The role of single nucleotide polymorphisms of the ERCC1 and MMS19 genes in predicting platinum-sensitivity, progression-free and overall survival in advanced epithelial ovarian cancer.

OBJECTIVE: This study aims to assess the role of polymorphisms in DNA repair genes, excision repair cross-complementation group 1 (ERCC1) and methyl-methanesulfonate sensitivity 19 (MMS19), in tumor response to platinum-based chemotherapy and survival in advanced epithelial ovarian cancer (EOC). METHODS: Single nucleotide polymorphism (SNP) analysis was performed on the paraffin-embedded tumor tissue of women with advanced EOC, treated with platinum-based chemotherapy at the University of Oklahoma Health Sciences Center. Polymorphisms from two ERCC1 (codon-118 and C8092A) and three MMS19 (rs2211243, rs2236575 and rs872106) gene loci were evaluated by real time PCR Allelic Discrimination Assay. RESULTS: Genotyping was performed in 107 patients, 45 platinum-sensitive and 62 platinum-resistant. ERCC1, codon-118 and C8092A genotyping was evaluable in 98 and 106 patients respectively and in all 107 patients for MMS19 polymorphisms. No differences were observed in genotype between platinum-sensitive and platinum-resistant patients. Polymorphisms in the ERCC1, codon-118 and MMS19 genes did not correlate with overall survival (OS), although a trend toward improved progression free survival (PFS) was observed in patients expressing the minor (GG) alleles of the rs872106 MMS19 gene. Women homozygous for the ERCC1-C8092A minor (AA) alleles had a significant increase in PFS compared to AC and CC patients and both AA and AC genotypes conferred improved survival over the major (CC) genotype. CONCLUSIONS: Polymorphisms in ERCC1, codon-118 and MMS19 genes are not associated with clinical response to platinum or survival. The ERCC1-C8092A genotypes containing an "A" allele were associated with significant improvement in PFS and OS strengthening the value of this specific genotype in survival.

Wnt inhibitory factor 1 induces apoptosis and inhibits cervical cancer growth, invasion and angiogenesis in vivo.

Aberrant activation of Wingless-type (Wnt)/ß-catenin signaling is widespread in human cervical cancer. However, the underlying mechanisms of Wnt activation and the therapeutic potential of Wnt inhibition remain largely unknown. Here, we demonstrate that the Wnt inhibitory factor 1 (WIF1), a secreted Wnt antagonist, is downregulated in all human primary cervical tumors and cell lines analyzed. Our data reveal that WIF1 downregulation occurs due to promoter hypermethylation and is an early event in cervical oncogenesis. WIF1 re-expression upon 5-aza-2'-deoxycytidine treatment or WIF1 gene transfer induces significant apoptosis and G(2)/M arrest, and inhibits cervical cancer cell proliferation in vitro. Consistent with this, treatment of established mice tumor xenografts with peritumoral WIF1 gene transfer results in a significant inhibition of cancer growth and invasion. WIF1 treatment causes a significant decrease in intracellular WNT1 and TCF-4 proteins revealing novel Wnt-regulatory mechanisms. Thus, WIF1 causes a major cellular re-distribution of ß-catenin and a significant inhibition of the Wnt/ß-catenin pathway in tumor cells, as documented by a remarkable reversion in the expression of Wnt/ß-catenin transcriptional target genes (E-cadherin, c-Myc, cyclin D1, CD44 and VEGF). Consequently, multiple critical events in tumor progression and metastasis such as cell proliferation, angiogenesis and invasion were inhibited by WIF1. In addition, WIF1 modulated the expression of specific anti-apoptotic and apoptotic proteins, thereby inducing significant apoptosis in vivo. Our findings demonstrate for the first time that WIF1 downregulation by epigenetic gene silencing is an important mechanism of Wnt activation in cervical oncogenesis. Of major clinical relevance, we show that peritumoral WIF1 gene transfer reduces not only cancer growth but also invasion in well-established tumors. Therefore, our data provide novel mechanistic insights into the role of WIF1 in cervical cancer progression, and the important preclinical validation of WIF1 as a potent drug target in cervical cancer treatment.

Evaluation of linear array human papillomavirus genotyping using automatic optical imaging software.

Variations in biological behavior suggest that each carcinogenic human papillomavirus (HPV) type should be considered individually in etiologic studies. HPV genotyping assays might have clinical applications if they are approved for use by the FDA. A widely used genotyping assay is the Roche Linear Array HPV genotyping test (LA). We used LA to genotype the HPV isolates from cervical specimens from women with the full spectrum of cervical disease: cervical cancer, cervical intraepithelial neoplasia (CIN), and HPV infections. To explore the feasibility and value of the automated reading of the LA results, we custom-designed novel, optical imaging software that provides optical density measurements of LA bands. We compared unmagnified visual examination with the automated measurements. The two measurements were highly associated. By either method, the threshold between a negative and a positive result was fairly sharp, with a clear bimodal distribution. Visually, most positive results were judged to be strong or medium, with fewer equivocal results categorized as weak (9.5% of positive samples), very weak (6.5% of positive samples), or extremely weak (7.7% of positive samples). The automated measurements of the intensities were significantly associated with the strength of the visual categories (P < 0.001). At the extremes of the automated signal intensities (< or = 20 units or > or = 120 units), the bands were almost always categorized visually as negative and positive, respectively. In the equivocal zone (20 to 119 units), specimens were more increasingly likely to be judged to be visually positive as the number of other, definite infections on the same strip increased (P for trend < 0.001). Multiple, concurrent infections comprise > or = 25% of HPV infections; thus, any systematic visual tendency that influences their evaluation when the result is equivocal should be minimized. Therefore, automated reading is probably worth development if easy-to-calibrate hardware and software can be optimized.

HPV DNA testing of the residual sample of liquid-based Pap test: utility as a quality assurance monitor.

HPV DNA testing of the residual sample volume of liquid-based Pap tests has been recommended as a way to determine the appropriate follow-up for women who have equivocal results in routine clinical screening. A major aspect of quality assurance in the cytopathology laboratory consists of correlation of smear interpretation with biopsy or conization results as mandated by CLIA '88. However, the use of histology as the gold standard suffers from similar problems of subjectivity and sampling as the Pap smear. In this study we explore the potential use of HPV DNA testing of the residual volume from the ThinPrep Pap Test (Cytyc Corporation, Boxborough, Massachusetts) as a substitute gold standard in quality assurance monitoring of a cervical cytology screening program. The residual samples from 397 ThinPrep Pap cases were retrospectively analyzed for high-risk HPV DNA using the Hybrid Capture II technique. Sensitivity (71.8%), specificity (86.5%), predictive value of positive (77.1%) and negative (82.9%) ThinPrep Pap interpretations were calculated on the basis of HPV DNA results for 266 cases classed as either squamous intraepithelial lesion (SIL) or negative. Overall, there was agreement between the two tests in 80.8% of cases (Cohen's kappa =.59). The percentage of HPV DNA-positive cases interpreted as atypical squamous cells of uncertain significance (ASCUS) was 43.7%, and the percentage of negative cases was 17.1%. We believe that this approach is an objective adjunct to the traditional quality assurance protocol, with the added benefit that it includes cases interpreted as negative, as well as abnormal cases that do not come to biopsy.

The significance of allelic deletions and aneuploidy in colorectal carcinoma. Results of a 5-year follow-up study.

BACKGROUND: A prospective study was initiated to analyze the prognostic value of both the deletion of candidate tumor suppressor genes and the DNA content in colorectal carcinoma specimens. METHODS: A prospective study was initiated in 1988, into which 104 patients from the Brooklyn VA Medical Center were accrued through March 1992. DNA restriction endonuclease digests, obtained by the Southern blot technique, were examined for allelic deletions by cDNA probes pnm23-H1 (17q21) YNZ 22.1 (17p13), and p15-65 (18q21). DNA content was measured by image analysis cytometry of Feulgen-stained tumor imprints. Median follow-up was 5.5 years (range, 4-8.5 years). RESULTS: Patients with nm23-H1 allelic deletions were 3 times as likely to develop distant metastases as patients without nm23-H1 deletions (relative risk [RR], 3.89; 95% confidence interval [CI], 1.39, 10.89). This connection was even stronger after adjustment for TNM stage and site of primary tumor (RR, 5.27; 95% CI, 1.67, 16.68). No significant association of 17p or 18q deletions or ploidy with either distant metastases or overall survival was noted. In multivariate analysis, clinicopathologic variables associated with decreased survival included intracellular mucin production, nuclear grade, TNM stage, and nerve invasion. CONCLUSIONS: A combination of clinicopathologic and molecular biologic variables may identify patients at high risk for death from disseminated colorectal carcinoma.

Malignant struma ovarii. A case report and review of the literature.

Malignant struma ovarii is a rare form of ovarian carcinoma; only 3 cases of its pure papillary type have been reported in the literature. A new case of malignant papillary struma ovarii arising in an asymptomatic 32-year-old woman is presented. Due to its rarity, there has been confusion in the diagnosis and management of malignant struma ovarii. Criteria for the diagnosis of malignant papillary struma ovarii are proposed. Conservative treatment after a complete staging procedure is possible due to the usually benign course and low incidence of metastases of this tumor.

Utility of the cervical cytologic smear in assessing endocervical involvement by endometrial carcinoma.

OBJECTIVE: To evaluate the possible contribution of cervical cytology in the identification of high-risk endometrial cancer patients. STUDY DESIGN: A retrospective study of 61 patients who had a preoperative cervical cytologic smear and hysterectomy at our institution for endometrial carcinoma. The smear and endocervical curetting (ECC) results were compared with the status of the endocervix in the hysterectomy specimens. RESULTS: Two patterns of malignant endometrial cells were identified in the 25 positive smears: (1) a sloughing pattern, which was the classic rounded cell pattern associated with the exfoliation of endometrial cancer cells, and (2) an abraded pattern in which the cancer cells were present as loosely cohesive, sheetlike groups that retained the original cell shape. This pattern was associated with endocervical involvement by endometrial cancer and overlapped with the criteria for primary cervical adenocarcinoma. Using the histologic status of the endocervix in the hysterectomy specimen as the standard, cervical cytology compared favorably with ECC in predicting the status of the endocervix. Pitfalls included bulky or polypoid lesions that abutted the endocervical canal and smears taken when the endometrium was sloughing. Reactive endocervical cells presented diagnostic dilemmas in patients who had had endometrial sampling prior to the smear. When restricted to cases in which the smear preceded endometrial sampling, the smear was superior to ECC in predicting endocervical involvement. CONCLUSION: These results suggest that preoperative smears may be valuable in assessing cervical involvement by endometrial carcinoma. It is recommended that a smear be performed as an initial procedure in any woman with complaints of abnormal uterine bleeding.

Peritoneal washing cytology in gynecologic cancers: long-term follow-up of 355 patients.

BACKGROUND: Microscopic evaluation of cells washed from the peritoneal cavity during surgery for gynecologic tumors is used to detect subclinical intraperitoneal metastases from these tumors. The prognostic significance of this test, however, has been questioned. PURPOSE: Stressing histologic correlation and pitfalls in interpretation, we previously reported that the sensitivity of intraoperative peritoneal washing cytology was lower than was suggested earlier. This study evaluates the clinical utility of this test in the long-term follow-up of our patients. METHODS: Staging (International Federation of Gynecology and Obstetrics [FIGO], 1971) and follow-up information was available for 355 unselected patients with primary tumors who had peritoneal washings performed during initial surgery at University Hospital-Stony Brook, NY, during the period from 1980 through 1989. There were 135 patients with endometrial carcinomas, 112 with ovarian carcinomas, 92 with cervical carcinomas, and 16 with borderline (i.e., of low malignant potential) ovarian tumors. The median follow-up of the patients was 57 months (range, 0-154 months). Follow-up data were obtained from the Tumor Registry at University Hospital-Stony Brook. Survival differences were determined by Kaplan-Meier analysis and were evaluated by two-tailed logrank test. RESULTS. Peritoneal washing cytology was positive at initial surgery for 120 (33.8%) of 355 patients, including 90 (80.4%) of 112 patients with ovarian carcinomas, five (31.2%) of 16 patients with borderline ovarian tumors, 17 (12.6%) of 135 patients with endometrial carcinomas, and eight (8.7%) of 92 patients with cervical cancers. For 203 patients with stage I tumors, the peritoneal cytology was positive in 29.4% of the patients with ovarian carcinomas, 18.2% with borderline ovarian tumors, 6.1% with endometrial carcinomas, and 5.2% with cervical carcinomas. By use of peritoneal histology as the standard, peritoneal cytology was highly specific (98.1%) but less sensitive (82.9%) in detecting intraperitoneal involvement. For patients with stage I tumors, 80.0% with ovarian carcinomas, 83.3% with endometrial carcinomas, and 100% with cervical carcinomas who showed positive cytology died of their cancer, compared with 25.0% with ovarian carcinomas, 13.0% with endometrial carcinomas, and 21.9% with cervical carcinomas who showed negative peritoneal cytology. Four (2.0%) patients with stage I tumors had positive peritoneal cytology but negative peritoneal histology. Of these patients, three (two with ovarian carcinoma and one with cervical carcinoma) died of their cancer, whereas one patient with a borderline ovarian tumor was free of disease at the last follow-up. Survival analysis indicated that peritoneal washing cytology stratified for stage provides better prognostic information for each primary cancer site studied than does stage alone. All patients with borderline ovarian tumors were alive at last follow-up, regardless of disease stage or peritoneal status. CONCLUSIONS: Regardless of FIGO stage, positive peritoneal washing cytology predicted poor prognosis for women with epithelial tumors of the genital tract, except for patients with borderline ovarian tumors. Patients in whom peritoneal cytology was the only evidence of intraperitoneal spread were few, but the disease in such patients was associated with poor outcome. IMPLICATIONS: Strict adherence to specialized cytologic criteria in peritoneal washing cytology allows for results that are highly predictive of survival. This information may be useful in stratifying women in therapeutic trials for treatment of genital tract carcinomas.

Primary intratesticular sarcoma. Immunohistochemical ultrastructural and DNA flow cytometric study of three cases with a review of the literature.

BACKGROUND: Fourteen cases of testicular sarcoma have been reported in the contemporary era. These included 7 cases of rhabdomyosarcoma, 2 spindle cell sarcoma, 2 osteosarcoma, 1 leiomyosarcoma, 1 fibrosarcoma, and 1 chondrosarcoma coma. METHODS: In this report, immunohistochemical stains, electron microscopy, and DNA flow cytometric analysis were performed on primary testicular sarcomas from three patients. RESULTS: The patients were age 47, 40, and 33 years. Each presented initially with a testicular mass. The tumors measured 4.8, 4.0, and 4.0 cm in greatest dimension. There was no associated germ cell elements nor elevated alpha-fetoprotein or beta-human chorionic gonadotropin. Case 1 was positive for actin, vimentin, and alpha-1-chymotrypsin. Case 2 was positive for vimentin but not desmin. Case 3 was positive for desmin and S-100. Smooth muscle differentiation was identified by electron microscopy. Flow cytometric analysis revealed DNA aneuploidy in all cases: 1.27, 1.29, and 1.71. The 3 patients were alive and well without recurrent disease at 7, 6, and 4 years after diagnosis. Inguinal orchiectomy was the initial treatment in all 17 patients, there was 1 death from metastatic disease and 2 patients with distant metastases. CONCLUSION: Primary testicular sarcoma is a rare indolent tumor with potential for distant metastases. Two cases of primary testicular leiomyosarcoma and one of unclassified sarcoma of the testis are reported.

Mucinous adenocarcinoma of the endometrium with intestinal differentiation: a case report.

Mucinous differentiation of endocervical type has been well documented in endometrial carcinoma. However, we describe an unusual case of adenocarcinoma of the endometrium showing diffuse histological, immunohistochemical, and ultrastructural evidence of intestinal differentiation. Although intestinal differentiation has been described in mesodermally derived tissues including endocervix, ovary, and urinary tract, it has not been reported in normal endometrium. One previous case has been reported showing this pattern in endometrial carcinoma. Possible histogenetic mechanisms of this pattern are discussed.

Peritoneal washing cytology in cervical carcinoma. Analysis of 109 patients.

The peritoneal washing cytologies of 109 patients (112 procedures) undergoing laparotomy for cervical carcinoma were evaluated retrospectively and compared with the clinical and pathologic findings. Nine patients (8.3%) had malignant peritoneal washings (including three of four with washings initially termed "inconclusive"). Four (4.9%) of the 82 patients with squamous carcinoma and 3 (16.7%) of 18 with adenocarcinoma had positive washings. Five (5.6%) of 90 washings obtained at initial explorations were positive, as compared with 4 (18.2%) of 22 washings obtained as follow-up operations in recurrent cases. The 111 peritoneal washing cytologies with a corresponding histologic evaluation of the peritoneal cavity showed a good correlation; peritoneal washing cytology had an efficiency of 91.0%, a sensitivity of 52.9% and a specificity of 100%. Two cases in which the cytologies were considered positive only after review had negative peritoneal histologies; both patients died of progressive disease within 11 months. Peritoneal washing cytology was positive in 5 (5.9%) of 84 cases with FIGO stage 1 cancers, 2 (18.2%) of 11 cases with stage 2 cancers, 1 (33.3%) of 3 cases with stage 3 cancers, and 1 (10%) of 10 cases with recurrent tumors. Eight (88.9%) of nine patients with malignant peritoneal washings died of disease from 3 to 15 months following surgery; one showed no evidence of disease at 9 months. These results suggest that: (1) cervical carcinomas are infrequently associated with a positive peritoneal washing; (2) peritoneal washing cytology is more likely to be positive in cases of adenocarcinoma than in cases of squamous carcinoma; (3) peritoneal washings obtained at the time of surgery for recurrence are more likely to contain malignant cells than are washings obtained during initial exploration; (4) nonkeratinizing malignant squamous cells may be confused with reactive mesothelial cells; and (5) peritoneal washing cytology is a relatively insensitive technique for detecting advanced cervical disease, but correlates with a poor prognosis when positive.

Assessment of criteria used in the histologic diagnosis of human papillomavirus-related disease of the female lower genital tract.

The goal of this study was to evaluate the histologic criteria used to establish the diagnosis of human papillomavirus (HPV)-associated disease, especially in borderline lesions. In a completely blinded study, 21 patients had one biopsy each of the cervix and vulva. Each specimen was evaluated by RNA and DNA in situ hybridization, a histologic diagnosis was rendered, and then each was evaluated for 12 histologic criteria commonly associated with HPV. On the cervix only binucleation and dysplasia correlated well with in situ hybridization. Koilocytosis correlated very strongly with the histologic diagnosis. On the vulva, koilocytosis, papillomatosis, elongated rete pegs, binucleation, and hypergranulosis correlated well with in situ hybridization. When four other pathologists reviewed the slides, they agreed on the histologic diagnosis and the presence of koilocytosis, binucleation, and dysplasia on the cervix but on none of the other criteria. On the vulva the pathologists disagreed on the overall diagnosis and the presence of any of the criteria with the exception of papillomatosis. Nonclassic histologic criteria should not, by themselves, be used to make the diagnosis of condyloma. The use of such terminology as "suggestive of condyloma" in histologic diagnoses should be avoided in favor of more descriptive terminology to avoid possibly unnecessary treatment for lesions of questionable significance.

Cytohistologic correlation of peritoneal washing cytology in gynecologic disease.

The peritoneal washings and cul de sac aspirates from 204 patients undergoing 217 procedures for the evaluation of gynecologic disease were examined retrospectively and correlated with the histologic diagnoses. Of the 73 washings from patients with histologically benign genital disease, cytology diagnosed 64 (87.7%) as negative, 6 (8.2%) as inconclusive and 3 as malignant. One malignant washing was a true positive from a nongenital primary. False positives thus occurred in 2.7% of the benign cases on blind review. Of 144 cytologic examinations of washings from patients with histologically confirmed malignant disease of the female genital tract, 38 (26.4%) were considered positive after cytohistologic correlation. Four malignant cases (2.1%) were undercalled on blind review while 3 (2.0%) were considered overcalls. Eleven of 47 cases (23.4%) with biopsy-proven peritoneal disease had negative cytology after histologic correlation. Recurring problems in interpreting peritoneal washings included: (1) the differential diagnosis of the spectrum encompassed by reactive mesothelium, endosalpingiosis, borderline serous tumors and well-differentiated serous cystadenocarcinoma; (2) morphologic similarities between some tumor cells and mesothelial cells associated with treatment effects; and (3) a paucity of malignant cells in some washings, resulting in false-negative interpretations. Ineffective cytopreparation, particularly of bloody specimens, hampered interpretation of some specimens. Correlation with previous histology and cytology enhanced the accuracy of peritoneal washing cytology in this study.

Hepatocellular carcinoma presenting as an oral cavity lesion.

A primary hepatocellular carcinoma associated with underlying hemochromatosis appeared as a metastatic mandibular lesion. A swelling of the retromolar area and mandibular paresthesia were the presenting complaints. Hepatocellular carcinoma rarely metastasizes to the orofacial area. To our knowledge, only 18 cases have been reported. The clinical, radiographic, and histologic features of an additional case of hepatocellular carcinoma metastatic to the left posterior alveolar ridge is reported, and the literature is reviewed.

Candida pericarditis and tamponade in a patient with systemic lupus erythematosus.

Candida pericarditis and tamponade developed in a patient with sterile purulent pericarditis secondary to systemic lupus erythematosus. Therapy with amphotericin B and properly timed surgical intervention led to a clinical and microbiological cure. This article emphasizes the importance of differentiating an infected pericardial effusion from the sterile pericarditis of systemic lupus erythematosus and provides suggested guidelines for the management of that complication.

Cytologic findings in peritoneal washings associated with benign gynecologic disease.

In order to define the problems in the interpretation of peritoneal washing cytology specimens, 149 benign cases were examined retrospectively and the cytologies compared with the correlating histologies. There were problems in the cytologic interpretation on blind review in 18 cases (12.1%). Difficulties in interpretation of peritoneal washing cytology included conditions associated with (1) mesothelial reactions and adhesion formation, (2) rupture of cystic structures, (3) fallopian tube epithelium and (4) combinations of these factors. It is anticipated that familiarity with these problems will enhance accuracy in peritoneal washing cytology, particularly in those cases in which a malignancy is present but has not penetrated into the peritoneal cavity.