Cyclin D1 protein expression in human thyroid gland and thyroid cancer.

Cell cycle progression is facilitated by cyclin dependent kinases (CDKs) that are activated by cyclins, including Cyclin D1 and inhibited by CDK inhibitors. Evidence of the involvement of cyclin gene alterations and over expression of various cyclins in human cancer is growing. The role of Cyclin D1 in malignant progression of papillary carcinomas of the thyroid has yet to be established. We therefore studied the expression of Cyclin D1 protein in thyroid carcinomas of young Kuwaiti patients (36 cases of conventional papillary thyroid carcinoma, 12 cases of its follicular variant, one case of tall cell thyroid carcinoma and one case of medullary carcinoma) using immunohistochemistry. In 23 patients (46%) circumscribed areas of cells were detected that showed a distinct to strong nuclear staining for immunoreactive Cyclin D1 whereas the remaining bulk of the carcinoma cells were negative or only showed a slight cytoplasmic staining. None of the tested clinical or path histological parameters showed a statistically significant correlation with the focal immunostaining. This does not rule out that the detected foci with positive nuclear Cyclin D1 immunostaining are areas where a progressive transformation to a more malignant phenotype occurs which eventually leading to lymph node and distant metastases.

Report of an International Network of Cancer Treatment and Research workshop on non-Hodgkin's lymphoma in developing countries.

The International Network of Cancer Treatment and Research (INCTR) recently organized a workshop on non-Hodgkin lymphomas (NHLs) in selected developing countries with the purpose of examining existing information relating to the pathology and management of these neoplasms, and identifying potential areas for research. This report provides a summary of the information presented and is focused primarily on the pathology of NHLs in children and adults. In most countries, the WHO classification of lymphomas was used and most participating centers included immunohistochemistry using a wide array of lymphoid antibodies as part of routine diagnosis. Some of the series had been reviewed by an external panel of experts. B-cell lymphomas accounted for 82-88% of all NHLs. The proportions of chronic lymphatic leukemia (4-6%), mantle cell lymphoma (MCL, 3-5%), and plasmacytoma (2-4%) were similar in the series presented. However, there was a significant variation in the proportion of follicular lymphoma (FL), which accounted for 15% and 11% in India and Kuwait, but less than 5% in Pakistan and Egypt. All of these frequencies are significantly lower than those reported in Western series. Diffuse large B-cell lymphoma accounted for about 35% of cases in India but for more 50% in other countries, but this difference was not accounted for by an increased incidence in a single lymphoma subtype in India, but rather an apparent paucity of several subtypes (such as mantle cell and marginal zone lymphomas (MZL)) in other series. There were relatively high frequencies of Burkitt lymphoma in Egypt (7%) and precursor T-cell lymphoblastic lymphoma in India (6-7%). Peripheral T-cell lymphomas (PTCLs) (not otherwise specified and angioimmunoblastic subtypes) accounted for 3-5% of NHLs, and extranodal lymphoma of T/NK cell type was rare (<1%). These differences in the relative proportions of NHL subtypes among developing countries and between developing countries and the rest of the world presumably arise from differences in environmental and genetic factors that influence lymphomagenesis and strongly suggest that more research in developing countries would provide valuable insights into the pathogenesis of lymphoid neoplasms.

Follicular variant of papillary thyroid carcinoma -- a cytological study.

The cytological diagnosis of classical papillary carcinoma is easily established based on the characteristic architectural and nuclear features. However, the follicular variant of papillary thyroid carcinoma(FVPTC) poses a diagnostic challenge. In this study we analysed the cytological features of 14 histopathologically proven cases of FVPTC. We inferred that a combination of architectural features such as follicles and syncytial clusters and nuclear features, viz grooves, pseudoinclusions and enlarged nuclei with fine chromatin, were helpful in establishing the diagnosis. It is hence suggested that based on the combination of the aforesaid features a diagnosis of FVPTC be offered whenever it is possible. This helps in patient management, obviating the need for a second surgical intervention.

Prevalence of endometrial proliferation in pipelle biopsies in tamoxifen-treated postmenopausal women with breast cancer in Kuwait.

OBJECTIVE: To determine the prevalence of pathologic changes in the endometrium of tamoxifen-treated asymptomatic postmenopausal patients with breast cancer. SUBJECTS AND METHODS: Fifty postmenopausal asymptomatic breast cancer patients with positive estrogen receptor status were treated with 20 mg of tamoxifen daily for a period of 5-60 months. The control group consisted of 30 asymptomatic postmenopausal breast cancer patients who were negative for estrogen receptor and therefore did not receive tamoxifen. Endometrial biopsies were performed using Pipelle endometrial suction curette at least 5 months after the study began. The endometrium was classified as atrophic (negative finding) and proliferative or hyperplastic (positive findings). The study and control groups were compared for demographic characteristics, risk factors for endometrial cancer, histological findings and the duration of tamoxifen treatment. RESULTS: A significantly greater prevalence of endometrial abnormalities existed among the tamoxifen-treated than control patients (76 vs. 33%, p < 0.001). The abnormal endometrial changes were further demarcated in both groups into proliferative (54 vs. 26.7%, p = 0.02) and hyperplastic (22 vs. 6.6%, p = NS). In the study group, 63.6% of hyperplastic endometrium was simple hyperplasia and 36.4% was complex/no atypia hyperplasia, while in the control group all the cases were simple hyperplasia. No endometrial cancer was detected in either group. In addition, there was a positive association between the duration of tamoxifen exposure (<1 year vs. >/=1 year) and the endometrial abnormalities (46.6 vs. 88.6%, p = 0.003; proliferative 57.1 vs. 74.1%, p = 0.015; hyperplastic 42.8 vs. 25.8%, p = NS). CONCLUSION: The adjuvant use of tamoxifen is associated with significant time-dependent abnormal endometrial changes among patients with cancer of the breast.

Clinicopathological features of extranodal lymphomas: Kuwait experience.

A total of 935 patients with extranodal non-Hodgkin lymphoma (NHL) diagnosed in the period between January 1985 and December 2000 in Kuwait Cancer Center, serving the whole population of Kuwait, were used to describe the clinicopathological and epidemiological features of extranodal lymphomas in Kuwait. Extranodal lymphomas accounted for 45% of all NHL observed during this time. All NHL cases from Kuwait Cancer registry were analyzed and pathologically reclassified using the latest WHO (2000) classification. The most common lymphoma observed was diffuse large B-cell lymphoma (58.60%) followed by Burkitt s lymphoma (BL) (3.80%). In the pediatric group, BL comprises more than two thirds of all patients (77.20%). The most common extranodal sites were stomach (19.70%) and skin (17.80%) in the adult group, large intestine (29.80%) and small intestine (19.30%) in the pediatric age group. The majority (73.40%) of adult extranodal lymphomas was in stage IE-IIE and had a very good prognosis. On the contrary, the majority of pediatric extranodal lymphomas were found to be in stage III and IV. Variations in treatment policies (single agent or combined chemotherapy, radiotherapy, combined modality treatment) adopted and changed during the time period of 16 years of this retrospective study were documented.

Clinical implications of urokinase and tissue type plasminogen activators and their inhibitor (PAI-1) in breast cancer tissue.

Cytosol of primary breast cancers from 217 women of predominantly Arab ethnicity were assayed for uPA, tPA, PAI-1 and a subset for ER, PR and pS2. Serum levels of CEA and CA153 were determined during follow-up. Only tPA correlated to nodal status and tumour grade, and PAI-1 to clinical stage. PAI-1 was related to uPA and both were inversely correlated with PR and pS2 (PAI-1 also to ER). Conversely tPA was directly correlated with ER, PR and pS2. Women with high tumour uPA and PAI-1, but not tPA, had shorter overall, and relapse-free, survival. Only nodal status and clinical stage were independent predictors in multivariate analysis. However, uPA and PAI-1 were more prognostically informative than ER or PR and their usefulness may extend to delineation of patients likely to respond to adjuvant therapy.

Immunohistochemical detection of p53 protein expression in breast cancer in young Kuwaiti women.

The mutation of the p53 gene is a common phenomenon in numerous human tumors including breast cancer. It leads to an accumulation of nonfunctioning p53 protein in the cell nuclei, which can be detected by immunohistochemical techniques. In breast cancer overexpression of mutated p53 protein has been correlated to a poor prognosis. Our study is an immunohistochemical analysis of p53 in 82 cases of breast cancer in young (< or = 30 years old) Kuwaiti women, correlating it with histopathological grade, lymph node status, estrogen (ER) and progesterone receptor (PgR) content, tumor cell proliferation (immunostaining for Ki-67) and expression of c-erbB-2 oncoprotein. p53 immunostaining was found in 47 (57.32%) of the carcinomas. 65% of them displayed positive immunostaining for c-erbB-2. 63.7% of tumors with p53 overexpression were aneuploid. 64.8% of the p53 positive tumors were node positive. 93.5% of the p53 immunopositive carcinomas were ER-negative, and in 95.7% of this subclass of patients no PgR could be detected. The vast majority of p53 positive carcinomas were grade III (76.6%), 21.3% were grade II and 2.1% grade I, but neither tumor grade or tumor size showed a correlation with p53 expression. A significant negative correlation between ER- and PgR-content (p = 0.006) and immunostaining for p53 was observed. Our study provided evidence that the association of negative hormone receptor status and positivity for p53 immunostaining points to a greater tumor aggressiveness.

Evaluation of prognostic factors in male breast cancer.

We conducted an analysis on 41 cases of male breast cancer (median age 54 y; range 25-82 y) in Kuwait. Most (51%) were stage II cancers with 65% arising in the left breast. There were 5 (12%) T1 tumours, 23 (56%) T2 tumours and 13 (32%) T3/T4 tumours. They were mostly (95%) infiltrating ductal carcinomas; 97% were grade 2 or 3. Axillary lymph node involvement was found in 69%. Estimated 5-year survival rates were 67% and 58% for overall and relapse free survival respectively. Favourable prognosis was associated with age below 50y, clinical stage I and II, small tumour size (T1, T2), low tumour grade and absence of nodal involvement or distant metastases; nodal status and grade were independent factors for relapse free survival in multivariate analysis. In 18 cases, an immunohistochemical study showed some degree of tumour antigen reaction for ER in 89% of cases, PR in 61%, pS2 in 44%, CathD in 72%, p53 in 56%, c-erbB-2 in 50%, Ki67 and PCNA in 100% and bcl-2 in 78%. There were significant associations between several of these factors but none influenced survival. Despite the high incidence of staining of ER, our data do not support the concept of an endocrine pathway that could be usefully antagonized with antioestrogens for therapeutic benefit, as in women.

Expression of growth hormone receptor in human liposarcomas and lipomas.

Our immunohistochemical results clearly demonstrated the occurrence of growth hormone receptors (GH-R) in the tumour cells of lipomas and liposarcomas. In liposarcomas staining intensity in the cytoplasm of tumour cells varied between weak and distinct but could not be correlated to the histological grade of the malignant tumours. These findings were corroborated to some extent by the RT-PCR results. RT-PCR analysis of human lipomas and liposarcomas revealed the amplified cDNA fragment of GH-R in 8 out of 12 lipomas but only in 3 out of 10 liposarcomas. The reduced number of GH-R positive tumours found with PCR may be explained by the extraction method of RNA from paraffin sections. An interesting finding was the distinct immunoreactivity of the endothelium of blood vessels in liposarcomas, which was especially pronounced in the newly forming capillaries. This points to an important role of GH-R in tumour angiogenesis which could significantly contribute to tumour growth in liposarcomas and may open the possibility for therapeutic intervention using antiangiogenic substances.

Immunoradiometric measurement of pS2 in breast cancer--correlation with steroid receptors and plasminogen activators.

pS2 was measured by radioimmunometric assay in tumour extracts from 197 breast cancer patients. Values ranged from 0 to 50 ng/mg protein (mean 9.6 and median 3 ng/mg). We found no correlation with age, menopausal status, nodal metastases, disease stage or tumour histology. There was, however, a linear relationship with both ER (p < 0.0001) (particularly nuclear ER) and PR (p < 0.0001) expression determined by enzyme immunoassay (ELISA), as well as a good correlation when high and low expressors were stratified on the basis of combined ER/PR expression using consensus cut-off points. Only 15% of ER - ve/PR - ve patients were classified as pS2 + ve compared with 83% of those who were ER + ve/PR + ve. pS2 was also directly correlated with high expression of tPA and inversely with uPA. Comparison with previous studies showed that the current ELISA method produced consistent results, in contrast to other methods, particularly those based on immunohistochemical detection. The close relationship between pS2 and both steroid receptors suggests that pS2 may be important in terms of defining hormone-responsive patients who are likely to benefit from endocrine therapy.

Measurement of serum N-acetyl beta glucosaminidase activity in patients with breast cancer.

N-acetyl-beta-glucosaminidase (NAG) activity measured in sera from 129 breast cancer patients was elevated (mean 18.2 units/l) compared with that in sera from 28 healthy women (11.6 units/l) (p=0.001). There was a weak correlation between NAG activity and carcinoembryonic antigen (CEA) and CA-153, but no relationship to age, menopausal status, node status, stage, histology of tumour or to steroid receptors. NAG, CEA and CA-153 were measured in periodic follow-up samples taken after surgery (up to 26 months) from 17 patients. NAG activity fluctuated within a narrow range, unlike CEA and CA-153. In 70% of cases the pattern was similar to at least one of the other markers, and was generally maintained at a higher level in patients who suffered relapse compared with those who remained disease-free up to the last follow-up, but was not significantly altered before relapse. The measurement of NAG activity is unlikely to be of value in predicting time or occurrence of relapse or of clinical utility in post-surgical therapy. Increased appearance in serum may aid metastasis by degrading the extracellular matrix and it may be better investigated as a predictor of progression from in situ to invasive and metastatic cancer.

Steroid receptor measurement in breast cancers: comparison between ligand binding and enzyme-immunoassay in cytosolic and nuclear extracts.

We have analysed cytoplasmic and nuclear extracts of breast-cancer tissue from a total of 799 patients, measuring both oestrogen and progesterone receptors (ER, PR) using either the ligand binding assay (LBA) or the enzyme immunoassay technique (EIA). Mean and median receptor levels were much lower than those widely reported by others. For ER, this may in part be a consequence of the younger median age of the patient group. The frequency of positivity, using consensus cut-off values for clinical evaluation, was also lower than that reported by the EORTC Receptor Study Group. Although the measurements comparing the 2 methods were statistically correlated in terms of positivity, based on the above criteria for clinical assessment, concordance was considered to be relatively poor, particularly for ER when assayed in the same samples by the 2 methods. In cytosolic but not nuclear extracts, the LBA method gave a higher median value for ER than the EIA (except in the group that had EIA values greater than 15 fmol/mg protein); for PR, median values were higher with EIA in both cell fractions. There was an excellent correlation between receptor amounts in cytosolic and nuclear extracts for both ER and PR using the EIA; this was significantly better than with LBA. We also observed a correlation between ER and PR in both cytosolic and nuclear fractions which was most pronounced when the analysis was done by EIA. The amounts of ER in the cytosolic fraction were also correlated with the those of PR in the nuclear fraction and ER in the nuclear fraction with PR in the cytosolic fraction, but only when the EIA method was used. We conclude that the EIA method appears to be more sensitive and gives biologically more reliable results. However, the disagreement between the methods may be due to legitimate recognition of altered forms of the receptor and may be of biological significance. Although the presence of receptor in the cytosolic fraction is artifactual, its measurement by EIA does parallel the amounts of nuclear receptor, which may be a more relevant biological parameter.

Subpopulations of stromal cells from long-term human bone marrow cultures: ontogeny of progenitor cells and expression of growth hormone receptors.

Long-term culture of bone marrow derived stromal colony forming cells (S-CFC) in matrix and nutrient defined agar medium resulted in stromal cell colonies that pass sequentially through three distinct morphological stages: firstly, aggregated loose syncytium of round to avoid cells (stage I), a second developmental stage of large branching colonies in which the cells become enlarged, elongated with cytoplasmic projections forming a loosely anastomized network with adjacent cells (stage II), and finally cells become dissociated, loosing their long, thin cytoplasmic filaments and breaking their contacts with one another, but remain large and retain a bi-polar nature (stage III). Cells were also grown in liquid medium in a culture microenvironment closely resembling conditions of haemopoiesis in vitro. Using a panel of well defined monoclonal antibodies reactive against the rat, rabbit and human growth hormone receptors, this study found immunochemical evidence of the presence and localization of binding sites of growth hormone (GH) in the cell membrane and extra-nuclear Golgi area of long-term bone marrow derived human stromal cells in liquid and semi-solid nutrient agar mediums. GH-receptor immunoreactivity was present in small proliferating progenitor cells, myofibroblast-like cells, large reticular fibroblast cells, adipocytes and endothelial cells. Only MAb known to be reactive against human tissue resulted in strong immunoreactivity. The expression of GH-receptors not only on small proliferating, but also on the well differentiated cells, indicates a role for growth hormone on non-progenitor cells. GH-receptor immunoreactivity on differentiating and/or differentiated cells suggests that GH is also necessary for, or has a trophic function in differentiation. We propose that direct GH action is necessary not only for differentiation of progenitor cells as implied by the dual effector hypothesis, but also their subsequent clonal expansion, differentiation and maintenance.

Papillary carcinoma of the thyroid with numerous spindle-shaped tumor cells in fine needle aspiration smears. A case report.

A papillary carcinoma of the thyroid characterized by numerous spindle-shaped tumor cells in fine needle aspiration (FNA) smears is reported. The oval nuclei of the spindle cells, arranged in monolayered sheets, occasionally showed "rhythmic," palisaded patterns. Comparative study of cytology and histology of this tumor revealed that the spindle cells observed in FNA smears originated in the epithelium covering the edematous neoplastic papillae.

Estrogen and progesterone receptor status in breast cancer in Kuwait female population.

The levels cytosol estrogen (ERc) and progesterone (PRc) receptors were determined in 315 primary breast cancers of female Arab patients aged 23-80 years. Most of breast cancers (78%) occurred in women aged 21-50 years, and only 22% were in women aged 51-80 years. Breast cancers containing ERc and PRc concentrations in the range 5-50 fmol/mg of cytosol protein (mg c.p.) were found with similar frequency in women aged under or over 50 years (53% for ERc and 43% for PRc, respectively). On the other hand, breast cancers with ERc values of > 50 and > 100 fmol/mg c.p. were twice as frequent in women aged over 50 years as in women aged under 50 years. The frequency of breast cancers with PRc level of over 50 fmol/mg c.p. in women aged over 50 years was only half that in those aged under 50 years. In breast cancers of Kuwait Arab women the higher values of ERc (> 100 fmol/mg c.p.) and PRc (> 50 fmol/mg c.p.) were less frequent than in other populations reported in literature. The low frequency of breast cancer in postmenopausal Kuwait women is associated with low proportions of tumors with higher ERc and PRc contents. In contrast to this, data from literature indicate that in the North Western European and American populations the postmenopausal incidence rise of breast cancers is associated with increased proportions of tumors with higher ERc and PRc levels.

Relations between epidermal growth factor receptor and oestrogen and progesterone receptors in breast cancers of premenopausal and postmenopausal patients in Kuwait.

The levels of cell membrane epidermal growth factor receptor EGFR and cytosol (c) and nuclear (n), oestrogen (E) and progesterone (P) receptors (R) were determined in 132 specimens of primary breast cancers. In the tumours of postmenopausal women an inverse significant correlation was demonstrated between the concentrations of EGFR vs. ERc, ERn, and PRc while no such correlation was noted in the tumours of premenopausal women. Premenopausal and postmenopausal EGFR positive tumours (> or = 10 fmol/mg membrane protein) could be regarded as homogenous with respect to the concentration of ER and PR whose mean values were low and without being significantly different. EGFR negative tumours were heterogeneous with respect to the ER and PR concentrations. Postmenopausal EGFR negative (< 10 fmol/mg membrane protein) tumours had evidently higher mean values of ER and PR than premenopausal EGFR negative tumours, but these differences were statistically significant for oestrogen receptors only. The levels of ER and PR of premenopausal EGFR negative tumours were approximated to the corresponding levels of EGFR positive tumours.