Expression Of Biological Markers Research Articles

Clear cell adenocarcinoma of the endometrium is a biologically distinct entity from endometrioid adenocarcinoma

Clear cell adenocarcinoma (CCA) of the endometrium has a poor prognosis, although the biologic features of this rare tumor are not clear. In this study, we analyzed the expression of biologic markers relating to carcinogenesis, tumor growth, and progression. Thirteen cases of CCA were compared with cases of endometrioid adenocarcinoma (EMA) of the endometrium. Immunohistochemical staining for p53; Ki-67; cyclins A, D1, and E; E-cadherin; progesterone receptor (PR)-A and PR-B; P-glycoprotein; MLH1; and MSH2 was performed. Labeling indices of p53, Ki-67, and cyclins A, D1, and E in CCA were 46.4 +/- 24.3%, 52.1 +/- 20.5%, 37.9 +/- 21.4%, 12.3 +/- 27.9%, and 8.2 +/- 22.9%, respectively. E-cadherin was expressed in only 1 case (7.7%) of CCA, as compared to 39 cases (61.0%) of EMA. No CCAs were positive for PR-A and PR-B. P-glycoprotein was detected in seven cases (53.8%). Loss of either MLH1 or MSH2 expression occurred in eight cases (61.5%). High-level expression of p53, cyclin A, and P-glycoprotein, and low-level or no expression of cyclin E, E-cadherin, PR-A, and PR-B was observed in CCA compared with EMA. The mechanism of cell-cycle regulation in endometrial CCA is different from that in EMA and may influence its malignant potential. Endometrial CCA is a distinct entity from EMA.

Frequent overexpression of epidermal growth factor receptor (EGFR) in mammary high grade ductal carcinomas with myoepithelial differentiation

Aims: To evaluate the expression of common biological markers and the epidermal growth factor receptor (EGFR) in mammary high grade ductal carcinomas with myoepithelial differentiation (DCMDs). Materials/methods: Thirty DCMDs were...

Male breast cancer: Clinico-pathological study with evaluation of biologic markers

777 Background: In contrast with female breast cancer, there is limited data on the expression of biologic markers and significance of clinicopathologic features in male breast cancer. Methods: We reviewed pathology and clinical data for 33 evaluable patients with MBC from 1981–1999. Immunohistochemical staining for ER, PR, AR, and cyclin D1 was performed. Results: Mean patient age was 61 years (range 35–80). 70% were Caucasian, 18% Hispanic, 12% African-American. 27% had a family history of breast/ovarian cancer in first-degree relatives. Patients presented as follows: with gynecomastia (33%), with a painless mass (33%), with nipple inversion/discharge (27%). Mean tumor size was 1.7 cm (range 0.1– 5.0 cm). The tumor was retroareolar/central in 67% patients. 88% had invasive ductal carcinoma while 12% had intracystic papillary DCIS. Tumor grade was as follows: high-grade (15), intermediate-grade (17), and low-grade (1). 40% had lymphovascular invasion, 15% presented with dermal involvement, and 12% had ni...

Male breast cancer: Clinico-pathological study with evaluation of biologic markers

777 Background: In contrast with female breast cancer, there is limited data on the expression of biologic markers and significance of clinicopathologic features in male breast cancer. Methods: We reviewed pathology and clinical data for 33 evaluable patients with MBC from 1981–1999. Immunohistochemical staining for ER, PR, AR, and cyclin D1 was performed. Results: Mean patient age was 61 years (range 35–80). 70% were Caucasian, 18% Hispanic, 12% African-American. 27% had a family history of breast/ovarian cancer in first-degree relatives. Patients presented as follows: with gynecomastia (33%), with a painless mass (33%), with nipple inversion/discharge (27%). Mean tumor size was 1.7 cm (range 0.1– 5.0 cm). The tumor was retroareolar/central in 67% patients. 88% had invasive ductal carcinoma while 12% had intracystic papillary DCIS. Tumor grade was as follows: high-grade (15), intermediate-grade (17), and low-grade (1). 40% had lymphovascular invasion, 15% presented with dermal involvement, and 12% had ni...

Dépression et facteurs protéiques de l'adaptation cellulaire : implication des traitements antidépresseurs

Mood disorders remain a puzzling clinical entity in psychiatry, not only at the symptomatic level (maniac versus depressive episodes, unipolar versus bipolar forms…) but also at the therapeutic level. Indeed, despite medications, the treatment responsiveness is rarely above 70%, occurs within the following weeks after initiation and is often not maintained on a half- to 1 year follow-up. The pioneering works of Axelrod’s and Brodie’s groups in the early 60s enlightening the ability of antidepressants to acutely increase central monoaminergic (serotoninergic or 5-HT, and noradrenergic or NAdr) activities have introduced the notion that these therapeutics affect the neuronal physiology. Since then, the affective disorders have been conceptualized as a biochemical disease corresponding to the mirror image of this antidepressant drug effectiveness. However, such understanding of the pathology minimizes or even denies the importance of some pharmacologically-induced events, e.g. the hypersensitization of postsynaptic 5-HT 1 A receptors combined with the desensitization of 5-HT autoreceptors (in particular of the 5-HT 1A subtype) which both occur once symptoms alleviate. This has likely weakened the dogma that treatments affect solely membrane-bound monoaminergic receptors. In addition, the use of drug combinations for fastening the clinical response (association with β-adrenergic drugs, anticonvulsants or thymo-regulators) have pointed out to speculate that the clinical drug effectiveness might be underlined by a sustained suractivation of the intracellular cascades triggered by the central monoaminergic systems (i.e. adenylate cyclase by cyclic adenosine monophosphate [cAMP] for the β-adrenergic, 5-HT 1A and 5-HT 1B/D receptor subtypes and calcium-dependent phospholipase and protein kinases by inositol triphosphate [IP 3] for the 5-HT 1C and 5-HT 2 receptor subtypes via heterotrimeric G protein-coupled receptors; Fig. 1). Accordingly, in the last decade, a new pathophysiological concept of affective disorders has thus emerged which integrates preferentially the molecular and cellular changes that supervene in response to an over-stimulation of these cascades. Most groups have focused their attention to the ability of antidepressants to enhance the expression and activation of a few transcription factors exemplified by CREB (AMPc response element binding protein) and neurotrophic factors such as nervous growth factor (NGF) and brain derived neurotrophic factor (BDNF) in the brain. Here, a review of the literature is synthesized on the topic that underlies the structural adaptations observed in the brain of depressed subjects and animals exposed to antidepressants. In fact, the hypothesis for a molecular vulnerability of mood disorders, a kind of neuronal memory for depression, has emerged in the early 90s when expression (identified by the messenger ribonucleic acid; mRNA) of various biological markers associated with depression [corticotrophin releasing hormone (CRH), tyrosine hydroxylase (TH), gluco-, mineralocorticoid and adrenergic receptors] was found to increase in parallel with the clinical response following a chronic antidepressant medication. Indeed, occurring with a different delay to the well-known mobilization of serotoninergic and noradrenergic receptors located at the plasma membrane level (that occur rapidly after initiation), the clinical response to antidepressant drugs has coincided with the long-term recruitment of various nuclear transcription factors, the mostly documented one being today CREB, and of various neurotrophic factors exemplified by BDNF. This recent hypothesis based on enhanced levels of both transcription and subsequent neurotrophic factors is nowadays getting more attractive due to specific documentation by both animal and human reports. However, whether quantifying expression (assessed by the mean of the mRNA levels) or protein activity of these factors is representative of the biological responsiveness to antidepressants still remains controversial since both hypotheses have now found their own defenders and detractors.

D�pression et facteurs prot�iques de l'adaptation cellulaireB: implication des traitements antid�presseurs

Les adaptations réceptorielles observées au cours du traitement de la dépression ne prédisent pas l'ensemble des ajustements biologiques opérés par la thérapeutique. L'objectif de ce travail est d'apporter les informations les plus récentes dans le domaine qui soient propices à une meilleure compréhension biologique de cette pathologie et de ses thérapeutiques. On sait depuis peu que les antidépresseurs modulent l'expression et l'activation de certains facteurs de transcription comme le CREB (AMPc response element binding protein) et de facteurs neurotrophiques comme le BDNF (brain derived neurotrophic factor) dans le cerveau. L'observation que certaines manifestations dépressives peuvent être induites (ou à l'inverse supprimées) par la réduction (ou à l'inverse l'augmentation) de ces protéines nucléaires suggère ainsi l'existence de véritables « mémoires biologiques » du processus dépressif par épuisement de ces facteurs et ouvre des perspectives nouvelles dans la thérapeutique de la dépression. Mood disorders remain a puzzling clinical entity in psychiatry, not only at the symptomatic level (maniac versus depressive episodes, unipolar versus bipolar forms…) but also at the therapeutic level. Indeed, despite medications, the treatment responsiveness is rarely above 70%, occurs within the following weeks after initiation and is often not maintained on a half- to 1 year follow-up. The pioneering works of Axelrod’s and Brodie’s groups in the early 60s enlightening the ability of antidepressants to acutely increase central monoaminergic (serotoninergic or 5-HT, and noradrenergic or NAdr) activities have introduced the notion that these therapeutics affect the neuronal physiology. Since then, the affective disorders have been conceptualized as a biochemical disease corresponding to the mirror image of this antidepressant drug effectiveness. However, such understanding of the pathology minimizes or even denies the importance of some pharmacologically-induced events, e.g. the hypersensitization of postsynaptic 5-HT1A receptors combined with the desensitization of 5-HT autoreceptors (in particular of the 5-HT1A subtype) which both occur once symptoms alleviate. This has likely weakened the dogma that treatments affect solely membrane-bound monoaminergic receptors. In addition, the use of drug combinations for fastening the clinical response (association with β-adrenergic drugs, anticonvulsants or thymo-regulators) have pointed out to speculate that the clinical drug effectiveness might be underlined by a sustained suractivation of the intracellular cascades triggered by the central monoaminergic systems (i.e. adenylate cyclase by cyclic adenosine monophosphate [cAMP] for the β-adrenergic, 5-HT1A and 5-HT1B/D receptor subtypes and calcium-dependent phospholipase and protein kinases by inositol triphosphate [IP3] for the 5-HT1C and 5-HT2 receptor subtypes via heterotrimeric G protein-coupled receptors; Fig. 1). Accordingly, in the last decade, a new pathophysiological concept of affective disorders has thus emerged which integrates preferentially the molecular and cellular changes that supervene in response to an over-stimulation of these cascades. Most groups have focused their attention to the ability of antidepressants to enhance the expression and activation of a few transcription factors exemplified by CREB (AMPc response element binding protein) and neurotrophic factors such as nervous growth factor (NGF) and brain derived neurotrophic factor (BDNF) in the brain. Here, a review of the literature is synthesized on the topic that underlies the structural adaptations observed in the brain of depressed subjects and animals exposed to antidepressants. In fact, the hypothesis for a molecular vulnerability of mood disorders, a kind of neuronal memory for depression, has emerged in the early 90s when expression (identified by the messenger ribonucleic acid; mRNA) of various biological markers associated with depression [corticotrophin releasing hormone (CRH), tyrosine hydroxylase (TH), gluco-, mineralocorticoid and adrenergic receptors] was found to increase in parallel with the clinical response following a chronic antidepressant medication. Indeed, occurring with a different delay to the well-known mobilization of serotoninergic and noradrenergic receptors located at the plasma membrane level (that occur rapidly after initiation), the clinical response to antidepressant drugs has coincided with the long-term recruitment of various nuclear transcription factors, the mostly documented one being today CREB, and of various neurotrophic factors exemplified by BDNF. This recent hypothesis based on enhanced levels of both transcription and subsequent neurotrophic factors is nowadays getting more attractive due to specific documentation by both animal and human reports. However, whether quantifying expression (assessed by the mean of the mRNA levels) or protein activity of these factors is representative of the biological responsiveness to antidepressants still remains controversial since both hypotheses have now found their own defenders and detractors.

Prognostic factors in soft tissue sarcomaTissue microarray for immunostaining, the importance of whole-tumor sections and time-dependence

In adult soft tissue sarcoma (STS) of the extremities and trunk wall, improved prognostic factors are needed to identify patients at high-risk for metastasis. Various factors are included in the many prognostic systems currently in use and the prognostic value of immunohistochemical (IHC) expression of biological markers is unclear. The tissue-preserving, high throughput tissue microarray (TMA) technique for analysis of immunohistochemical expression of biological markers was validated for Ki-67, and was found to yield results comparable to conventional staining methods. TMA was used to study the IHC expression of multiple markers (Ki-67, p53, cyclin A, bcl-2, beta-catenin, CD44, and Pgp) in 218 malignant fibrous histiocytomas (MFH) and in 140 mixed STS. In the MFH series, tumor size and Ki-67, as the only IHC marker, provided independent prognostic information. In the mixed STS series whole-tumor sections were used and TMA was performed in the peripheral tumor growth zone. Whole-tumor sections facilitated assessment of the strong independent prognostic factors for metastasis vascular invasion, hazard ratio (HR) 3.5, tumor necrosis (HR 2.8), and tumor growth pattern (HR 3.2), and the latter also correlated with local recurrence (LR). In comparison, histological malignancy grade, tumor size, and depth were not of independent prognostic value. When TMA was performed from the peripheral tumor growth zone, the IHC expression of Ki-67 (HR 1.9), beta-catenin (HR 2.7), CD44 (HR 2.1) and Pgp (HR 2.4) were independent prognostic factors. Finally, prognostic factors were found to be time-dependent, and most had lost their prognostic value after 2 years, whereas LR was a strong prognostic factor for metastasis whenever it occurred.

Biological markers in pT1 and pT2 ovarian cancer with lymph node metastases

Objective A relatively high incidence of pelvic and paraaortic lymph node metastases is found in patients with pT1 and pT2 ovarian cancer. This paper investigates the clinicomorphological parameters and the expression of various biological markers in these tumors in order to define possible risk factors for lymphatic dissemination. Methods In a retrospective study we identified 51 patients with pT1 and pT2 ovarian cancer. All patients underwent total hysterectomy, bilateral salpingo-oophorectomy, omentectomy, and systemic pelvic ± paraaortal lymphadenectomy. The incidence of lymph node metastases in these patients and the clinicomorphological parameters of their tumors were examined. Immunohistochemistry was used to determine the expression levels of the cell proliferation marker Ki-67, the cell adhesion molecules CD44s and CD44v6, and the oncoprotein HER2/neu of the tumors and their respective lymph node metastases. Results Lymph node involvement was found in 5 of 26 patients with pT1 ovarian cancer and in 6 of 25 patients with pT2 ovarian cancer. Serous adenocarcinoma was associated with a significantly higher incidence of lymph node metastases than other histological types (χ 2 = 4.7, P = 0.03). No correlation was found between tumor grade and the lymph node status. High Ki-67 expression was significantly correlated with spread to the lymph nodes (χ 2 = 4.2, P = 0.04), whereas expression of CD44s, CD44v6, and HER2/neu was not related to the lymph node status. Survival analyses showed no difference in disease-free and overall survival in patients with lymph node metastases compared to those without lymph node metastases. No association was seen among histological type, tumor grade, and immunohistochemically detected Ki-67, CD44s, CD44v6, and HER2/neu expression on the one hand and disease-free and overall survival on the other hand. Conclusions Our data suggest that in early stage ovarian cancer the serous histological type and tumors showing a high Ki-67 expression carry a high risk of lymph node metastases. With respect to prognosis our data showed a minor role for Ki-67, CD44s, CD44v6, and HER2/neu expression and the occurrence of lymph node metastases in pT1 and pT2 ovarian cancer.

Clinicopathologic analysis of pancreatic adenocarcinoma in African Americans and Caucasians.

African Americans have a higher incidence of pancreatic adenocarcinoma than do Caucasians for unknown reasons. Whether other clinicopathologic differences exist between these two groups is not known. This study was undertaken to compare the clinical, pathologic, and biologic findings for a group of patients with a histologic diagnosis of pancreatic ductal adenocarcinoma of the usual type in a single institution. We studied 410 patients (166 African Americans and 244 Caucasians) with a histologic diagnosis of pancreatic ductal adenocarcinoma of the usual type and analyzed (a) the clinicopathologic characteristics of the tumors, (b) the immunohistochemical expression of biomarkers implicated in pancreatic carcinogenesis (Fas, Fas ligand, HER2, p21/waf-1, p27, and p53), and (c) the presence and types of K- mutations at codon 12 as determined by polymerase chain reaction-mediated amplification. All elements of data were not available for all patients. African Americans had significantly higher rates of K-ras mutations to valine than did Caucasians (58% versus 22%, respectively; p = 0.015) and were less likely to have received chemotherapy (45% versus 70%, respectively; p = 0.001) or radiation therapy (34% versus 57%, respectively; p = 0.003). African Americans also had more frequent positive surgical margins than did Caucasians (56% versus 25%, respectively; p = 0.001), although mean tumor size was similar between the two groups (African Americans, 3.4 cm; Caucasians, 3.5 cm). Other clinicopathologic variables were similar between the two groups, including median survival (African Americans, 8.5 months; Caucasians, 10.1 months), 5-year survival (African Americans, 3%; Caucasians, 6%), and stage at presentation. Differences in biomarker immunoreactivity included less frequent Fas expression (4% versus 24%, respectively; p = 0.048) and a trend toward more frequent strong HER2 expression (39% versus 18%, respectively p = 0.11) in African Americans than in Caucasians. Although African American and Caucasian patients had similar survival rates associated with usual type pancreatic ductal adenocarcinoma, there were differences in management and expression of biologic markers between these two groups. African Americans were significantly less likely to receive radiation therapy or chemotherapy than were Caucasians, which may assume more importance as treatment improves. At the molecular level, African Americans had more frequent K- mutations to valine than did Caucasians.

Perineural invasion in adenoid cystic carcinoma: Its causation/promotion by brain-derived neurotrophic factor

Adenoid cystic carcinoma (ACC) is a common salivary gland neoplasm with a lengthy clinical course and often late local recurrences after surgical resection. This is accounted for histologically by its infiltrative capacity and distinct propensity for perineural invasion. The expression of biological markers may help explain the clinicopathologic course in ACCs. Brain-derived neurotrophic factor (BDNF) is a growth factor known to be involved in neurogenesis. The aim of this study is to elucidate the expression of BDNF in ACCs, which is currently unknown. Twenty-nine cases of primary ACCs of the head and neck were immunostained to analyze BDNF protein expression. Staining intensity was described as focal or diffuse and graded on a 3-tiered scale. The study group comprised 20 adult females (age 30 to 78) and 9 adult males (age 22 to 70). Sites of involvement included the parotid gland (6 cases), nasopharynx (5), maxilla (4), palate (3), trachea (3), submandibular gland (2), buccal mucosa (2), mandible (1), tongue (1), lacrimal gland (1), and temporal region (1). All tumors exhibited diffuse cytoplasmic staining; 11 cases were classified as 1+ intensity, 12 cases as 2+, and six cases as 3+. Based on the results presented here, BDNF is unformly expressed by ACC and may play a causative role in its predilection for perineural invasion. ACCs may display neurogenesis with high levels of BDNF expression in some tumors. Further studies are warranted to gain better understanding of this possible relationship. HUM PATHOL 33:933-936. Copyright 2002, Elsevier Science (USA). All rights reserved.

Calcium Dysregulation in the Aging Brain

The idea that age-related cognitive decline is associated with disruption of calcium (Ca2+) homeostasis has been investigated over the past two decades. Much of this work has focused on the hippocampus because hippocampal-dependent memory is age sensitive. It is now well established that Ca2+-dependent processes such as susceptibility to neurotoxicity, the afterhyperpolarization amplitude, induction of synaptic plasticity, and long-term potentiation and long-term depression are altered with age. Recent work has identified changes in Ca2+signaling pathways that may underlie the development of these biological markers of aging. This review considers recent findings concerning interactions between the various Ca2+-dependent processes, with special emphasis on the role of altered Ca2+ regulation and disruption of Ca2+ signaling pathways in mediating the expression of biological and behavioral markers of brain aging.

Comparison of potential biological markers cathepsin B, cathepsin L, stefin A and stefin B with urokinase and plasminogen activator inhibitor-1 and clinicopathological data of breast carcinoma patients

Cysteine, serine and metalloproteinases and their respective inhibitors are involved in tumor cell invasion and may have prognostic value for the outcome of malignant disease. The aim of the study was to compare the expression of new potential biological tumor markers, the lysosomal cysteine proteinases and their endogenous inhibitors, with that of the serine proteinases and their inhibitors in breast cancinoma and to relate their levels to the clinicopathological factors of the disease. Enzyme-linked immunosorbent assays (ELISAs) were used to measure cysteine cathepsin B (CatB) and cathepsin L (CatL) and their inhibitors, stefin A (StA) and stefin B (StB), together with urokinase (u-PA) and plasminogen activator inhibitor-1 (PAI-1), in 150 cytosols of primary invasive breast carcinoma. A good correlation was found between the levels of the two cysteine proteinases but only a moderate one between those of the cysteine and serine proteinases. u-PA and PAI-1 levels correlated positively with histological grade and negatively with estrogen receptor (ER) status. PAI-1 correlated with most clinicopathological factors that indicate the progression of the disease, while cathepsins and stefins were independent of these factors. In the total group of patients, high u-PA and PAI-1 and low StB levels correlated significantly with shorter disease-free survival (DFS), while CatB, CatL and StA did not. In lymph node negative patients, high CatB and CatL were also associated with shorter DFS, while u-PA remained the most significant of all these biological markers. In conclusion, this retrospective study showed u-PA to be of better prognostic relevance than the cysteine proteinases, though CatB and CatL were relevant for prognosis in lymph node negative breast cancer patients.

Apocrine ductal carcinoma in situ of the breast: Histologic classification and expression of biologic markers

Apocrine ductal carcinoma in situ (ADCIS) has been called a special type of ductal carcinoma in situ (DCIS) because the histologic grading is considered difficult using the classification schemes that have been proposed for common DCIS. However, ADCIS encompasses a spectrum of lesions with different morphologic aspects ranging from minimally atypical to overtly malignant. To define a classification scheme for ADCIS, 35 cases (22 pure and 13 associated with invasive carcinoma) were selected on the basis of conventional morphology on hematoxylin and eosin (H&E)-stained sections. Each case was assigned to 1 of 3 histologic grades (low, intermediate, and high) based on nuclear morphology and the presence of necrosis. In addition, the expression of hormone receptors p53, bcl-2, c-erbB-2, and Ki-67 was evaluated by immunohistochemistry, and the DNA ploidy was determined by image cytometry. Fifteen cases were classified as high histologic grade, 10 as low histologic grade, and the other 10 as intermediate grade. All but 4 cases, irrespective of grade, had the same hormonal immunophenotype: androgen receptor positivity (97.1%) and estrogen receptor and progesterone receptor negativity (94.3% and 97.1% respectively). Twenty-one cases (61.8%) showed p53 expression, and 47.1% of the cases were positive for c-erbB-2. The median positivity for Ki-67 was 5.2%. ADCIS has a unique morphologic and hormonal profile, distinct from common DCIS, deserving a specific classification. The proposed classification scheme allows for categorization of ADCIS according to the most important morphologic features already seen in common DCIS, ie, nuclear grade and necrosis. The expression of biologic markers other than hormonal receptors and bcl2 in ADCIS seems in general to be similar to that in common DCIS. Ki-67 and c-erbB-2 are expressed more frequently in intermediate and high histologic grade ADCIS. HUM PATHOL 32:487-493. Copyright © 2001 by W.B. Saunders Company

Immunohistochemical study during healing of free palatal mucosa grafts on plastic-embedded samples.

This immunohistochemical study describes changes in the histology and in the distribution of the basement membrane components laminin and collagen IV as well as of the cytokeratins (CK)1/2/10/11, CK5/6, CK13, CK14, CK17, CK19 during the take of free split mucosa (epithelial and connective tissue) transplants in humans up to 36 months post-operative. Histology showed a flattening of the epithelial layer within the first 2 weeks after grafting, followed initially by an increase (25-30 layers, week 6) and later on by a decrease of cell layers in the epithelium (15-20 layers, week 20). From that time onwards, clear stratification and reteridges as signs of differentiation were present. Up to day 14 of graft take, the linear staining patterns for laminin and collagen IV were interrupted, which was not observed at any later stage. During this early interval CK5/6, CK1/2/10/11, CK14 and CK17 were expressed in all epithelial layers. The reactions for CK5/6 and CK1/2/10/11 were less intensive. At 6 weeks, CK1/2/10/11 stained the intermediate and superficial layers, being consistent with findings after longer graft take. CK5/6 reacted in the basal and intermediate cell layers, and CK13, CK14 and CK17 reacted in all layers. In the following period up to week 20, CK5/6 were found in the parabasal cells of the intermediate cell layers and the basal cells. CK14 staining was confined to these cell layers too, but also showed some reaction in the superficial layers. CK13 and CK17 were still bound to all layers. At 7 months post-operative, CK5/6 and CK14 were seen in their typical localisation in the basal cell layer and the parabasal cells of the intermediate layers, CK17 was seen mainly in the intermediate layer and CK13 was seen in focal areas of all layers. Anti-CK19 reacted only with single basal and parabasal cells up to week 20. These results suggest that during healing of mucosal autografts there is a sequence of changes in the expression of cell biological differentiation markers that may involve an epithelio-connective tissue interaction before the typical patterns for the donor side were observed again on the gingiva or mucosa of the hard palate.

DNA Topoisomerase II-Alpha Immunoreactivity as a Marker of Tumor Aggressiveness in Invasive Breast Cancer

Objective: The nuclear enzyme DNA topoisomerase (topo) II breaks and rejoins DNA strands; its isoform topo IIα is associated with active cell proliferation of mammalian cells. The aim of this study was to examine the relationship between the expression of topo IIα and biological behavior markers in breast cancer. Methods: Formalin-fixed, paraffin-embedded tissue from 88 samples of infiltrating breast cancer was immunohistochemically stained for topo IIα. For each case, a topo IIα index was determined by image analysis. Similar indexes were available for Ki-67 protein, a known cell proliferation marker, and p53, bcl-2 and c-erbB-2 oncoproteins. Each case had been staged and graded and the patients had been followed up for a mean period of 61.62 months. Results: Elevated topo IIα immunopositivity (in >10% of malignant nuclei) was detected in 22 tumors, and this immunostatus was statistically associated with poor nuclear differentiation, absence of steroid hormone receptors, high Ki-67 immunoexpression, p53 protein accumulation and c-erbB-2 protein overexpression. Topo IIα expression was not linked with disease extent (stage or lymph node status). Neither proliferation marker (topo IIα or Ki-67) had any significant influence on the patients’ recurrence-free survival. Conclusion: From the above results, we conclude that topo IIα overexpression appears to be linked with cellular dedifferentiation and potentially aggressive tumor phenotype in invasive breast cancer.

Minimal breast cancer: evaluation of histology and biological marker expression.

Ninety-eight minimal breast cancers (MBCs) diagnosed between 1975 and 1990, and all originally considered to be invasive were found, on review, to form three groups: (a) 28 predominantly invasive carcinomas ≤10 mm (‘predominant invasive’); (b) 48 predominantly ductal carcinoma in situ (DCIS) lesions with definite foci of invasion each ≤10 mm (‘predominant DCIS’); and (c) 22 DCIS without evidence of invasion (‘pure DCIS’). Tumour histology and immunohistochemical expression of Ki-67, c-erbB2, p53, oestrogen receptor (ER), progesterone receptor (PR), and Bcl-2 were compared. The major finding was the contrasting features in the two invasive groups, with significant differences in their extent of invasion (P < 0.0001), tumour grade (P = 0.03), DCIS type (P = 0.008) and in marker expression. In the predominant invasive group, the infiltrative component was usually greater than 5 mm, low-grade and associated with well-differentiated DCIS. Expression of Ki-67, c-erbB2 and p53 was generally low, and that of ER, PR and Bcl-2 high. The predominant DCIS group in contrast had a much smaller, commonly high-grade, invasive component, usually with poorly differentiated DCIS and the reverse pattern of marker expression. Although not significant, survival of patients in the predominant invasive group was slightly better. These findings suggest that invasive MBCs should perhaps be treated as separate entities, in order to aid more appropriate selection of treatment. © 1999 Cancer Research Campaign

Processing Fine Needle Aspirates of Prostate Carcinomas for Standard Immunocytochemical Studies and In situ Apoptosis Detection

A method is described for making permanent histological sections of prostate carcinoma material obtained by fine needle aspiration (FNA) under ecography guidance. Smears made from prostate aspirates were used for diagnosis and from the same patient remaining aspirates were expelled into fixative filled microcentrifuge tube. Aspirates were pelleted and further processed to paraffin blocks. Permanent histological sections were obtained and each section was defined as satisfactory when it contained about 200 intact tumor cells. We have used these tumor sections and immunocytochemistry (ICC) procedures to study molecular biological marker expression. The technique described here has proven to be easy to use and offered a fast, reliable and cost-effective method to obtain suitable samples for standard ICC and in situ apoptosis detection from FNA prostate carcinoma. The method should be equally suitable for outpatient use on other tumors in which FNA and ICC or in situ apoptosis detection is likely to be helpful.

Angiogenesis and molecular biologic substaging in patients with stage I non—small cell lung cancer

Although complete surgical resection remains the primary treatment for localized stage I non-small cell lung cancer, the cancer recurrence rate is 25% to 40%. If one could identify, a subset of patients using molecular factors that contribute to tumour aggressiveness, one might improve prognosis in this group with additional treatment. High expression of angiogenesis factor viii has been associated with the presence of nodal metastases in breast cancer; here we examined its relation to survival with non-small lung cancer. We examined angiogenesis using immunohistochemistry on paraffin blocks of tumour from 275 consecutive patients with stage I non-small cell lung cancer, more than 68 months of follow-up, and a 64% 5-year survival. Angiogenesis was calculated from the microvessel number per ten 200 x microscope fields measured at the tumor periphery, in the center, and in the area of highest concentration. Measurements in the central area were inconsistent due to prominent necrosis. However, microvessel number recorded at the periphery and at the "hottest" are correlated well (r2 = 0.952; p = 0.001), and a significant survival advantage was noted for low-level expression at both areas (peripheral, p = 0.046; "hottest", p = 0.006). Multivariate survival analysis using angiogenesis, protooncogene erbB-2, tumor suppressor gene p53, and the proliferation marker KI-67 defined angiogenesis as the most significant prognostic factor in stage I lung cancer. This molecular biologic substaging system including angiogenesis for stage I non-small cell lung cancer is independent of routine histopathologic factors and revealed an additive adverse effect with expression of several biologic markers (5-year survival: no marker [n = 51] 81%, 1 marker [n = 82] 71%, 2 markers [n = 84] 54%, and 3 to 4 markers [n = 58] 49%; p = 0.0001).

Expression of several biologic markers as prognostic markers in non-small cell lung cancers

Expression of several biologic markers as prognostic markers in non-small cell lung cancers

1098 Expression of several biologic markers as prognostic factors in patients with non-small cell lung cancers

Despite modem diagnostic, staging, and therapeutic advances, esp. with molecular biologic techniques, the 5-year survival rate of all cases of lung cancer does not exceed 15%. With better understanding of tumor biology, one may improve survival through proper treatment. Here we present the clinical significance of several biologic markers as prognostic markers in patients with non-small cell lung cancers. The survival has correlated with the expressibility of proliferative cell nuclear antigen (PCNA), epidennal growth factor receptor (EGFR), p53 and/or blood group antigen A (BGAA) using immunohistochemistry in 46 cases patients with non-small cell lung cancers. The results were as follows: (1) The expression of BGAA was correlate with better survival in median survival and in 2-year survival and that of PCNA was correlated with worse survival in median survival and 2-year survival rate. (2) The expression of EGFR or p53 was not valuable to predict prognosis in non-small cell lung cancers. (3) With simultaneous applications of PCNA, EGFR and p53 immunostain, the patients with 2 or more negative expressions showed better prognosis than the patients with 2 or more positive expressions. In conclusion, it is suggested that the expression of blood group antigen may be a positive prognostic factor and that of PCNA may be a negative prognostic factor and also, the combination of expressions of PCNA, EGFR and p53 may be used as a negative prognostic factor.