patients recurred during the follow-up period, while the percentage of recurrence of in situ adenocarcinoma patients was 7.6%. We observed positive surgical edges in 46.1% of glandular tumors, and in 5% of squamous tumors. The percentage of recurrence was high among the cases with positive borders independently from their histopathologic type (14.3% in the squamous carcinomas versus 50% in the adenocarcinomas), compared to cases with negative edges (2.3% in the squamous carcinomas versus 0% in the adenocarcinomas). We observed CK13 positive staining in cervical squamous tumors and in mucinous cervical type adenocarcinomas, while there was no positive staining in non-neoplastic cervical glandular elements. CONCLUSION: CK13 positive immunostaining among in situ squamous and in situ mucinous cervical type adenocarcinoma cases adds additional evidence to data supporting a common origin of the two lesions.     

Tissue microarray: an effective high-throughput method to study the placenta for clinical and research purposes.

OBJECTIVE: Tissue microarray (TMA) technology allows simultaneous examination of the expression of many molecular markers (protein, mRNA, DNA, etc.) with high-throughput. The application of this technology, to date, has been largely confined to the study of cancer. Placental pathology poses unique challenges because of the size of the organ, its complex anatomy, as well as its histological heterogeneity. The objective of this study was to assess the feasibility and efficiency of TMAs for immunohistochemistry and in situ hybridization of placental tissues. STUDY DESIGN: TMAs were constructed using an automated tissue arrayer. Standard 0.6-mm or 1-mm microarray needles were used. Villous parenchyma, basal plate, and chorioamniotic membranes were targeted in each block. Five mum-thick TMA sections underwent immunohistochemical analysis of both cytoplasmic and nuclear antigens using a panel of antibodies against a variety of cytoplasmic [cytokeratin-7, vascular endothelial growth factor (VEGF), and protein Z], membranous (endoglin), and nuclear (c-fos and c-jun) antigens. mRNA in situ hybridization for surfactant protein A (SP-A) and chromogenic in situ hybridization for the Y chromosome (DYZ1) were also performed. RESULTS: Validation of TMA immunoreactivity demonstrated comparable results with corresponding whole sections. When a two-tiered scoring system (positive/negative) was employed, there was agreement between two and three cores and whole tissue sections (kappa>0.7). When a three-tiered scoring system (negative, weak-positive, or strong-positive) was used, the data from three cores showed the highest agreement with whole tissue sections (kappa >0.7). In situ hybridization experiments for mRNA and DNA were also successful in that the signals were readily detectable. Successful transfer from the donor block to the recipient block differed according to the anatomical compartment. The transfer efficiency of villous parenchyma, basal plate, and chorioamniotic membranes were 96.9% (875/903), 76.7% (115/150), and 75.4% (224/297), respectively. CONCLUSION: TMA is a practical and effective tool for high-throughput molecular analysis of the human placenta. Duplicate and triplicate cores offer agreement with whole tissue sections for two-category distinction immunostaining. TMA also affords relevant results from in situ hybridization experiments for mRNA and DNA. The major advantages are the conservation of tissues and reagents, simultaneous comparison of molecular markers in different anatomical compartments of the placenta, and reduction of experimental error.     

Tissue microarray: An effective high-throughput method to study the placenta for clinical and research purposes

Objective.?Tissue microarray (TMA) technology allows simultaneous examination of the expression of many molecular markers (protein, mRNA, DNA, etc.) with high-throughput. The application of this technology, to date, has been largely confined to the study of cancer. Placental pathology poses unique challenges because of the size of the organ, its complex anatomy, as well as its histological heterogeneity. The objective of this study was to assess the feasibility and efficiency of TMAs for immunohistochemistry and in situ hybridization of placental tissues.

Study design.?TMAs were constructed using an automated tissue arrayer. Standard 0.6-mm or 1-mm microarray needles were used. Villous parenchyma, basal plate, and chorioamniotic membranes were targeted in each block. Five μm-thick TMA sections underwent immunohistochemical analysis of both cytoplasmic and nuclear antigens using a panel of antibodies against a variety of cytoplasmic [cytokeratin-7, vascular endothelial growth factor (VEGF), and protein Z], membranous (endoglin), and nuclear (c-fos and c-jun) antigens. mRNA in situ hybridization for surfactant protein A (SP-A) and chromogenic in situ hybridization for the Y chromosome (DYZ1) were also performed.

Results.?Validation of TMA immunoreactivity demonstrated comparable results with corresponding whole sections. When a two-tiered scoring system (positive/negative) was employed, there was agreement between two and three cores and whole tissue sections (kappa>0.7). When a three-tiered scoring system (negative, weak-positive, or strong-positive) was used, the data from three cores showed the highest agreement with whole tissue sections (kappa >0.7). In situ hybridization experiments for mRNA and DNA were also successful in that the signals were readily detectable. Successful transfer from the donor block to the recipient block differed according to the anatomical compartment. The transfer efficiency of villous parenchyma, basal plate, and chorioamniotic membranes were 96.9% (875/903), 76.7% (115/150), and 75.4% (224/297), respectively.

Conclusion.?TMA is a practical and effective tool for high-throughput molecular analysis of the human placenta. Duplicate and triplicate cores offer agreement with whole tissue sections for two-category distinction immunostaining. TMA also affords relevant results from in situ hybridization experiments for mRNA and DNA. The major advantages are the conservation of tissues and reagents, simultaneous comparison of molecular markers in different anatomical compartments of the placenta, and reduction of experimental error.     

The histogenetic origin of cervical mesonephric hyperplasia and mesonephric adenocarcinoma of the uterine cervix studied with immunohistochemical methods.

Forty-four cases of mesonephric hyperplasia (MH) and two adenocarcinomas arising from mesonephric remnants (MA) in the cervix were compared immunohistochemically with 10 embryonic and fetal mesonephric tissues. The mesonephric cells retained their pattern for intermediate filaments during ontogenesis, as well as in the mature, hyperplastic, and neoplastic states: they expressed cytokeratin 8, cytokeratin 13, and vimentin, the two latter in variable amounts. In embryonic mesonephric tissues, cytokeratin was absent, whereas the staining for vimentin was intense. Fetal mesonephric cells stained for cytokeratin 13 and vimentin, but that staining diminished as maturation progressed. All MH and MA expressed cytokeratin 8, whereas only 20-30% of the cells in MH and 10-20% of carcinomatous mesonephric cells showed positive reactions with anti-cytokeratin 13 and anti-vimentin. CEA was always negative in cells of mesonephric origin. We regard these results to be important, since the reactions with anti-CEA and anti-vimentin enable one to distinguish cervical adenocarcinomas of mesonephric origin from those of endocervical origin, the latter being CEA-positive and vimentin-negative. Clinical studies revealed that approximately 75% of the patients with MH had used oral contraceptives for several years, 46% had precancerous lesions of the cervix, and in 62% the cervical mucosa showed adenomatous microglandular hyperplasia. We believe that hyperplasia of mesonephric remnants in the cervix may occur more often in patients with disturbed hormonal balance. However, the lack of a control population does not enable us to advance this hypothesis with assurance.     

A novel human monoclonal antibody against cervical cancer: its immunoreactivity with normal tube and ovary and with ovarian tumor tissue

1-1-2D, a novel human monoclonal antibody (MAb) raised against cervical cancer, was examined for its immunohistochemical reactivity with ovarian cancer. Six of 10 ovarian cancer cell lines showed positive staining, while 3 of 5 cervical cancer cell lines were positive. Among tumor tissues, 15 of 18 (83%) ovarian serous cystadenocarcinomas and 10 of 12 (83%) ovarian clear cell adenocarcinomas were positive. We also performed immunohistochemical staining of the same cancer specimens with OC 125 and compared their reactivity. The frequency of positivity was similar, but the reactivity of the two MAbs was different. 1-1-2D stained the apical surface of the glandular epithelial cells and secretory products of the gland. On the other hand, OC 125 stained the cytoplasm as well as the plasma membrane of the glandular epithelial cells. These results suggest that 1-1-2D MAb recognizes a different antigen from that recognized by OC 125.     

Immunohistological study of isoantigens, carcinoembryonic antigen and human chorionic gonadotropin in adenocarcinomas of the uterus

Isoantigens, carcinoembryonic antigen (CEA) and human chorionic gonadotropin (hCG) were simultaneously studied by immunohistology in the cervical and endometrial adenocarcinomas. Isoantigen loss was observed in all adenocarcinoma in situ (AIS) of the uterus. However, they appeared again in some tumor cells in 8 of 23 cervical and 4 of 41 endometrial adenocarcinomas. CEA was positive in 1 of 4 AIS and 22 of 23 adenocarcinomas of the cervix. It was distributed continuously in the apical portion and/or in the whole cytoplasm of tumor cells. On the other hand, CEA was positive only in 16 of 41 endometrial adenocarcinomas. It was strongly positive in the squamous elements, but weakly and sparsely in the apical portion and/or in the cytoplasm of some isolated glandular cells which occasionally showed a tendency to squamous differentiation. The majority of neoplastic glands, however, were negative for CEA. HCG was positive in 1 of 23 cervical and 3 of 41 endometrial adenocarcinomas. Two or 3 antigens were found concomitantly in some tumors, but localized differently from each other. It was concluded from the present study that the simultaneous examination of multiple antigens may be useful for characterization of adenocarcinomas of the uterus.     

The presence of HPV 16, 18 and p53 immunohistochemical staining in tumor tissue of Israeli Jewish women with cervical and vulvar neoplasia

The incidence of cervical neoplasia in Israeli Jewish women is persistently lower, while that of vulvar carcinoma is comparable to that in other populations. The aim of the present investigation was to assess the prevalence of HPV and of immunohistochemically detected mutant p53 in Israeli Jewish women with cervical and vulvar neoplasia compared with other populations. Tissue sections from formalin-fixed paraffin-embedded blocks of ten patients with CIN III, 29 with invasive squamous cell carcinoma, three with adenocarcinoma and 14 with invasive vulvar carcinoma, were examined for the presence of HPV 16 and HPV 18 DNA by PCR amplification, and for mutant p53 protein by immunohistochemical staining. HPV negative cases were re-examined with a sensitive primer. HPV DNA was detected in eight patients with CIN III and in 23 patients with invasive squamous carcinoma. In the remaining cervical squamous neoplasia tissue analysis with the sensitive primer could not be done. HPV DNA was also detected in two patients with adenocarcinoma and in nine (64.2%) patients with vulvar carcinoma. Positive p53 immunohistochemical staining was found only in one CIN III patient, in six (20.7%) squamous carcinoma and in 11 (78.6%) vulvar carcinoma patients. Of the p53 immunohistochemical staining positive tissues, two with cervical carcinoma and six with vulvar carcinoma were also HPV-positive. The prevalence of HPV and of positive p53 immunohistochemical staining in our series of Israeli Jewish women with cervical and vulvar neoplasia is similar to that in other populations, suggesting that the etiological factors are probably also alike.     

Gene expression profiling in two morphologically different uterine cervical carcinoma cell lines derived from a single donor using a human cancer cDNA array

PURPOSE: To examine the differentially expressed cancer-related genes in two morphologically different uterine cervical carcinoma cell lines derived from the same patient by an Affymetrix Human Cancer G110 Array carrying 1700 cancer-associated genes. In addition, to investigate specific gene expression depending on histological type, we examined expression of the selected genes in a panel of established cervical carcinoma cell lines derived from cervical adenocarcinoma and squamous cell carcinoma (SCC). EXPERIMENTAL DESIGN: Two distinct human uterine cervical carcinoma cell lines SKG-IIIa and SKG-IIIb derived from a single donor were screened using a cDNA microarray. The array results were additionally validated using semiquantitative RT-PCR. Expressions of the 10 selected genes were analyzed in the nine established cervical carcinoma cell lines using RT-PCR. RESULTS: The cDNA microarray analysis showed that 16 genes in SKG-IIIa were upregulated more than 10-fold compared to SKG-IIIb, and seven genes in SKG-IIIb were upregulated. Semiquantitative RT-PCR analysis of a subset of these differentially expressed genes gave results consistent with microarray findings. Among the 10 selected genes, insulin-like growth factor-binding protein-3, inhibitor of apoptosis protein 1, and cadherin-13 were more frequently expressed in SCC cell lines. 1-8D gene of interferon-inducible genes, Sno oncogenes, and transforming growth factor-beta II receptor were expressed in both SCC and adenocarcinoma cell lines. CONCLUSIONS: Our experimental data demonstrated that multiple genes are differentially expressed in uterine cervical carcinoma cell lines. It is suggested that these genes are involved with the differences in morphological characteristics and carcinogenesis of cervical carcinoma.     

Abnormal Expression of the Retinoblastoma Gene in Ovarian Neoplasms and Correlation to P53 and K-ras Mutations

We analyzed the expression of the retinoblastoma (Rb) gene in a group of ovarian neoplasms previously characterized for mutations in the p53 suppressor gene and the Ki-ras oncogene. Using immunohistochemical techniques, a total of 59 ovarian neoplasms spanning the histiologic spectrum from benign to malignant were examined for the expression of the Rb protein. All benign cystic adenomas and low malignant potential tumors exhibited normal expression of the Rb protein. Abnormalities in Rb protein staining were noted in 3 of 22 (14%) ovarian carcinomas. The staining patterns included tumors that were totally or focally negative for Rb protein. One tumor focally expressed Rb. This tumor demonstrated a direct juxtaposition of sections of Rb expressing and nonexpressing malignant epithelial cells. Two of the three tumors with abnormal Rb expression also had p53 mutations and staining on serial sections demonstrated that selected ovarian cancer cells possessed mutations in both oncogenes. These data suggest that the loss of Rb gene expression may play a role in the pathogenesis of a small number of invasive ovarian malignancies, but not in noninvasive ovarian neoplasms.     

Metastatic carcinoma of the gallbladder mimicking an advanced cervical carcinoma

BACKGROUND: Metastasis to the uterine cervix from a non-gynecologic neoplasm is extremely rare. To our knowledge, only three cases of primary carcinoma of the gallbladder with metastasis to the cervix have been previously reported. We report a case of metastatic gallbladder carcinoma mimicking a stage IIIB cervical carcinoma. CASE: A 74 year-old woman presented with pain in her left lumbar area radiating to her left flank. On physical examination, a 6 cm cervical tumor involving the left parametrium was noted. A computed tomography (CT) scan of the pelvis showed left-sided hydronephrosis and hydroureter with distal ureteral obstruction. A Pap smear revealed adenocarcinoma, and a biopsy of the endocervical canal was consistent with poorly differentiated adenocarcinoma. The patient was referred to a tertiary care center. On review of the biopsy after referral, the carcinoma was felt to be unusual for an endocervical primary and immunohistochemical stains were ordered. The carcinoma was positive for cytokeratin 7, had only focal cytoplasmic staining for p16, and was negative for carcinoembryonic antigen and cytokeratin 20. Upon presentation, the patient complained of new onset of shortness of breath and cough. A chest CT revealed multiple lesions in both lungs suggestive of metastatic disease, and an abdominal CT revealed a gallbladder tumor with extension into the liver. The patient underwent a CT-guided biopsy of one of the lung lesions and the pathologic findings were consistent with metastatic adenocarcinoma. The patient was diagnosed with stage IVB primary gallbladder adenocarcinoma and was treated with capecitabine, but her condition deteriorated rapidly and she died 5 months later. CONCLUSION: In patients with an atypical presentation for cervical adenocarcinoma, it is important to consider a metastatic tumor in the differential diagnosis and to perform a thorough work-up for metastatic disease before initiating therapy.     

Cytokeratin staining of resected lymph nodes may improve the sensitivity of surgical staging for endometrial cancer

OBJECTIVE: The presence of metastases to regional lymph nodes (LN) is the single most important risk factor in endometrial cancer. Advances in molecular biology have provided more sensitive methods for detecting micrometastasis. This was a pilot study to determine whether cytokeratin staining of LN from endometrial cancer patients is more sensitive than traditional histopathologic evaluation for the detection of micrometastasis. METHODS: The inclusion criteria included patients with surgical stage I-II endometrial cancer having >50% myometrial invasion, lesions >2 cm, and negative LN together with one of the following: FIGO grade 3 or cervical or lymph-vascular involvement. A matched control group included patients with LN metastasis. The evaluation of the LN at the time of initial surgery consisted of a frozen section and a reevaluation on permanent sections with H&E. In the study, lymphadenectomy specimens were cut, stained again with H&E and with cytokeratin, and examined. Cytokeratin staining was performed with AE1/AE3 antibodies. There were 16 LN-negative cases and 9 LN-positive controls. RESULTS: There was complete agreement between the LN assessment at time of surgery and the study H&E review prior to the staining for cytokeratin. However, 2 LN-negative cases (12.5%) had micrometastasis by cytokeratin staining. One of these patients developed recurrent disease in the para-aortic LN and died of disease at 2.8 years. CONCLUSION: Cytokeratin staining may improve the sensitivity for detection of metastasis compared to traditional evaluation. This study strongly suggests that these micrometastasis are clinically significant. An approach incorporating cytokeratin analysis could improve the risk assessment of specific patients.     

Umbilical metastasis (Sister Joseph''s nodule) as a first sign of a disseminated ovarian carcinoma: comparative immunohistochemical analysis of primary tumor and its metastases

The case of a 46-year-old female with umbilical metastasis as a first sign of an ovarian carcinoma is reported with the results of immunohistochemical analysis of primary tumor and lymph node and umbilical metastases. All specimens were positive for cytokeratin 7, CA 125, E-cadherin, alpha-, beta-, and gamma-catenin, as well as for MSH2. Staining with cytokeratin 20 and MLH1 was negative, and Ki-67 labeled from 5% (in the center of the lesions) to over 25% (at the periphery of the lesions) of the nuclei. Beta-catenin showed membranous positivity in the central parts and absence of staining at the periphery of ovarian tumor and umbilical metastasis, whereas lymph node metastasis presented with uniform reaction throughout. The results of immunohistochemical staining could point to the mechanisms employed by malignant tumors during invasion and growth of metastasis and suggest the possible role of the microenvironment in the expression of some adhesion molecules on tumor cells.     

高危型HPV DNA联合CK17检测早期宫颈鳞癌前哨淋巴结微转移的意义

目的检测早期宫颈鳞癌患者前哨淋巴结（sentinel lymph node,SLN）中人乳头瘤病毒（human papilloma virus,HPV）DNA和细胞角蛋白（CK）17的表达,探讨其在盆腔前哨淋巴结微转移中的临床意义。方法选取2012年1月至2013年1月山西医科大学第一医院早期宫颈鳞癌患者38例,其中ⅠB1期16例,ⅠB2期7例,ⅡA1期11例、ⅡA2期4例,实施经腹广泛性全子宫切除术＋盆腔淋巴结清扫术。术中采用亚甲蓝方法识别前哨淋巴结,对前哨淋巴结进行常规淋巴结病理检查、第二代捕获杂交（HC2）方法检测HPV DNA、免疫组织化学方法检测CK17。结果 38例早期宫颈鳞癌患者中SLN的检出率为86.84%,获得85枚SLN。在85枚SLN中淋巴结转移阳性率为21.18%（18/85）,高危型HPV DNA阳性率为34.12%（29/85）,CK17表达阳性率为30.59%（26/85）,二者均为阳性的有22枚。病理学检查为阳性的SLN,其高危型病毒检测和CK17检测均为阳性。病理组织学检查结果分别与高危型HPV DNA和CK17检测结果比较,差异有统计学意义（P〈0.05）。结论宫颈鳞癌淋巴结存在常规病理检测难以发现的微小转移,联合检测高危型HPV DNA和CK17的表达可提高SLN微转移检出率的准确性和特异性。     

Ancillary p16<Superscript>INK4a</Superscript> adds no meaningful value to the performance of ER/PR/Vim/CEA panel in distinguishing between primary endocervical and endometrial adenocarcinomas in a tissue microarray study

Purpose  Endocervical adenocarcinomas (ECA) and endometrial adenocarcinomas (EMA) are uterine malignancies that have differing biological behavior. The choice of appropriate therapeutic plan depends indeed on the tumor’s site of origin. In this study, we not only compare the individual expression status of five immunomarkers (ER, PR, Vim, CEA, and p16^INK4a), but also evaluate whether p16^INK4a adds value to the ER/PR/Vim/CEA panel characteristics in distinguishing between primary ECA and EMA. Methods  A tissue microarray (TMA) was constructed using paraffin-embedded, formalin-fixed tissues from 35 hysterectomy specimens, including 14 ECA and 21 EMA. TMA sections were immunostained with five anti-bodies, by avidin–biotin complex (ABC) method for antigen visualization. The staining intensity and area extent of the immunohistochemical (IHC) reactions were appraised by using the semi-quantitative scoring system. Results  The four respective markers (ER, PR, Vim, CEA) and their combined panel expressions showed significant (p < 0.05) frequency differences between ECA and EMA tumors. The p16^INK4a marker also revealed a significant frequency difference (p < 0.05) between the two sites of origin, but did not demonstrate to have any supplementary value to the 4-marker panel. Conclusion  According to our data, when there is histomorphological and clinical doubt as to the primary site of origin, we recommend that the conventional 4-marker (ER/PR/Vim/CEA) panel is appropriate. Ancillary p16^INK4a-marker testing does not add value to the 4-marker panel in distinguishing between primary ECA and EMA. C.-C. Yao and L.-F. Kok have equally contributed to this article.     

Immunohistochemical demonstration of carcinoembryonic antigen and ultrastructural features of endometrial adenocarcinoma

Determination of carcinoembryonic antigen levels in plasma (45 cases) and the immunohistochemical demonstration of tumor CEA (37 cases) were carried out in patients with endometrial adenocarcinoma. Twenty four out of 37 were also studied with the electron microscope. The plasma CEA level prior to therapy was significantly elevated (greater than 5.0ng/ml) in only one case (1/45:2.2%) of endometrial adenocarcinoma. CEA levels were more consistently elevated (4/17:23.5%) in patients with cervical adenocarcinoma. Immunoperoxidase staining of CEA (PAP method) was carried out using formalin-fixed paraffin embedded sections of 37 tumors. Tissue CEA activities were found in 9 out of 37 endometrial adenocarcinomas (24.3%) and 19 of 23 cervical adenocarcinomas (82.6%). Immunoreactive CEA was present in a high concentration on the cell surface of endometrial glands and less dense in their cytoplasm. In 9 of CEA positive endometrial carcinoma tissues, squamous metaplastic lesions were found in 4 cases and mucinous metaplasia in one case which were all CEA positive. With regard to the histopathological grade, CEA positive specimens were categorized G1 (4) and G2 (5) and all of the G3 were CEA negative. Seven of 9 cases of CEA positive specimens were examined under the electron microscope. There was no definite tendency in their ultrastructural characteristics, but all of the specimens examined revealed abundant cytoplasmic organelles suggestive of intracytoplasmic differentiation analogous to those of endometrial cells in proliferative phase. Moreover, fine structures of squamous and mucinous metaplastic cells were also described in detail.     

The cytokeratin profiles of ovarian common "epithelial" tumors

An improved immunohistochemical determination of the cytokeratin profiles of epithelia and their neoplasms is possible using monoclonal antibodies that will either identify all 19 cytokeratins (AE1/3) or delineate specific subsets (35 beta H11, 34 beta E12, 34 beta B4 and Cam 5.2). Ovarian common "epithelial" tumors (CET) contain cytokeratin filaments. To determine the nature and differences in the cytokeratin profiles of ovarian CET, eight benign Brenner tumors, four serous cystadenofibromas, 28 mucinous tumors, 27 serous tumors and six endometrioid, five clear cell and five undifferentiated carcinomas, as well as nine normal ovaries were immunostained with the above five antibodies. AE1/3 staining was predominant, while Cam 5.2 and 35 beta H11 displayed the most frequent staining thereafter. Statistically significant staining differences were found between a number of tumor groups using the antibodies 35 beta H11, 34 beta E12 and Cam 5.2. In this study, all ovarian CET, except the benign Brenner tumors, displayed a predominantly low molecular weight cytokeratin profile. The same profile in the normal surface epithelium lends credence to the belief that these tumors are derived from this epithelium. A significant staining difference between some of the tumor types using some of the antibodies suggests a possible ancillary, diagnostic role of cytokeratin profiling in situations where exact tumor typing is difficult.     

Immunohistochemical expression of molecular markers in an avian model: a potential model for preclinical evaluation of agents for ovarian cancer chemoprevention

OBJECTIVE: A significant obstacle confronting the evaluation of potential chemopreventive compounds in ovarian carcinoma is the absence of an animal model of spontaneous ovarian carcinogenesis. A potential model of adenocarcinoma has been described in the laying hen (Gallus domesticus). The purpose of this study was to evaluate the immunohistochemical expression of available antibodies that have been utilized in chemoprevention studies in this potential model of epithelial carcinoma. METHODS: Two hundred 2-year-old hens were sacrificed at Auburn University in accordance with IUACUC guidelines. Of these hens, 8 animals were thought grossly to have ovarian carcinoma and ascites. The tumors from these 8 hens were fixed in neutral-buffered formalin and processed to paraffin blocks. Hematoxylin and eosin stains were used to document the histologic presence of adenocarcinoma. Immunohistochemical evaluation for expression of antigen was performed using the following antibodies: CA125, CEA, cytokeratin, EGFR, erbB-2, Ki-67, Lewis Y, p27, PCNA, Tag 72, TGF-alpha, Muc 1, and Muc 2. RESULTS: Upon microscopic examination by a pathologist eight specimens were documented as adenocarcinomas. Several antibodies to antigens that are frequently expressed in human ovarian cancer were cross-reactive in the laying hen. Of these, cytokeratin AE1/AE3, pan cytokeratin, EGFR, Lewis Y, CEA, Tag 72, and erbB-2 stained the chicken carcinomas. EGFR and p185erbB-2 stained diffusely, and cytokeratin AE1/AE3, pan cytokeratin, Lewis Y, CEA, and Tag 72 were focally positive in the tumor. The aforementioned antibodies which have been useful as surrogate endpoints in chemoprevention trials and which also stained the chicken carcinomas included PCNA, p27, and TGF-alpha Antibodies that were not cross-reactive include CA 125, Ki-67, Muc 1, and Muc 2. CONCLUSION: The data presented in this pilot study support the potential utility of an avian model of spontaneously arising adenocarcinoma in which to study chemopreventive agents. More importantly, the influence of chemoprevention protocols on the expression of relevant antigens can be determined using available antibodies that are cross-reactive in this model. Thus, changes in the phenotypic expression of surrogate endpoint biomarkers as identified by cross-reactive antibodies can aid in the development of chemoprevention trials for human ovarian cancer.