Cases Of Invasive Adenocarcinoma Research Articles

Prevalence of human papillomavirus types 16 and 18 in cervical adenocarcinoma and its precursors in Scottish patients

Our aim was to determine the prevalence of human papillomavirus (HPV) types 16 and 18 in cervical adenocarcinoma (and its precursors) in Scottish patients. Nucleic acid was extracted from paraffin-embedded, formalin-fixed tissues. We examined 119 cases of invasive adenocarcinoma, 20 cases of adenocarcinoma in situ, and 16 cases of normal glandular epithelium. HPV DNA was detected by polymerase chain reaction using type-specific primers for the E6 and E7 genes of HPV-16 and HPV-18 with conformation of HPV genotype by subsequent restriction fragment length polymorphism. HPV DNA was identified in 87 (62.6%) cases, with HPV-16 being detectable in 65 (47%) cases and HPV-18 in 41 (29%) cases. All the cases of normal tissue tested negative for HPV-16 and/or HPV-18. No significant relation between infecting HPV type (16 or 18) and subtypes of disease (within the invasive category and between the preinvasive and the invasive categories) was noted. Our findings support that HPV-16, along with HPV-18, are likely to play a significant role in the pathogenesis of cervical adenocarcinomas and that cervical cancer screening strategies that incorporate oncogenic HPV testing, and prophylactic vaccines that target these types, will be beneficial for the reduction of adenocarcinoma and associated glandular precursors.

Evaluation ofPTENexpression 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. SomaticPTENmutations are involved in a variety of tumors, including endometrial carcinomas, wherePTENexpression is diminished. We examined expression ofPTENin 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-μm sections were cut and immunostained withPTEN, and expression was correlated with clinicopathologic variables, including histologic subtypes of adenocarcinoma. In 137 patients,PTENexpression was positive in 121 (88%). The intensity and distribution ofPTENstaining in the tumor tissue were more heterogeneous than those observed in the normal tissues. There were no significant differences in distribution or intensity ofPTENexpression between adenocarcinomain situand subtypes of invasive adenocarcinoma. Our findings show that unlike the case in most endometrial carcinomas,PTENexpression is retained during the process of carcinogenesis in the glandular cervix. There is, however, evidence of altered distribution and intensity ofPTENexpression in cervical adenocarcinoma cells.

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.

Prevalence of human papillomavirus types 16 and 18 in cervical adenocarcinoma and its precursors in Scottish patients

Our aim was to determine the prevalence of human papillomavirus (HPV) types 16 and 18 in cervical adenocarcinoma (and its precursors) in Scottish patients. Nucleic acid was extracted from paraffin-embedded, formalin-fixed tissues. We examined 119 cases of invasive adenocarcinoma, 20 cases of adenocarcinoma in situ, and 16 cases of normal glandular epithelium. HPV DNA was detected by polymerase chain reaction using type-specific primers for the E6 and E7 genes of HPV-16 and HPV-18 with conformation of HPV genotype by subsequent restriction fragment length polymorphism. HPV DNA was identified in 87 (62.6%) cases, with HPV-16 being detectable in 65 (47%) cases and HPV-18 in 41 (29%) cases. All the cases of normal tissue tested negative for HPV-16 and/or HPV-18. No significant relation between infecting HPV type (16 or 18) and subtypes of disease (within the invasive category and between the preinvasive and the invasive categories) was noted. Our findings support that HPV-16, along with HPV-18, are likely to play a significant role in the pathogenesis of cervical adenocarcinomas and that cervical cancer screening strategies that incorporate oncogenic HPV testing, and prophylactic vaccines that target these types, will be beneficial for the reduction of adenocarcinoma and associated glandular precursors.

Prognostic factors and clinicopathologic characteristics of invasive adenocarcinoma of the uterine cervix

The purpose of this study was to evaluate the outcome of patients with cervical adenocarcinoma and to determine the characteristics and the prognostic factors of this entity. This retrospective study was done in the Department of Surgical Oncology of the Salah Azaiz Institute of Tunis with 79 cases of invasive adenocarcinoma of the uterine cervix that were collected from 1990 to 1999. Survival was analyzed according to the Kaplan-Meier method. Univariate analysis of prognostic factors was performed with the test of log rank. Cox regression model was used in multivariate analysis of prognostic factors. Mean age was 50 years, and metrorrhagia was mostly revealing in 73% of the cases. Early stages (I, IIa, IIb with 1/3 proximal parametrial invasion) and "pure" type adenocarcinoma were found in 78% and 87% of the cases, respectively. Treatment consisted of a radiosurgical combination in 52 cases; exclusive radiotherapy was practiced with 17 patients. The 5 year-overall and disease-free survival percentages were, respectively, 68% and 72.4%. Poor prognostic factors were age >50 years, tumor size >4 cm, advanced stage, tumor grade, and lymph nodes and lymph-vascular space involvement. With the use of multivariate analysis, only stage and lymph node metastases remained significant prognostic factors. This report shows survival and prognostic factors that are similar to those found in previous studies, but unlike the Western countries, our results demonstrate a high rate of early stages and no increase in frequency of cervical adenocarcinoma.

A case of invasive adenocarcinoma derived from intraductal papillary-mucinous carcinoma showing high MUC 1 expression in non-invasive component: Possible relationship between MUC 1 expression and tumor invasion

A case of invasive adenocarcinoma derived from intraductal papillary-mucinous carcinoma showing high MUC 1 expression in non-invasive component: Possible relationship between MUC 1 expression and tumor invasion

Glandular Lesions of the Cervix on Thin-Layer Pap Tests

To determine the cytologic features that are most helpful in characterizing significant glandular lesions of the cervix observed on the ThinPrep (TP) Pap test (Cytyc Corp., Boxborough, Massachusetts, U.S.A.) and to compare these features with those published for conventional smears. Thirty-nine TP preparations with cytologic evidence of glandular lesions of the cervix and histologic and/or clinical correlation were studied. These lesions included (1) 11 cases of benign/reactive conditions; (2) 10 cases of adenocarcinoma in situ (AIS), of which 1 had both AIS and carcinoma in situ; (3) 1 case of invasive adenocarcinoma; (4) 15 cases of squamous intraepithelial lesions and squamous cell carcinoma, including 4 with glandular involvement, and (5) 2 cases of adenosquamous cell carcinoma. These cases were reviewed by the first author without knowledge of the histologic diagnosis. Twenty-five previously published cytologic criteria were used to evaluate glandular cells on TP slides. Statistical analysis was performed using Fisher's exact test to determine the significance of the features studied. All glandular lesions had cytologic features on TP similar to those previously described on conventional smears. However, TP slides demonstrated enhanced nuclear features but less-preserved architectural patterns. Reactive lesions showed minimal overlapping without hyperchromasia or mitotic figures and with normal nuclear/cytoplasmic ratios. AIS and invasive adenocarcinoma cases had similar features. Increased cellularity and overcrowding were prominent, whereas feathering, rosettes and cell strips were present but subtle. Glandular lesions of the cervix on TP slides shared many of the characteristic features reported for conventional smears. However, nuclear details were more pronounced in TP slides, while architectural patterns, although present, were relatively subtle.

Proliferation to apoptosis ratio as a prognostic marker in adenocarcinoma of uterine cervix

Objectives. To determine the Mitotic Index (MI), Apoptotic Index (AI), the ratio of the two indices (MI/AI) in normal endocervical glands, adenocarcinoma in situ (AIS) and invasive adenocarcinoma of cervix, and to evaluate the relationship among these indices with various clinicopathological features. Methods. The MI, AI and MI/AI ratio in cervical adenocarcinoma were evaluated based on: (1) cell morphology in hematoxylin and eosin-stained sections; (2) immunohistochemical study for Ki67 antigen and terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL). Eighty cases of invasive adenocarcinoma and eighteen cases of adenocarcinoma in situ (AIS) adjacent to invasive adenocarcinoma were included. Adjacent normal endocervical epithelium in 26 cases of adenocarcinoma was included as control. Results. The MI, AI and MI/AI in normal endocervical glands, AIS and invasive carcinomas showed statistically significant differences ( P < 0.001). A significant positive correlation was found between AI and MI, as assessed by morphological features ( P < 0.001) and immunohistochemistry ( P = 0.006). The MI/AI ratio, determined by morphology, significantly correlated with staging ( P = 0.023) and survival ( P = 0.0045). Multivariate survival analysis showed that both MI/AI ratio determined by morphology ( P < 0.001) and stage of tumor ( P = 0.03) had independent prognostic value in invasive adenocarcinoma. Conclusions. Tumor proliferation significantly correlated with apoptotic activity in cervical adenocarcinoma. The MI/AI ratio was an independent prognostic factor associated with patient survival. Histological determination of MI/AI ratio proved to be an economical and potentially useful adjunct in predicting clinical outcome of patients with cervical adenocarcinoma.

Altered Expression of β-Catenin during Radiation-induced Colonic Carcinogenesis

Radiotherapy for malignant pelvic disease is commonly accompanied by treatment-induced proctitis, and rarely by colorectal cancer. Translocation of the beta-catenin protein, which is a key downstream effector of the Wnt signal transduction pathway, is frequently found in colorectal cancer. Nuclear beta-catenin enhances transcriptional activity of the cyclin D1 gene in cancer cells. Here, we evaluate the involvement of the Wnt pathway in radiation-induced colon carcinogenesis with rats (n = 36). Beta-catenin, APC, and cyclin D1 expression profiles were analyzed by immunohistochemistry in radiation-induced chronic colon injury including cancers and ulcerative lesions in rats (n = 12 in treated group, n = 12 in control group). In total, 3 cases of invasive adenocarcinomas were developed in the irradiated portion 50 weeks after a single dose of 36 Gy irradiation. Nuclear translocation of beta-catenin was observed in all radiation-induced colon cancers, whereas this protein was also found in the cytoplasm and/or nucleus of 9 cases of non-neoplastic irradiated colonocytes. Nuclear translocation of beta-catenin correlated with loss of APC and gain of cyclin D1 expression, suggesting activation of the Wnt pathway during radiation-induced colorectal carcinogenesis. A single dose of 10 Gy was also given for acute injury (n = 12: 3 each in days 0, 3, 5, and 7, respectively). Beta-catenin expression was distributed in the cytoplasm of degenerating glands at day 3 and 5, and was observed in the cell membrane of those glands with histological normalization at day 7 after irradiation. Because translocation of beta-catenin was found in irradiated-colonic mucosa as well as colon cancer, disruption of beta-catenin expression might be one of the early events in radiation-induced colonic carcinogenesis.

Expression of Matrix Metalloproteinases in Invasive Pulmonary Adenocarcinoma with Bronchioloalveolar Component and Atypical Adenomatous Hyperplasia

To investigate the association between the expression of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) and the clinicopathological features in lepidic and invasive components of adenocarcinoma of the lung, we performed immunostaining for type IV collagen, various MMPs, and TIMPs in 27 cases of invasive adenocarcinomas and 5 cases of atypical adenomatous hyperplasia of alveolar epithelial cells (AAH). Mean extent of lepidic growth was 61% and the survival was significantly better in cases with 50% or more lepidic component. The preservation of type IV collagen in lepidic areas correlated inversely with lymphatic or vascular invasion (P = 0.02 and 0.002, respectively). Five-year survival was reduced in cases showing destruction of type IV collagen (P = 0.004) or expression of MMP-2 (P = 0.008) in lepidic areas. MMP-2 co-localized with MT-1-MMP (its activating enzyme) and TIMP-2 in neoplastic cells. Reactivity for other MMPs and TIMPs did not correlate with destruction of type IV collagen or prognosis. Type IV collagen was preserved in all cases of AAH. MMP-2, but not MT-1-MMP, was expressed in two of the five cases of AAH. Immunostaining for type IV collagen MMP-2 is useful in evaluating the prognosis of the lung.

Altered Expression of CD44 and Variant Isoforms in Human Adenocarcinoma of the Endocervix during Progression

Objective. Altered expression of the CD44 family of cell adhesion molecules has been associated with tumor progression and metastasis. The aim of this study was to investigate the expression of the gene products of CD44 standard (CD44s) and several alternatively spliced variants (CD44v4, v6, v7, and v9) in adenocarcinoma of the endocervix and to correlate the degree of their expression with disease progression.Methods. Immunohistochemical staining for CD44s and CD44v4, v6, v7, and v9 was performed on formalin-fixed, paraffin-embedded endocervical specimens. Seventeen cases of adenocarcinoma in situ (AIS) and 22 cases of invasive adenocarcinoma of the endocervix were included in this study, and the immunoreactivity was compared with that of normal endocervical epithelium.Results. (1) In the normal endocervical mucosa, immunoreactivity for CD44s and the splice variants was lacking or was confined to only the basal portion of the glandular epithelium along the basement membrane; (2) CD44s was diffusely expressed along the entire cytoplasmic membrane, including the luminal surface of the tumorous glands in 94% of AIS and 95% of invasive adenocarcinomas; (3) a significantly stronger expression of CD44s was observed in invasive adenocarcinomas than in AIS; (4) in contrast to all other splice variants, CD44v9 demonstrated an increased expression in nearly all in situ and invasive lesions compared to the normal tissue; (5) CD44v4 and v6 were expressed only in a small proportion of invasive adenocarcinomas and were near totally absent in the in situ lesions; and (6) CD44 v7 was totally absent in all normal, in situ, and invasive lesions studied.Conclusions. It appears that neoplastic transformation of endocervical epithelium is associated with qualitative and quantitative changes in the expression of CD44 standard molecule and some CD44 splice variants.

Endocervical Glandular Neoplasia and Its Mimics in ThinPrep® Pap Tests

To document the cytologic features of endocervical adenocarcinoma in situ (AIS) and invasive endocervical adenocarcinoma as observed in ThinPrep slides and to compare these features with those that have been described for conventional smears. Six cases of endocervical AIS and three cases of invasive adenocarcinoma were evaluated with respect to 3 low-power and 14 high-power features. All cases were biopsy proven. Glandular "look-alikes" (tubal metaplasia, n = 3; florid repair, n = 3; sampling of lower uterine segment, n = 1) were also examined. All cases of AIS contained dark groups and sheets at screening power. At higher power, nuclear detail was extremely well visualized. All cases had crowding, continuous depth of focus, variability of nuclear size and shape within groups, irregular nuclear membranes, uniformly stippled chromatin and at least occasional single atypical cells. Only one case lacked nucleoli. Traditional features (strips, feathering, rosettes and mitoses) were observed about as frequently as in conventional smears. Invasive lesions had many of the same features, with relatively more inflammation and lysed blood. Nonneoplastic look-alikes could be distinguished from neoplasms using traditional criteria. In this small study, AIS and invasive endocervical adenocarcinoma maintained the features previously described for conventional smears. Improved visualization of nuclear detail may allow the application of additional criteria, such as irregular nuclear membranes and the more consistent presence of nucleoli, for distinguishing glandular neoplasms from their look-alikes.

The management and follow-up of patients with high-grade cervical glandular intraepithelial neoplasia

Maini M, Lavie O, Comerci G, Cross PA, Bolger B, Lopes A, Monaghan JM. The management and follow-up of patients with high-grade cervical glandular intraepithelial neoplasia. Int J Gynecol Cancer 1998; 8: 287–291. The purpose of this study was to evaluate if the status of the margins on large loop excision of the transformation zone (LLETZ) can predict the presence of residual cervical glandular intraepithelial neoplasia (CGIN) and to assess the efficacy of cervical conization as primary management of CGIN. Between January 1989 and March 1997 fifty patients with a diagnosis of CGIN made by LLETZ were treated in the Department of Gynaecological Oncology at the Queen Elizabeth Hospital, Gateshead. Presence or absence of CGIN at cone margins, results of cervical cytological examinations before and following conization, colposcopic examination, and results of histopathologic assessment of any surgical specimens taken after initial cone biopsy were analyzed in this study. Of the 50 patients with CGIN, managed by conization, in 34 (68%) CGIN was present at the surgical margins and in 16 (32%) the margins were clear. Eleven (32.3%) patients with CGIN at the margins of excision had evidence of residual disease and there was only one of the 16 (6.25%) patients with clear margins (P= 0.03). Two cases of invasive adenocarcinoma were identified in the group of patients with positive margins. In a mean follow-up time of 32.35 months, no case of invasive carcinoma was identified in the completely excised group. In our retrospective study LLETZ was found to be an adequate primary management for CGIN, and the excision margin status of the LLETZ specimen did appear to be a prognostic factor for residual disease.

Distribution of Epithelial-Specific Antigen in Uterine Cervix with Endocervical Glandular Dysplasia

The natural history of cervical adenocarcinoma is largely unknown, and whether endocervical glandular dysplasia is a precursor lesion to malignant tumor formation is a controversial issue. The aim of this study was to clarify the relationship of endocervical glandular dysplasia to the uterine endocervical adenocarcinoma. Thirty-one cases of glandular abnormalities of the uterine cervix were identified histologically from January 1984 to April 1992. These included 11 cases of endocervical glandular dysplasia, 5 cases of adenocarcinomain situ,4 cases of microinvasive adenocarcinoma, and 11 cases of invasive adenocarcinoma. The immunohistochemical localization of epithelial-specific antigen (ESA) was examined in these cases of endocervical glandular dysplasia and related lesions, and was compared to 10 normal endocervical specimens. ESA immunoreactivities were usually present in the basolateral membrane of endocervical cells in the normal endocervix. However, the expressions of ESA were increasing from the basolateral membrane to the diffuse cytoplasmic membrane, along with the malignant transformation of the endocervical cells, in 6 of 11 cases with glandular dysplasia, in 9 of 9 cases within situand microinvasive adenocarcinomas, and in 11 of 11 cases with invasive adenocarcinomas. This finding indicates that ESA is a useful marker for endocervical glandular dysplasia and related lesions with malignant transformation, and suggests that endocervical glandular dysplasia may be a precursor lesion to uterine cervical adenocarcinomas.

Correlation of numerical chromosome 11 and 17 imbalance with metastasis of primary breast cancer to lymph nodes.

Abnormalities of chromosomes 11 and 17 have been widely reported in invasive carcinoma of the breast. Interphase cytogenetics using pericentromeric repeat probes allows the evaluation of numerical chromosomal aberrations in tumour cell populations. We have developed a method for interphase cytogenetics on fine needle aspirates taken from breast tumours and have applied it to the analysis of chromosomes 11 and 17 in 49 cases of invasive adenocarcinoma. Frequency distributions of signal number were generated for each case and no correlation was found between modal signal number and tumour size at presentation, nodal status or tumour differentiation. In 14 cases, two copies of each of chromosomes 11 and 17 were present, and in 14, the number of chromosomes 11 and 17 were equal but abnormal. In 14 cases, the chromosome 11 number was greater than chromosome 17 and in 7 cases, the chromosome 17 number was greater than chromosome 11. Chromosome inequality correlated with the presence of lymph node metastases or disseminated disease at presentation and the absence of in situ carcinoma. There was no relationship with the presence of vascular invasion. These data suggest that numerical chromosome 11 and 17 imbalance may indicate the ability of breast cancers to metastasize rather than invade vessels. The pattern of numerical chromosome abnormality described may define a subgroup of tumours with a greater tendency for metastasis.

Lipofuscin pigmentation (so-called "melanosis") of the prostate.

Although intraepithelial pigment in the prostate gland has been termed melanosis, the nature of the pigment is not entirely clear, and many pathologists are not aware of its existence. We examined 863 hematoxylin and eosin (H + E) stained slides from 150 surgical specimens of prostate (69 needle biopsies, 66 transurethral resections, 14 radical prostatectomies, and 1 suprapubic prostatectomy) from 149 patients (age range, 47 to 90 years; mean 70 years) in an effort to characterize this pigment. The 1-3 microns in diameter, predominantly subnuclear, yellow-brown to gray-brown granules with a dark blue rim (by H + E) stained positively with Fontana-Masson, periodic acid-Schiff with diastase, Congo red, luxol fast blue, and oil-red-O and exhibited yellow autofluorescence consistent with lipofuscin. H + E stained slides revealed pigment in the benign epithelium in 86 of 150 cases (57%), within stromal macrophages in eight cases, and in atypical epithelium in two cases of high-grade prostatic intraepithelial neoplasia. Ten cases of invasive adenocarcinoma without recognizable pigment in H + E stained sections were stained by the Fontana-Masson technique, and pigment was identified in malignant epithelium in three of these cases. Ultrastructural examination of intraepithelial pigment in KII-fixed tissue from three radical prostatectomy specimens demonstrated the typical appearance of lipofuscin. Although intraepithelial pigment in prostatic biopsy or resection specimens is usually considered characteristic of seminal vesicle epithelium, our study demonstrates that lipofuscin is commonly present in epithelial cells of benign prostatic hyperplasia and less frequently in those of prostatic intraepithelial neoplasia and adenocarcinoma. The recognition of this pigment is important in preventing diagnostic confusion with seminal vesicle epithelium and with melanocytic lesions.

Prostatic Invasive Adenocarcinoma: Effect of Combination Endocrine Therapy (LHRH Agonist and Flutamide) on the Expression and Location of Proliferating Cell Nuclear Antigen (PCNA)

Expression and location of Proliferating Cell Nuclear Antigen immunostaining in epithelial nuclei were assessed on histological sections from 32 cases of invasive adenocarcinoma of the prostate gland: 20 untreated and 12 treated with combination endocrine therapy or CET. The PCNA-positive nuclei showed homogeneous or granular types of staining or a mixture of both, and a gradation in the intensity of staining. Nuclei with homogeneous patterns appeared darker brown than the lighter granular and mixed patterns. Darker nuclei were more frequently noted, mainly among the epithelial cells adjacent to the stroma, in the untreated cases. In contrast, nuclei with pyknotic chromatin, unstained and corresponding to apoptotic bodies, were more frequently seen in the treated patients. For the untreated invasive adenocarcinomas, the mean proportion of PCNA-stained epithelial nuclei in the 10 cases with an acinar pattern (small and large) was 8.86% (SE 0.23%). The mean value in the 5 cases with a cribriform pattern was 11.76% (SE 0.52%), that is, greater than in the acinar pattern, and decreased from the nuclei in the basal position, or adjacent to the stroma, toward the lumen: 14.40% (SE 0.61%) in the basal position, 11.84% (SE 1.30%) in the intermediate and 9.26% (SE 0.66%) in the lumenal. In the 5 cases with a solid/trabecular pattern, the proportion of PCNA-positive nuclei was 15.74% (SE 2.30%), that is, higher than in all the other patterns, and decreased from the cell layer adjacent to the stroma (17.60%, SE 2.92%) toward the other layers (13.88%, SE 1.71%).(ABSTRACT TRUNCATED AT 250 WORDS)

Relationships between c-Ki-ras Mutations, HPV Types, and Prognostic Indicators in Invasive Endocervical Adenocarcinomas

Objective. Ras gene mutations have been identified in squamous cell carcinomas of the cervix and are associated with advanced-stage tumors. The current study was designed to determine the prevalence of c-Ki-ras mutations in invasive endocervical adenocarcinomas and associations between mutations, HPV status, and known prognostic indicators. Methods. Paraffin-embedded tissue samples from 32 cases of invasive adenocarcinoma of the endocervix were analyzed for the presence of c-Ki-ras mutations by amplification of DNA extracted from paraffin blocks using the polymerase chain reaction (PCR) and subsequent restriction enzyme digestion of the amplimers. This method allows detection of aspartic acid mutations of the c-Ki-ras gene at codons 12 and 13. Samples were also analyzed for the presence of HPV DNA using PCR and either "consensus sequence" primers or "type-specific" primers for HPVs 16 and 18. Patients' charts were reviewed for clinical information and correlations between clinical outcome and known prognostic indicators and the presence of c-Ki-ras mutations was assessed. Results. c-Ki-ras codon 12 mutations were detected in only 4 of the 32 cases (12.5%) of invasive endocervical adenocarcinomas examined. No codon 13 mutations were detected. Fifty-nine percent of the cases were HPV 16 or HPV 18 DNA positive and only 1 of the 4 cases with a codon 12 mutation was associated with HPV 16 or HPV 18. There were no significant associations between stage or grade of tumor or outcome and the presence of codon 12 mutations. However, using the observed survival as computed by the Berkson Gage Method at 12 months, the estimated median survival for patients with c-Ki-ras codon 12 mutations is 29 months compared to 68 months for patients lacking codon 12 mutations. Conclusions. These results indicate that c-Ki-ras codons 12 and 13 mutations are uncommon in invasive endocervical adenocarcinomas.

Histologic grade, clinical stage and DNA ploidy as prognostic indicators in prostatic carcinoma

53 cases of invasive adenocarcinoma of the prostate were retrieved from the files of CGMC over a two year period from 1985 -1986.Of these cases, 7 were considered to contain material unsatisfactory for DNA ploidy analysis by flow cytometry.The remaining 4 6 cases of formalin- fixed, paraffin-embedded tissues, were processed by a modified Hedley technique, stained with propidium iodide and analysed on a Coulter R Epics ProfileII flow cytometer with a 488nm air-cooled laser.A minimum nuclear count in each case of 5000 nuclei in the gated histogram of FL3 V FL3Peak was required for the assay to be considered as satisfactory.Mean % half-peak C.V. for the major peaks was 5.26.Median % half-peak C.V. was 5.66. of the 46 cases, 39 were DNA diploid, 2 aneuploid, and 5 tetraploid (major peak at 4n with >15% of total nuclei counted).There was no correlation between histologic grade, tumour stage and DNA ploidy.DNA ploidy showed no significance as an independent or multivariate prognostic indicator.

The prognostic significance of S-phase fraction analysis in carcinoma of the prostate

46 cases of invasive adenocarcinoma of the prostate diagnosed between 1985 and 1986 were retrieved from the files of CGMC. These cases were reviewed by on of us (HP) and were all graded according to Gleason. Seven other cases were rejected as having tissue unsatisfactory for processing by flow cytometry. The 46 cases were formed into a single nuclear suspension using a modified Hedley technique and the propidium iodide nuclear staining was measured using a Coulter^R Profile II flow cytometer with a 488nm air-cooled Argon laser. Each case had a minimum count of 5000 nuclei in the gated histogram FL3 V FL3Peak. The DNA histograms were assessed for DNA ploidy and S-phase fraction using the Multicycle^R software on an IBM-compatible 386 computer with a maths coprocessor. In all cases a background fitting algorithm for sliced nuclei was applied. Of the 46 cases, 39 were DNA diploid, 2 aneuploid and 5 tetraploid. Mean s-phase fraction for the diploid cases was 13.0% with a median of 10.2%. Sixteen of the 3 9 diploid cases had SPF > 13.0% and 19 cases > 10.2%. For the aneuploid cases the mean and median were the same with 20.8%. For the tetraploid cases the mean SPF was 8.2% with a median of 14.4%.