Metastasizing APUD cell tumours of the human gastrointestinal tract. Light microscopic and karyometric studies

A total of 11 metastasizing gastrointestinal APUD cell tumours from biopsy and autopsy files were reclassified according to Soga and Tazawa as well as to WHO Histologic Classification of Tumours. The much higher proportion of APUD cell tumours in autopsies (11 cases from 1000 autopsies in comparison with 22 cases from 22 000 biopsies) demonstrate that the majority of them will not be discovered during the patient's life. EC cell carcinoids (type A) predominate in both non-metastasizing and metastasizing gastrointestinal APUD cell tumours. Metastases from EC cell carcinoids occurred only in regional lymph nodes and in the liver. The APUD cell tumours originating in the pancreas represent the most frequently metastasizing gastrointestinal carcinoids. Besides in the liver and in regional lymph nodes metastases from pancreatic APUD cell tumours were seen in the skin, the brain and the skeleton. One case with two (pancreatic, bronchial) competitive primary APUD cell tumours and a skin metastasis was studied by means of automated cell image analysis. Cell populations of these three tumour sites were characterized by morphometric and densitometric nuclear parameters. It could be demonstrated that the skin metastasis consisted of a cell population, which occurred as a subpopulation in the primary tumour of the bronchus. The results of karyometric investigations supported the hypothesis that single components of tumours can metastasis selectively.     

Cellular DNA content and metastasis pattern in colorectal carcinomas

Summary Tissue samples from primary tumours and metastases obtained from surgical specimens and autopsies were investigated by flow cytometry for their DNA content. Of the 149 cases investigated, 124 autopsy cases of colorectal carcinoma were suitable for study. The pattern of metastatic spread in each indiviual case was analysed with reference to the autopsy records. A subgroup of euploid primary tumours was observed which should be considered separately with regard to its biological behavior, tumour location and extent of metastases. In these tumours, the liver was the first and final organ of haematogenic metastatic spread at an above-random frequency and a grossly metastatic liver with less than 15% of residual normal parenchyma was present at the same time. Furthermore, we observed stem-lines deviating in ploidy from the primary tumour in the metastases of nine cases. This indicates the chromosomal heterogeneity of colorectal carcinomas.     

Primary and secondary heart tumours in dogs and cats

Primary and secondary neoplasms of the canine and feline heart are uncommon. During a 2-year period, 83 dogs suffering from primary cardiac (n=11), extracardiac benign (n=6) or malignant (n=66) tumours and 30 cats with primary cardiac (n=1) or extracardiac (n=29) malignant tumours were examined. Echocardiography revealed four cases of primary cardiac neoplasms in dogs, but secondary heart tumours were not detected. After necropsy, tissue samples from the heart and tumours were examined histologically and immunohistochemically. In dogs, primary neoplasms included seven haemangiosarcomas, two chemodectomas, one rhabdomyosarcoma, and one neurofibrosarcoma. In 24 of 66 dogs examined, metastases of extracardiac neoplasms were found in the heart (15 carcinomas, six malignant lymphomas, three haemangiosarcomas). In cats, one case of primary haemangiosarcoma of the pericardium and five cases of secondary cardiac tumours (two malignant lymphomas, three carcinomas) occurred. Cardiac neoplasms in cats were not identified clinically but were detected by detailed gross sectioning of the heart (n=2) or histopathological examinations (n=3). This study showed an unexpectedly high number (36%) of dogs with cardiac metastases.     

Differential expression of cell cycle and apoptosis related proteins in colorectal mucosa, primary colon tumours, and liver metastases

AIMS: Tumour cell growth results from a disturbance in the balance between the rate of proliferation and cell death. In this study, proteins involved in the regulation of cell cycle arrest and apoptosis were studied as possible factors responsible for uncontrolled cell growth in colorectal cancer. METHODS: The expression of proteins involved in these processes was investigated in 48 metastases from patients with colorectal cancer and compared with eight normal colon mucosa samples and 14 primary tumours. Both primary tumours and metastases were obtained from eight patients. The expression of thymidylate synthase (TS), p53, retinoblastoma protein (Rb), Fas receptor, Fas ligand, bcl-2, mcl-1, bax, and bcl-x was measured using immunohistochemistry. Proliferation was determined by Ki67 staining, whereas apoptosis was assessed by M30 immunostaining, which recognises cleaved cytokeratin 18. RESULTS: In the limited number of cases in which paired comparisons were possible, the expression of TS and Ki67 was significantly higher in metastases than in the matched primary tumour samples (p = 0.014 and 0.016, respectively), whereas Rb expression was lower in metastases than in primary tumours (p = 0.024). Fas receptor expression was high in normal mucosa but absent in primary tumours and metastases, whereas the opposite was seen for p53. The expression of bax, mcl-1, and bcl-x in normal mucosa was more apical than that seen in malignant cells, where a more diffuse expression pattern was seen (p < 0.04). Apoptosis was more abundant in primary tumours than in metastases. CONCLUSIONS: These results demonstrate that proliferation and apoptosis are disturbed during colorectal cancer progression, and this is accompanied by loss of Rb and Fas expression, the accumulation of p53 and TS, and changes in the expression patterns of bax, mcl-1, and bcl-xl.     

Change of HER-2/neu status in a subset of distant metastases from breast carcinomas

It is assumed that HER-2/neu status remains consistent in breast carcinoma during metastatic spread, but in most previous studies primary tumours were compared with concurrent regional lymph node metastases. The present study investigated 31 breast carcinomas and their corresponding lymph node and distant metastases for HER-2/neu status by immunohistochemistry (HercepTest) and fluorescence in situ hybridization (FISH) (PathVysion). In 14 cases, serum HER-2/neu (SHER-2) was measured sequentially using the Bayer Immuno 1 HER-2/neu assay. Comparing HER-2/neu immunohistochemistry of primary tumours and distant metastases case by case, increased HER-2/neu expression was found in the distant metastases in 15 cases (48.4%), three (9.7%) of which showed an increase from score 0 to score 3+. In contrast, lymph node metastases showed the same HER-2/neu expression as the primary tumours, confirmed by FISH. Two cases, which showed HER-2/neu score 3+ and HER-2/neu amplification in the primary tumours, revealed increased SHER-2 levels above 50 ng/ml at the first measurement. Five of the 14 cases (36%) showed an increase of SHER-2 above 50 ng/ml towards the end of the patients' life. On the basis of these results, there is evidence that in a subset of breast carcinomas, HER-2/neu amplification and overexpression occur de novo in distant metastases at a late disease stage.     

Malignant renal rhabdoid tumour. Immunohistochemical and ultrastructural studies

In an effort to establish their possible histogenesis, three cases of renal rhabdoid tumour and their metastases were studied both by a battery of polyclonal and monoclonal antibodies using the avidin-biotin-peroxidase complex technique and by electron microscopy. Vimentin was demonstrated in renal rhabdoid tumour in two cases and in all metastatic deposits. Cytokeratin (39, 43 and 50 kD) was not demonstrable in the three renal rhabdoid tumours, but was strongly positive in all metastatic lesions in one case. Epithelial membrane antigen was present in one renal rhabdoid tumour and in pulmonary metastases in two cases. Ultrastructural study showed epithelial differentiation in all tumours: basal lamina and convergent tight junctions were demonstrated; intracytoplasmic intermediate filaments were present in all primary and metastatic tumours. Rhabdoid tumours thus exhibited heterogeneous immunophenotypic expression suggesting that they are derived from mesenchymal cells which are capable of differentiating into epithelial cells.     

Intratumoural cytogenetic heterogeneity of sporadic colorectal carcinomas suggests several pathways to liver metastasis

Much has been learned about the chromosomal abnormalities of colorectal carcinomas but the cytogenetic relationship between the neoplastic clones present in primary versus metastatic tumour samples remains unclear. We analyse the frequency of abnormalities for 47 chromosome regions using the interphase fluorescence in situ hybridization technique in a group of 48 tumours, including 24 primary colorectal tumours and 24 paired liver metastases. All tumours showed complex karyotypes with numerical/structural abnormalities for seven or more different chromosomes/chromosome regions both in the primary tumours and in their paired metastases. Chromosome 8 was the most frequently altered (22/24 primary tumours), consistently showing del(8p22) and/or gains/amplification of 8q24, followed by abnormalities of the entire chromosome 7 (21/24 primary tumours) and of chromosomes 17p and 20q (20/24 primary tumours). Simultaneous staining for multiple chromosome probes revealed the presence of two or more tumour cell clones in 23/24 cases (46/48 tumour samples). Interestingly, the liver metastases typically contained tumour cell clones similar to those found in the primary tumours, suggesting the absence of selective selection of specific tumour clones. Despite this, additional chromosomal abnormalities were detected in 23/24 metastatic tumours, which preferentially consisted of del(17p13) and gains/amplification of 11q13 and 20q13; moreover, compared to primary tumours, metastases showed an increased number of abnormalities of chromosomes 1p, 7q, 8q, 13q, and 18q, and new chromosomal abnormalities involving chromosomes 6, 10q23, 14q32, 15q22, and 19q13. Owing to the high frequency of numerical abnormalities of the entire chromosome 7 and loss and/or gain/amplification of specific regions of chromosome 8, eg del(8p22) and/or gains/amplification of 8q24 in primary colorectal tumours with associated metastases, it is suggested that their assessment at diagnosis could be of great clinical utility for the identification of colorectal cancer patients at higher risk of developing liver metastases. Copyright © 2010 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.     

Gastric schwannoma—a clinicopathological analysis of six cases

Six gastric schwannomas found among 150 mesenchymal tumours of the gastrointestinal tract were analysed clinically, histologically and immunohistochemically. These tumours occurred in the wall of stomach in middle-aged patients, five women and one man, and measured 2–9 cm in diameter. Follow-up from 3 to 24 years showed no recurrences or metastases. Histologically, all tumours were composed of spindle cells with vague nuclear palisading and variably myxoid stroma. One case showed a multinodular pattern of growth, reminiscent of plexiform neurofibroma. In all cases, there was a peripheral cuff-like B-lymphocyte infiltration with germinal centers in two cases. None of the tumours showed mitotic activity. Immunohistochemically, the tumour cells were positive for S-100 protein and focally for GFAP and CD57 (Leu 7). They were negative for desmin and actin, unlike true leiomyomas, and negative for CD34, unlike most gastrointestinal stromal tumours that were examined for comparison. Electronmicroscopy of three cases showed complex cell processes surrounded by prominent basement membranes, while myofilaments were not present. These cases show that schwannomas can be identified as rare, benign gastrointestinal tumours which probably arise from the gastrointestinal autonomic nervous system.     

Comparative genomic hybridisation as a supportive tool in diagnostic pathology

AIMS: Patients with multiple tumour localisations pose a particular problem to the pathologist when the traditional combination of clinical data, morphology, and immunohistochemistry does not provide conclusive evidence to differentiate between metastasis or second primary, or does not identify the primary location in cases of metastases and two primary tumours. Because this is crucial to decide on further treatment, molecular techniques are increasingly being used as ancillary tools. METHODS: The value of comparative genomic hybridisation (CGH) to differentiate between metastasis and second primary, or to identify the primary location in cases of metastases and two primary tumours was studied in seven patients. CGH is a cytogenetic technique that allows the analysis of genome wide amplifications, gains, and losses (deletions) in a tumour within a single experiment. The patterns of these chromosomal aberrations at the different tumour localisations were compared. RESULTS: In all seven cases, CGH patterns of gains and losses supported the differentiation between metastasis and second primary, or the identification of the primary location in cases of metastases and two primary tumours. CONCLUSION: The results illustrate the diagnostic value of CGH in patients with multiple tumours.     

c-fms is present in primary tumours as well as in their metastases in bone marrow.

The expression of c-fms oncoprotein in different primary tumours as well as in their metastases in bone marrow, was shown. All the samples were fixed and processed by the acetone, methyl benzoate, xylene procedure (AMeX), which was suitable for studying oncoprotein expression not only in primary tumours but also in bone marrow (BM) biopsies. Among the patients suffering from acute myeloid leukaemia (AMeL), positive c-fms cells were found in 55% cases. On the contrary, patients with lymphocytic cell disorders have not had detectable c-fms oncogene product in BM biopsies.c-fms oncoprotein was also detected in some primary tumour specimens (lung carcinoma, cervical carcinoma, gastric carcinoma, breast carcinoma and melanoma) and their metastases in BM, while it was not present in normal uterine tissue. There was a positive correlation between c-fms oncoprotein expression in primary and metastatic tumours. Our results showed that c-fms product is confined, not only to some normal, but also to the variety of malignant cells of different origin.     

Morphologie, DNA-Zytophotometrie und Prognose gastrointestinaler neuroendokriner Tumoren (Karzinoide)

A total of 123 manifestations (97 primary tumours and 26 metastases) of neuroendocrine tumours of the gastrointestinal tract observed in 95 patients was investigated for the prognostic value of clinical, histological and DNA cytophotometric parameters. Metastases almost exclusively occurred among ileal carcinoids, which also were responsible for all 14 cases of lethal outcome observed during the follow-up period of mean 42 months. Aneuploid DNA values could be determined significantly more frequently among ileal than in non-ileal carcinoids and showed - upon analysis of the total group of gastrointestinal neuroendocrine tumours - a significant correlation to lethal course of disease. In addition, among 18 cases with primary and secondary carcinoid manifestations available for DNA cytophotometry, an association between the DNA content of metastatic neuroendocrine tumours and prognosis came to light. When applied to the group of ileal neoplasms, however, the parameter DNA content did not allow a better prognostic assessment.     

Secondary neoplasms of the male genital tract with different patterns of involvement in adults and children

AIMS: The incidence, presentation and macroscopic and histological features of secondary neoplasms of the male genital tract are described with reference to their differential diagnosis. METHODS AND RESULTS: A retrospective study of cases from the Royal London Hospital yielded a total of 31 secondary neoplasms involving the testis: 14 at postmortem examination and 17 surgical specimens. Nine cases were leukaemias: six acute lymphoblastic and two acute myeloid leukaemias in children, and one chronic lymphocytic leukaemia in an adult. The commonest primary sites of metastases to the testis were prostate (six cases), stomach (five cases) and lung (three cases). There were two malignant melanomas and isolated examples of metastases from the adrenal gland (neuroblastoma), cerebellum (medulloblastoma), soft tissue (alveolar rhabdomyosarcoma), pancreas and rectum. Of the metastases from solid tumours, 12 involved the right testis only, three involved the left and four were bilateral. In seven of these cases there were multiple testicular nodules, in seven there was a single mass, and in the rest there was diffuse involvement. Secondary neoplasms represented 4.6% of all testicular neoplasms at autopsy, and 1.6% in surgical specimens. There were five secondary penile neoplasms: two each from the pancreas and prostate and one from the bladder. Two neoplasms metastatic to the spermatic cord, both from a gastric primary, were included in the series. CONCLUSIONS: Secondary neoplasms of the testis occur with a frequency comparable to other sites in the genitourinary tract, and metastases to the spermatic cord, epididymis, and penis, are rare in comparison. Disseminated neoplasms rarely present initially at this site and are histologically distinctive in adults, but in children they must be distinguished from primary small round blue cell tumours.     

Growth patterns of pulmonary metastases and primary tumours from five murine fibrosarcoma cell clones.

The growth patterns, including the size, shape and regional preferences, of lung metastases from five murine fibrosarcoma cell clones were studied. Spontaneous metastases developed from tumours formed by subcutaneous inoculation of the cell clones. Lung colonies (experimental metastases) were established by i.v. injection of cells. The numbers of both spontaneously and experimentally formed subpleural lung metastases were counted through a stereomicroscope. The fraction of colonies that was located subpleurally was determined in histological sections of lungs. The growth kinetics of clonally derived primary tumours, and the number of spontaneous and experimental lung metastases, differed greatly between certain cell clones. The number of spontaneous lung metastases was correlated with the maximum size of primary tumours. No close correlation was observed between the size of the primary tumours and the size of experimental metastases. There were differences between the cell clones in the shape and regional preferences of their lung deposits. The subpleural colonies were generally larger than the intrapulmonary ones. Thus, both the regional distribution and the growth pattern of lung deposits differed between the clones.     

An analysis of histology and DNA-ploidy in primary wilms tumors and their metastases and a study of the morphological effects of therapy

Summary In children with Wilms' tumours the length of survival is greatly influenced by success in preventing or controlling metastatic disease. The current study focuses on the morphological aspects of metastases when compared with the primary tumour. In 8 patients it appeared that blastema is the most likely component to metastasize, whereas epithelial and stromal components were hardly, if at all, represented in metastases. Furthermore, flow cytometric DNA ploidy determinations on 4 cases showed that both the primary tumours and the metastases had stemlines in the diploid and low aneuploid (hyperdiploid) range. Finally, in four cases the influence of therapy on morphology of the primary tumours was analyzed. In these cases blastema seemed to be the component most sensitive to therapy. Thus, blastema seems to play a central role in prognosis of Wilms' tumours; either reacting to therapy or, if insensitive, by metastasizing.This study was supported by the Groningen Foundation of Pediatric Oncology and partly by grant GUKC 84-6 of the Netherlands Cancer Foundation (Koningin Wilhelmina Fonds).     

A subset of colorectal carcinomas express c-KIT protein independently of <Emphasis Type="Italic">BRAF</Emphasis> and/or <Emphasis Type="Italic">KRAS</Emphasis> activation

c-KIT is a tyrosine kinase receptor found to be overexpressed in several tumours, namely, GISTs, breast, lung, prostate, ovarian and colorectal carcinomas (CRC). We aimed at determining the frequency of c-KIT expression and mutations in a series of 109 CRC cases (73 primary tumours and 36 lymph node metastases) characterised for KRAS and BRAF mutations. We also aimed at analysing the cellular effects of STI571/Gleevec in CRC-derived cell lines displaying c-KIT expression and KRAS or BRAF mutations. By immunohistochemistry, we found c-KIT overexpression in 15% (11/73) of primary tumours and in 14% (5/36) of metastasis; however, cases showing overexpression did not show c-kit mutations in hotspot regions. The majority (64%) of primary tumours with c-KIT overexpression had mutations at KRAS–BRAF genes. The same was true for 60% of the metastases. We treated CRC cell lines with STI571/Gleevec and verified that it inhibits proliferation and induces apoptosis in all cell lines. In conclusion, overexpression of c-KIT is observed in a subset of primary and CRC metastases in the absence of c-kit mutations. STI571/Gleevec increases apoptosis in CRC cell lines independently of its genetic profile, suggesting that STI571/Gleevec is likely to be an alternative drug for the clinical trials of CRC.     

Predominance of the basal type and HER-2/neu type in brain metastasis from breast cancer.

Although breast cancer is the second most common cause of central nervous system (CNS) metastases with a notable increase of incidence, only few studies on brain-metastasizing breast cancer are available. In this immunohistochemical and fluorescence in situ hybridization (FISH) study, metastases to the CNS (n=85) and primary breast cancers, with known involvement of the CNS (n=44) including paired primary and metastasized tumours (n=23), were investigated retrospectively for the expression of oestrogen- (ER) and progesterone- (PR) hormone receptors, Her-2/neu, epidermal growth factor receptor (EGFR), Ki-67, and cytokeratins (CKs) 5/14. The majority of brain metastases were steroid hormone receptor negative (ER 66%, PR 82%) corresponding to the findings in primary tumours with known involvement of the CNS (68% ER-negative, 75% PR-negative). The frequency of HER-2/neu-overexpressing or -amplified cancers was increased in both groups (34 and 32%, respectively). EGFR expression was more frequent in metastases (41%) than in primary tumours (16%). The proportions of cases with a basal phenotype were 26 and 30%, respectively. In paired primary tumours and metastases to the CNS, constancy of Her-2/neu status was observed in 87% of cases with only one sample turning Her-2/neu-negative and two samples acquiring overexpression/amplification in brain metastases. In contrast, steroid hormone receptors exhibited more frequently a loss of expression (17%) than a gain (9%) with 74% revealing a constant phenotype. We conclude that brain-metastasizing breast cancer belongs predominantly to the basal type or Her-2/neu type. Primary and metastatic tumours differ from each other only in a minority of cases, leading rather to a loss of steroid hormone receptors and to a gain of EGFR and Her-2/neu.     

Significance of the 'maturation' of metastases from germ cell tumours after intensive chemotherapy

A comparison between primary and metastatic germ cell tumours from 38 male patients showed that 19 of 24 metastases with residual differentiated teratoma after adequate therapy came from tumours with teratoma as a component of the primary. The correlation between the presence of teratoma in the primary and the metastases is statistically significant ( P < 0.01) and supports the view that the so called 'maturation' of germ cell tumours is due to selective destruction of anaplastic components in tumours which have already shown an inherent capacity for differentiation. Elevation of the serum concentrations of HCG and AFP on presentation with disseminated disease was significantly related to the presence of morphologically identifiable trophoblast and yolk sac elements respectively in the primary tumours ( P < 0.001). Histological identification and specific mention of teratomatous, trophoblastic and yolk sac elements in reporting germ cell tumours is therefore useful since their presence in the primary correlates with the morphology in the metastases.     

Immunoreactive transforming growth factor alpha and epidermal growth factor in oral squamous cell carcinomas

Forty oral squamous cell carcinomas have been investigated immunohistochemically for the presence of transforming growth factor alpha (TGF-α) and epidermal growth factor (EGF). The same cases were recently characterized for the expression of EGF-receptors. TGF-α was detected with a monoclonal mouse antibody and EGF with polyclonal rabbit antiserum. Thirty-five of the tumours were positive for TGF-α and 26 of the tumours for EGF. None of the poorly differentiated tumours was positive for EGF, but they all were for TGF-α. In sections including normal differentiated oral mucosa, the cells above the basal cell layer were positive for both TGF-α and EGF. The same staining pattern was observed in oral mucosa obtained from healthy persons. In moderately to well differentiated carcinomas, the immunoreactivity was mainly confined to the cytologically more differentiated cells, thus paralleling the situation observed in the normal differentiated oral mucosa. In four cases, material was available from both a primary tumour and a metastasis. Three of these were positive for TGF-α and EGF with the same staining pattern as that of the primary tumours. This investigation together with our previous results confirms the existence of TGF-α, EGF, and EGF-receptors in the majority of oral squamous cell carcinomas and their metastases.     

Metastasis and morphology of transplanted BR6 mouse mammary tumours

BR6 mouse mammary tumours were maintained by serial s.c. transplantation into female or castrated male syngeneic hosts. The tumours could be broadly classified according to their structure, and usually remained stable through many passages. Occasionally, sarcomatous changes were seen (64 out of 800 cases).Pulmonary metastases were found in 5 per cent of mice bearing transplants of well-differentiated acinar tumours, in 34 per cent with tumours of the acinar/ductal type, and in 97 per cent with poorly differentiated tumours. There was a similar correlation between lymph node metastases and degree of primary tumour differentiation, the incidences being 0, 0·5 and 22 per cent respectively. Sarcomatous changes were associated with a reduced incidence of metastasis. Local invasion of muscle or peritoneum occurred in 32 per cent of mice with well-differentiated tumours, and in 59 per cent with less differentiated tumours.Selection for increased metastatic potential was not achieved by transplantation of lung nodules to a subcutaneous site.