High incidence of breast and endometrial neoplasia resembling human Cowden syndrome in pten+/- mice.

PTEN is one of the most commonly mutated tumor suppressor genes in human cancer. PTEN mutations have been implicated in the development of a variety of human neoplasia, including high-grade glioblastoma, prostate, breast, endometrial, and thyroid carcinoma. Germ-line mutations of PTEN cause Cowden's syndrome (CS), a multiple hamartoma condition resulting in increased susceptibility for the development of cancer. When more than 6 months old, pten+/- mice develop a range of tumors, partially resembling the spectrum of neoplasia observed in CS patients. One-half (32 of 65) of pten+/- females developed breast tumors, whereas all (65 of 65) of the females had endometrial hyperplasia, and there was a high incidence (14 of 65) of endometrial cancer. Hamartoamous tumors of the gastrointestinal tract, as well as prostate and adrenal neoplasia, were also frequently observed. Significantly, the spectrum of neoplasia observed in pten+/- mice partially overlaps with the types of tumors frequently detected in CS patients. The majority of tumors in pten+/- mice exhibit loss of heterozygosity at the pten locus, which indicates the importance for loss of PTEN function in tumor formation. Consistent with the role of PTEN in negative regulation of PKB/Akt phosphorylation and activity, pten loss of heterozygosity is accompanied by hyperphosphorylation of PKB/Akt in tumors. Taken together, our results establish pten+/- mice as an excellent animal model system for the investigation of PTEN-related hamartoma syndromes, as well as the role of PTEN in breast and endometrial carcinogenesis.

Role of a DT-diaphorase mutation in the response of anal canal carcinoma to radiation, 5-fluorouracil, and mitomycin C.

PURPOSE: To determine, retrospectively, the status of the bp 609 mutation in the DT-diaphorase gene in anal canal carcinoma patients who have undergone radical radiotherapy with concurrent 5-fluorouracil (5-FU) and mitomycin C (MMC), to determine the relationship of the mutant form of the gene to treatment outcomes. METHODS AND MATERIALS: Paraffin blocks of pretreatment tumor biopsies were obtained on 49 patients who underwent treatment with curative intent using radiation, infusional 5-FU and bolus MMC from January 1991 to December 1993. DNA was extracted and subjected to polymerase chain reaction (PCR) analysis using primers that encompassed the bp 609 C to T mutation. Restriction endonuclease cleavage with Hinf 1 and gel electrophoresis were used to determine the polymorphism status of each patient. RESULTS: DNA of 46 patients was successfully amplified. The 46 patients were distributed as follows: 26 (56.5%) C/C-homozygous wildtype, 18 (39%) T/C-heterozygous, and 2 (4.5%) T/T-homozygous mutant. Eleven of 46 patients had suffered treatment failure. The status of the bp 609 polymorphism in this group was 5 (45.5%) C/C, 5 (45.5%) C/T, and 1 (9%) T/T. CONCLUSION: In this series, there was not an overrepresentation of the mutant allele in patients with treatment failure, suggesting that the bp 609 alteration is not a strong determinant of treatment outcome.

Prognostic role of p53 protein expression in epidermoid carcinoma of the anal canal.

PURPOSE: To assess the prognostic significance of p53 protein expression in patients with primary epidermoid carcinoma of the anal canal managed by radiation therapy (XRT), 5-fluorouracil (5-FU), and mitomycin C (MMC). METHODS AND MATERIALS: From January 1991 to December 1993, 58 consecutive patients with primary epidermoid carcinoma of the anal canal were treated in a prospectively designed protocol of XRT (24 Gy/12--3(1/2) wk split--28 Gy/14) and concurrent 5-FU (1000 mg/m2/day 1-4) and MMC (10 mg/m2 day 1) of each cycle of XRT. Paraffin-embedded tumor samples were unavailable in 9 patients, leaving 49 patients in the study. Expression of p53 protein was studied using immunohistochemistry and quantified as percent tumor nuclei showing positive staining. Actuarial survival and disease-free survival (DFS) rates were estimated by the Kaplan-Meier method, and compared using the log-rank test. A Cox proportional hazard model was used for the multivariable analysis. RESULTS: There were 6 T1, 26 T2, 7 T3, and 10 T4 lesions. Primary tumor sizes ranged from 1-15 cm with a median of 4 cm. There were 6 patients with nodal metastases. Median follow-up was 4.5 years. Positive nuclear immunostaining for p53 was observed in 40 of 49 patients. The median percent positive staining was 5%, with 13, 9, and 18 patients showing staining in <5%, 5 to <10%, and 10-50% of tumor nuclei respectively. There was no correlation of percent p53 staining with gender, age, tumor stage, size, or histology. Local, regional, and distant failures were observed in 12, 2, and 2 patients respectively. The 5-yr survival and DFS were 84% and 64% respectively. In univariate analysis, the only prognostic variable for survival was gender. For DFS, advanced T category and large tumor size were predictive of poor DFS. In multivariate analysis, poor DFS was associated with high T category (p = 0.0008), basaloid histology (p = 0.001), male gender (p = 0.002), and increasing percent of p53 protein expression (p = 0.01). CONCLUSIONS: It is concluded that expression for p53 protein is present in a high percentage of patients with epidermoid carcinoma of the anal canal. For patients managed with combined XRT, 5-FU, and MMC, percent p53 protein expression is of prognostic value for DFS independent of other clinical factors such as T category, gender, and histology.

Expression of the verotoxin receptor glycolipid, globotriaosylceramide, in ovarian hyperplasias.

The presence of cell surface receptor glycolipid, globotriaosylceramide (Gb3), is essential to confer susceptibility to the E. coli-derived verotoxin (VT). Our earlier studies showed that Gb3 is expressed in ovarian carcinoma cell lines. The Gb3 content of normal ovary, benign and malignant primary ovarian tumors, and their metastases have now been compared by verotoxin thin-layer chromatogram (TLC) overlay of the glycolipid tissue extracts. FITC-labeled VT1 B subunit binding to frozen tumor sections was also monitored histochemically. Low to undetectable levels of Gb3 were found in "normal" ovarian tissue. Gb3 was markedly increased in both benign and malignant tumors, suggesting that increased Gb3 may be related to proliferation, rather than malignancy per se. Mucinous tumors showed the least Gb3 elevation; serous tumors were variable, showing higher levels of Gb3 in less differentiated malignant tumors. By far the highest Gb3 content was observed for secondary ovarian metastases and tumors refractory to chemotherapy. Frozen sections of neoplastic ovarian tissue overlaid with fluorescein-conjugated VT1 B subunit show extensive binding to tumor cells, particularly in poorly differentiated samples and blood vessels adjacent to, and within, the tumor mass. Tumor foci were stained but stromal tissue was consistently negative both in primary tumors and metastases. VT staining of well-differentiated primary ovarian tumor sections was weak, corresponding to their low Gb3 content, but strong staining was observed in sections from a highly differentiated primary tumor from a patient who was unexpectedly refractory to clinical chemotherapy. These studies suggest that verotoxin/Gb3 targeting may provide the basis for new treatments for ovarian cancer.

Colposcopy of intraepithelial neoplasia of the vulva and adjacent sites.

The colposcopic appearances of VIN III differ on the vulva and adjacent sites according to the anatomic sites. Lesions on the hairy skin appear more well defined than lesions on nonhairy areas. VIN III histologically can extend into underlying adnexal structures. Most frequently, it is the hair follicles in which it may extend deeply. This involvement is not identified by colposcopic methods. A variety of colposcopic mimics or "look-alike" lesions occur on the vulvar skin. Therefore the performance of a biopsy is required to establish the diagnosis.

Developments in the pathology of ovarian tumours.

The understanding of the pathogenesis and pathology of ovarian tumours is constantly evolving. Dominant themes in recent studies of ovarian tumours include prognostic features in borderline tumours, molecular events in the pathogenesis of ovarian tumours, the assessment of tumour features that may have prognostic or treatment implications, and the development of techniques that may enhance diagnostic accuracy. The literature has been reviewed with an aim to identifying those studies that can potentially impact practice and improve patient care. The most noteworthy developments include: the understanding that so-called 'tumours of low malignant potential' are virtually always benign, and that one can identify those rare cases with malignant potential; the importance of the recognition of micropapillary serous carcinomas; an improved understanding of early invasive carcinomas and their impact on screening practices; an understanding of the association of endometriosis with ovarian cancer; further awareness of factors in ovarian tumours that influence prognosis, such as refinements in grading and molecular markers such as P27; and refinements in diagnosis so as to distinguish primary from metastatic cancer and benign lesions from malignant tumours more effectively.

Female adnexal tumor of probable wolffian origin: a clinicopathological case report and a possible new treatment.

Female adnexal tumors of probable wolffian origin (FATWOs) are rare tumors arising in the broad ligament from the remnants of the mesonephric duct. We report a case of recurrent disease. A 15-year-old girl who presented with a painful pelvic mass underwent a laparotomy with tumor resection. Pathology findings confirmed a FATWO. The tumor recurred within 2 years and was treated with multiple chemotherapy regimens, including a platinum-based drug, and surgery for progressive disease. The tumor was positive for c-kit oncogene (CD 117). Gleevac therapy, a tyrosine kinase inhibitor, was prescribed, and she developed severe persistent lower abdominal pain 2 months later. She underwent a hysterectomy and debulking of retroperitoneal masses. Pathology showed evidence of tumor necrosis, suggesting a possible beneficial effect, and she was recommenced on Gleevac in an effort to prevent recurrences. She is currently asymptomatic, without evidence of disease 10 months after surgery, continuing on Gleevac therapy. FATWOs are very rare tumors. Most cases are benign but have the potential to recur and metastasize. There is limited knowledge about the optimal treatment for this neoplasm. Our patient's favorable response to Gleevac therapy supports the concept of targeted molecular therapy in patients with c-kit-positive FATWO tumors.

MSN-1 antibody in the evaluation of female genital tract adenocarcinomas.

The reactivity of a new monoclonal antibody (MAb), MSN-1, raised against a human endometrial cancer cell line (SNG-II), was studied in a variety of endometrial, endocervical, and ovarian carcinomas as well as normal cycling endometrium. Moderate to strong reactivity (2-3+) was seen in six of nine normal secretory endometria (67%), one of 10 normal proliferative endometria (10%), 18 of 18 endometrial adenocarcinomas (100%), 10 of 11 endometrioid ovarian adenocarcinomas (91%), seven of nine clear cell ovarian adenocarcinomas (78%), one of 12 endometrial hyperplasias without atypia (9%), two of four endometrial hyperplasias with atypia (50%), zero of five endometrial serous adenocarcinomas, two of 17 serous ovarian adenocarcinomas (12%), zero of 10 intestinal-type mucinous ovarian adenocarcinomas, and zero of nine metastatic adenocarcinomas in ovary. Endocervical adenocarcinomas showed moderate to strong staining in 75% (six of eight). It is concluded that MSN-1 can be used to confirm endometrioid/clear cell differentiation in ovarian and endometrial tumors, cannot be used to discriminate endocervical from endometrial differentiation, cannot be used to discriminate atypical hyperplasia from carcinoma, and may be useful to distinguish between atypical (premalignant) endometrial hyperplasias and those without atypia.

Adenomyosis: US features with histologic correlation in an in-vitro study.

PURPOSE: To evaluate the accuracy of ultrasonographic (US) features of adenomyosis by correlating them with histologic findings and to assess inter- and intraobserver agreement. MATERIALS AND METHODS: US was performed and videotaped in 102 consecutive hysterectomy specimens in a water bath. Videotapes were reviewed initially by two independent radiologists blinded to the clinical and histologic findings and after 1 month by one of the two; US and histologic findings were correlated. Features evaluated included diffuse abnormal echotexture of myometrium, subendometrial myometrial cysts, subendometrial echogenic nodules, subendometrial echogenic linear striations, nodular endometrial-myometrial junction, poor definition of the endometrial-myometrial junction, asymmetric thickness of the anteroposterior wall of the myometrium, and globular configuration. RESULTS: The prevalence of adenomyosis in this cohort was 29.4% (30 of 102 specimens). The mean sensitivity, specificity, negative predictive value, positive predictive value (PPV), and accuracy for the diagnosis of adenomyosis for the three reviews were 81%, 71%, 90%, 54%, and 74%, respectively. All findings evaluated, except for nodular endometrial-myometrial junction, were significantly more common in uteri with adenomyosis (P <.05). Heterogeneous myometrium reached borderline significance (P =.05). The specificities and PPVs of subendometrial striations, subendometrial echogenic nodules, and asymmetric myometrial thickness were significantly higher than those of other features (P <.05). The interobserver agreement was moderate (kappa = 0.48), and the intraobserver agreement was good (kappa = 0. 67) for the three reviews. CONCLUSION: The presence of subendometrial linear striations, subendometrial echogenic nodules, or asymmetric myometrial thickness improves the specificity and PPV of US in diagnosing adenomyosis.

Uterine sarcoma: an analysis of 74 cases.

In order to determine whether recent methods of diagnosis and treatment have resulted in an improved survival for patients with uterine sarcoma, we reviewed 99 cases treated in our center from 1970-1985. Morphologic characteristics of 74 tumors were specifically reassessed for this study. All tumors were graded. Of 42 Stage I cases that were morphologically assessed, tumor-positive pelvic lymph nodes were found in two of the 15 patients in whom sampling was done. No cases of tumor-positive para-aortic nodes were found in 14 patients with Stage I disease. In Stage I and Stage II, no cases of positive para-aortic nodes were found in association with negative pelvic nodes. The 2- and 5-year survival rates in Stage I were 47.4% and 29.4%, respectively. Local recurrence decreased (p less than 0.01) in Stage I from nine of 22 cases in which operation alone was performed to none of 15 cases in which pelvic radiotherapy was added, but no improvement in the 5-year survival rate was observed. As with lymphadenectomy and radiotherapy, the recent use of chemotherapy for uterine sarcoma had no impact on survival.

Epidermal growth factor receptor expression and the presence of human papillomavirus in cervical squamous intraepithelial lesions.

To determine the relationship between the expression of epidermal growth factor receptor (EGFR) and the presence or type of human papillomavirus (HPV) in cervical squamous intraepithelial lesions (SIL), paired colposcopically directed cervical biopsies were obtained from 88 patients referred for abnormal Papanicolaou smears. One biopsy was formalin-fixed and processed for conventional light microscopy, and the other was immediately frozen. A portion of the frozen tissue was used for Southern blot HPV DNA hybridization and a portion for immunohistochemical studies for EGFR using a monoclonal antibody. Forty-seven cases were SIL and 41 were normal. In 41 (87%) of the cases of SIL and in eight (20%) of the normal cases, HPV DNA was detected. Of the SIL cases, HPV 16 was the most frequently detected type, being present in 12 (25%), followed by 10 (21%) types 31 or 35, nine (19%) types 52 or 56, five (11%) uncharacterized types, three (6%) type 18, and two (4%) multiple types. Regardless of histology, EGFR was detected in all cases. In normal cases, EGFR expression was detected in the basal cells only and in SIL in abnormal proliferating parabasal cells such that it correlated with the grade of SIL. When stratified by grade of SIL, no differential expression of EGFR was seen in cases where HPV was detected; in contrast, in cases where no HPV was detected, no differential expression was seen between cases of different HPV type. Thus, EGFR is expressed by all proliferating squamous epithelial cells and as such correlates with the grade of SIL.(ABSTRACT TRUNCATED AT 250 WORDS)

Primary localized amyloidosis of the lacrimal glands.

Primary localized amyloidosis causing bilateral lacrimal enlargement is rare. The pathogenesis of amyloid deposition within the orbit and other body tissues has not been fully elucidated. The authors report the case of a 72-year-old woman who presented with bilateral lacrimal gland enlargement secondary to amyloid infiltration. The chemical nature of the deposit was characterized using light microscopy, immunohistochemistry, and immunoelectron microscopy. The primary (immunocytic) nature of the amyloid was confirmed by immunohistochemistry demonstrating the presence of monoclonal lambda light chains in the amyloid deposits and in the plasma cells. Using immunoelectron microscopy, amyloid deposits were seen containing lambda light chains in macrophages. It has been postulated that the macrophage has a role in amyloid deposition. The authors believe this to be the first published report of immunoelectron microscopy use in orbital amyloidosis, and that this technique has helped further their understanding of the nature and pathogenesis of this condition.

Evaluation of two commercially available in situ hybridization kits for detection of human papillomavirus DNA in cervical biopsies: comparison to Southern blot hybridization.

In order to determine the distribution of human papillomavirus (HPV) types and to compare in situ hybridization (ISH) with Southern blot hybridization (SBH), paired colposcopically directed cervical biopsies and cervical-vaginal lavages were obtained from 92 women referred for abnormal Pap smears. The lavages and one of the biopsies were snap-frozen and subsequently tested by SBH for HPV DNA. The other biopsy was formalin-fixed and paraffin-embedded for conventional light microscopy and for ISH using two commercially available kits from Digene Diagnostics Inc.: ViraType In Situ containing probe mixtures to HPVs 6 and 11, 16 and 18, and 31, 33, and 35; and Omniprobe containing a single probe mixture to 14 different anogenital HPVs. In addition to the aforementioned types Omniprobe contains probes for types 42, 43, 44, 45, 51, 52, and 56. HPV DNA was detected by SBH in 27% of specimens showing no specific pathological change and in 92% of cases of cervical intraepithelial neoplasia (CIN). The HPVs by SBH in CIN were type 16 in 25%, types 31 or 35 in 23%, types 52 or 56 in 17%, uncharacterized types in 13%, mixed types in 8%, and type 18 in 6%. None of the specimens was positive for the low-risk HPV types 6, 11, 42, 43, or 44. The proportion of cases of CIN containing HPV 16 increased as the grade of CIN increased. Otherwise there were no striking differences detected in the HPV types in the different grades of CIN. ISH using ViraType In Situ was positive in only 50% of cases of CIN that were HPV positive by SBH.(ABSTRACT TRUNCATED AT 250 WORDS)