The parenchymatous-stromal interrelationships in adenocarcinomas of the large intestine (an electron radioautographic study)

Parenchymatous and stromal cells in colonic adenocarcinomas were studied electron-radiographically. The properties of the nucleic acid and protein synthesis in the cylindrical and goblet cells in carcinoma (partial 3H-thymidine incorporation, heterogeneity of 3H-uridine and amino acid mixture incorporation) indicate a relatively low level of their differentiation, lower degree of their participation in a specific function, retaining their capacity to proliferate. A high number of 3H-thymidine-labelled stem cells which are the main source of tumour growth is observed in carcinoma. Stroma in carcinoma is characterized by endotheliocyte and fibroblast proliferation (3H-thymidine incorporation), high degree of metabolic processes in some of these cells (3H-uridine and amino acid mixture incorporation) and by the disturbance of immunocompetent cell specific function (partial and weak labelling of the immunocompetent cells with 3H-uridine and amino acid mixture).     

Electron microscopic analysis of squamous cell carcinoma of the lung

Twenty-three cases of squamous cell carcinoma of the lungs of various degrees of cataplasia were examined electron microscopically and in 12 cases of these a morphometric analysis of the tumour cell ultrastructure was done. The results showed keratohyalin to be present in squamous carcinoma cells both with and without keratinization; however, its content decreased with the increase of the tumor cataplasia. Thus the degree of development of keratohyalin may be the main objective criterion of the cataplasia degree of squamous cell carcinoma of the lungs. The increased activity of free polyribosomal apparatus with increasing cataplasia of the tumour, increased general synthetic activity of tumour cells as well as the strong correlation of these parameters with the volume fraction of keratohyalin are additional criteria of the degree of cataplasia of squamous cell carcinoma of the lung.     

Comparative evaluation of dysplasia and early cancer of the stomach based on analysis of DNA content

DNA content in the tumour cell nuclei was studied in 16 cases of early stomach carcinoma and in the adjacent areas of the foveolar epithelium dysplasia and metaplastic intestinal epithelium. One wave microspectrophotometry was used; the slides were stained with gallocyanin-chrome alum. The number of DNA synthesizing diploid cells was increasing with the increase of the dysplasia degree. The number of tetraploid cells was higher and the signs of aneuploidy appeared in the areas of noninvasive carcinoma. The stem line in early invasive carcinoma was represented by the diploid cells. The increase of cell atypia in tubular and papillary adenocarcinoma is followed by increase of the tetraploid cell number and by the appearance of polyploidy and aneuploidy. Both undifferentiated and goblet-cell carcinomas have similar parameters of the DNA histogram. DNA content allows an objective judgement on the degree of an early carcinoma cell atypia and differentiation of the advanced dysplasia and noninvasive carcinoma.     

Lysozyme in human gastric carcinoma: a retrospective immunohistochemical study

A total of 171 gastric carcinomas comprising 69 advanced cancers and 102 early cancers were examined immunohistochemically for lysozyme. Tumour cells containing lysozyme were detected in 65 cases or 38% of the 171 gastric cancer cases. The incidence of these cells did not differ remarkably by histological type and infiltrative growth of gastric carcinoma. Of the foregoing 65 cases, two well-differentiated adenocarcinomas and three signet ring cell carcinomas had numerous lysozyme-containing tumour cells, 13 had many argentaffin or argyrophil cells, and 40 had various amounts of several types of mucin. In addition, tumour cells containing both lysozyme and mucin could be identified. No correlation could be observed between lysozyme immunoreactivity in the tumour cells and cellular infiltration of granulocytes or macrophages around the tumour. The lysozyme appeared to be produced by tumour cells. The two year survival rates indicate a tendency for advanced gastric cancers containing lysozyme to have a poor prognosis.     

Demonstration of Insulin production and storage in insulinomas by in situ hybridisation and immunocytochemistry

Twelve cases of insulinoma were studied to assess the amount of hormone synthesis and hormone storage by the tumour and to see what effect a hormone-producing tumour has on the adjacent normal islets. This was investigated by performing in situ hybridization, which detects hormone messenger RNA, thus giving an indication of the degree of hormone synthesis by the tumour, and immunocytochemistry, which detects the hormone itself, thus giving an indication of the amount of hormone stored in the tumour cells. It was found that in most cases there was less hormone stored within the tumour cells than in adjacent islet cells. In a minority of cases, this decrease in stored hormone was associated with reduced hormone synthesis, but the majority of cases showed either equivalent or increased levels of hormone mRNA in the tumour cells compared with adjacent islets. In addition, it was noted that, unlike some other endocrine organs, the presence of a hormone-producing tumour within the pancreas did not appear to inhibit hormone synthesis in the adjacent normal tissue. © 1997 John Wiley & Sons, Ltd.     

Effects of prolonged treatment with decarbazine on tumour metastatic potential in mice bearing Lewis lung carcinoma

The effects of decarbazine on tumour growth and metastatic dissemination upon treatment protracted for 10 tumour transplant generations were examined in mice bearing Lewis lung carcinoma. Primary tumour growth is unaffected by the drug, independently from the duration of the treatment. In contrast, dacarbazine significantly inhibits the formation of lung metastasis. The proportion of mice with metastasis decreases for an increasing number of transplant generations of treatment, and after 10 transplant generations of treatment metastatic capacity is completely lost in immunocompetent mice. The reduction in metastatic potential is relatively stable, being retained for three successive transplant generations without treatment. The metastatic potential of treated tumours in immunosuppressed mice is substantially similar to that in immunocompetent hosts, indicating that chemical xenogenization of tumour cells does not occur as reported for transplantable mouse leukaemias. The results obtained using clonally selected tumour lines with different metastatic potential rule out the selection by dacarbazine of tumour cell sublines with reduced metastatic potential as the mechanism of the drug's action. Upon prolonged treatment, dacarbazine appears to cause a rather stable and dramatic loss in metastatic potential, not accompanied by resistance, which might be attributed to genotypic alteration(s) of tumour cells, and which might participate into the clinical effects of the drug.     

Lymphoepithelioma-like carcinoma of the breast: lack of evidence of Epstein-Barr virus infection

AIMS: Six cases of lymphoepithelioma-like carcinomas of the breast were studied in order to better define the morphology of this type of breast tumour and to see if these neoplastic lesions are related to Epstein-Barr virus (EBV) infection by analogy with histologically similar tumours located in other organs. METHODS and RESULTS: The cases were studied with immunohistochemistry using several antibodies and with in-situ hybridization method employing different types of nucleotides in four cases. The cases presented the distinctive morphology of the lymphoepithelioma-like carcinomas as seen in other organs. Specifically Rigaud and Schminke's patterns of growth were observed. In addition, at the edge of the neoplastic lesions, a lymphoepithelial lobulitis was seen. In-situ hybridization failed to reveal any positivity for EBV genome. CONCLUSION: Lymphoepithelioma-like carcinoma is a specific type of breast tumour, but its relation to EBV has yet to be proven. The differential diagnosis with medullary carcinoma is discussed.     

Organ-Specific Membrane Antigens: Attempts to Produce Specific Antisera for Mixed Antiglobulin Tests on Disaggregated Cells

A satisfactory system for testing the reaction of rabbit antisera with membrane antigens of human tissue cells is described. This method allows the differentiation between IgG and IgM antibodies and provides an extremely sensitive method for the detection of antigens on all cells including non-viable fixed cells.Anti-organ serum before selective absorption showed very little organ specificity in their reactions, but may be made specific by extensive absorption although often the resulting specific titre was very low.Organ-specific membrane antigens were also identified and shown to be represented on tumour cells, although in some cases such as the colon the reactions were weaker with tumour cells than with normal parenchymal cells of an organ. On the other hand, in one case of carcinoma of the kidney the organ-specific antigens were detectably stronger on tumour cells than on normal kidney cells.Preliminary studies on human ascitic tumour cells from 4 different cancer patients show that species-specific membrane antigens can be demonstrated. Unfortunately none of the cases were derived from organs whose origin could be identified with the antisera which had been prepared for this series of experiments.     

Comparative electron microscopic study of cancer cell differentiation in the primary tumor and the metastases

Comparative ultrastructural analysis of primary carcinomas of the lung, mammary gland, colon, stomach and some other sites and their metastases into the lymph nodes and inner organs is presented. It is established that the main ultrastructural specific features of differentiation typical for the cells of primary tumours are, as a rule, retained in their metastases to various organs and tissues. Carcinoma cells in the metastases repeat most frequently (81% of cases) the types of differentiation of a primary tumour. However, the reduction of a number of differentiated cell types (5%) or appearance of a new ultrastructural cell type (14%) may be found in the secondary tumour deposits. Besides, the retention (74%), decrease (10%) or increase (16%) of the degree of differentiation may be observed in metastases as compared to the primary tumour. The data obtained may be the ground for a proper identification of malignant tumours on the basis of the electron microscopic study of their metastasis.     

Tumour regression and mesorectal lymph node changes after intensified neoadjuvant chemoradiation for carcinoma of the rectum

Neoadjuvant radiation or chemoradiation is currently the treatment of choice for patients with locally advanced carcinoma of the rectum. To assess the effects of chemoradiation on tumour regression and on uninvolved mesorectal lymph nodes, a consecutive series of 76 patients receiving neoadjuvant chemoradiation and a stage-adapted control series of 57 patients without pretreatment were studied. Densities of cells positive for CD4 (T-helper cells), CD8 (cytotoxic T-cells), CD83 (mature dendritic cells), and CD57 (natural killer cells) were determined on immunostains. Tumour regression was graded, and presence or absence of extramural tumour was recorded. The densities of CD4+ T-lymphocytes and CD83+ dendritic cells in the paracortex of mesorectal lymph nodes were observed to be significantly reduced, as were the densities of CD57+ cells in the follicles; densities of CD8+ T-lymphocytes did not differ. Strong, moderate and poor tumour regression was observed in 29, 25, and 22 cases, respectively. For 12 patients, absence of extramural vital or regressing tumour was recorded, indicating pretherapeutic overstaging. The results bring to mind that neoadjuvant chemoradiation as a side effect may have a negative impact on anti-tumour immunity. Together with the drawback of overstaging the results argue for a careful selection of patients.     

Enhanced thermal stability of lysosomal beta-D-galactosidase in parenchymal cells of tumour bearing mice.

The thermal stability of the enzyme beta-D-galactosidase varies among different organs in normal C57Bl/6 mice, and increases in the same organs in mice with Lewis Lung carcinoma. Thermal stability of this enzyme is also increased by treatment of the mice with cell-free extracts of tumour cells or with inflammatory compounds such as carrageenan or orosomucoid. After desialylation, orosomucoid more effectively increases the heat stability of the enzyme. By contrast talc, which has no galactosyl groups, is without effect on the stability of the enzyme in vivo. Macrophages of tumour bearing mice release into the culture medium a more heat resistant enzyme than macrophages from control mice. In both cases the heat resistance of the secreted enzyme is higher when fetal calf serum is present in the culture medium. Bovine serum does not modify the thermal stability of beta-D-galactosidase in this system. Incubation of lysosomal fractions of various organs with the synthetic beta-D-galactosidase substrate, p-nitrophenyl-galactopyranoside, also strongly increases the heat resistance of the enzyme. The results suggest that one factor influencing the heat resistance of this enzyme may be complex formation between the enzyme and its substrates, an example of substrate protection of the enzyme. This may not be the only factor involved in enzyme stabilization in vivo.     

Flow cytometry of DNA in research on kidney tumors

The DNA content was studied in 22 patients (including 10 children) with renal tumours. Renal clear cell carcinomas were mainly diploid (5 of 6 cases); one clear cell and other variants of renal cell carcinoma were aneuploid. There was a correlation between the degree of tumour cell anaplasia and ploidy of renal cell carcinomas: all diploid carcinomas were of I and II degree of anaplasia, aneuploid carcinomas, except one case, were of degree III of anaplasia. 5 out of 9 nephroblastomas were diploid, 4, aneuploid. The distinctive features of nephroblastoma were pronounced proliferative activity of tumour cells and a low DNA index of the aneuploid cell line which in all cases was localized in the vicinity of the diploid region. The remaining tumours (papillary epithelial nephroma, juxtaglomerular cell tumour and malignant schwannoma) were diploid with a relatively low proliferative activity of tumour cells.     

Enhanced thermal stability of lysosomal beta-D-galactosidase in parenchymal cells of tumour bearing mice

The thermal stability of the enzyme beta-D-galactosidase varies among different organs in normal C57Bl/6 mice, and increases in the same organs in mice with Lewis Lung carcinoma. Thermal stability of this enzyme is also increased by treatment of the mice with cell-free extracts of tumour cells or with inflammatory compounds such as carrageenan or orosomucoid. After desialylation, orosomucoid more effectively increases the heat stability of the enzyme. By contrast talc, which has no galactosyl groups, is without effect on the stability of the enzyme in vivo. Macrophages of tumour bearing mice release into the culture medium a more heat resistant enzyme than macrophages from control mice. In both cases the heat resistance of the secreted enzyme is higher when fetal calf serum is present in the culture medium. Bovine serum does not modify the thermal stability of beta-D-galactosidase in this system. Incubation of lysosomal fractions of various organs with the synthetic beta-D-galactosidase substrate, p-nitrophenyl-galactopyranoside, also strongly increases the heat resistance of the enzyme. The results suggest that one factor influencing the heat resistance of this enzyme may be complex formation between the enzyme and its substrates, an example of substrate protection of the enzyme. This may not be the only factor involved in enzyme stabilization in vivo.     

Morphological evidence for field effect as a mechanism for tumour spread in mammary Paget's disease

AIMS: The histogenesis of mammary Paget's disease is controversial. The purpose of this study was to investigate the mechanism of tumour spread in the nipple epidermis by examining 28 cases of mammary Paget's disease associated with underlying intraductal carcinoma. METHODS AND RESULTS: The atypical cells in the epidermis displayed a spectrum of cytological changes ranging from small-sized atypical cells located in the basal cell layer to large-sized atypical cells characteristic of Paget's cells in the upper layer of the epidermis. Serial sectioning revealed the presence of isolated, scattered and small atypical cells in the basal cell layer at the periphery of the epidermal lesion. The atypical cells, including those in the basal cell layer showed positive immunostaining for cytokeratin 7 and Her2/neu oncoprotein. Electron microscopy examination demonstrated the presence of intercellular junctions of desmosomal-like or desmosomal types between tumour cells and adjacent squamous cells. Furthermore, examination of the intraductal carcinoma of the breast tissue in cases of Paget's disease as well as control cases of intraductal carcinoma also revealed areas of skip lesions of intraductal carcinoma. CONCLUSIONS: In view of these changes, it is unlikely that tumour expansion or tumour cell motility are sufficient explanations to account for the pattern of tumour spread in both the epidermis and the duct epithelium with skip lesions. A 'field effect' in the duct system harbouring intraductal carcinoma and the adjacent epidermis may play an important role in the tumour cell spread in the epidermis as well as in the ductal epithelium.     

Lymphatic vascular density and lymphangiogenesis during tumour progression of carcinoma ex pleomorphic adenoma

AIMS: To assess lymphatic vascular density (LVD) and lymph vessel endothelial proliferation in a series of carcinoma ex pleomorphic adenoma (CXPA) that represents the tumour in the different carcinogenesis phases and tumour progression. METHODS: In 8 cases of early CXPA (intracapsular and minimally invasive tumours), 8 of advanced CXPA (widely invasive tumours) and 10 of pleomorphic adenoma (PA) without malignant transformation, lymphatic vessels and proliferating cells were detected using the antibodies D2-40 and Ki-67 respectively. RESULTS: Comparing early tumours with advanced ones, LVD was not significantly different at the tumour margin. In contrast, regarding intratumoural lymphatics, PA without malignant transformation and early CXPA contained rare, if any, lymph vessels, whereas in widely invasive carcinomas they were more numerous. However, neither intratumoural nor peritumoural LVD were increased in comparison to adjacent normal salivary gland tissue. In no case did dual immunohistochemistry using D2-40 and the cell proliferation marker Ki-67 reveal the existence of proliferating lymphatics. Carcinomatous emboli were found in peritumoural as well as in intratumoural lymphatics only in advanced CXPA without myoepithelial differentiation. CONCLUSION: In CXPA, the lymphatic network is mainly composed of pre-existing lymphatics which are rare in tumours that have not infiltrated outside the confines of the original PA. In the widely invasive CXPA, intratumoural as well as peritumoural lymphatics are a conduit for carcinoma cells, but in carcinomas with myoepithelial differentiation, the neoplastic cells seem to have a lower invasion capacity.     

Expression of amphiregulin and epidermal growth factor receptor in human breast cancer: analysis of autocriny and stromal-epithelial interactions

Amphiregulin (AR) and its receptor, epidermal growth factor receptor (EGFR), were evaluated by dual immunostaining in a series of 84 invasive ductal breast carcinoma specimens, 33 of which were from locally advanced inflammatory (T4d) cancer. Co-expression of AR and EGFR was always found in non-malignant breast tissues adjacent to tumours (24/24). Alternatively, expression of AR and EGFR was found in invasive epithelial tumour cells in 50% and 17.8% of specimens, respectively. In tumour stroma, 59.5% and 30.9% of specimens, respectively, were positively stained. By univariate analysis, AR and EGFR expression in invasive carcinomas was correlated with large tumour size, inflammatory carcinoma, node involvement, Bloom-Richardson (SBR) grade III, and absence of oestrogen receptor. EGFR expression in stromal cells was correlated with non-inflammatory carcinoma. A putative autocrine loop with AR and EGFR expression in invasive carcinoma was detected in 14.3% of cases. Stromal expression of AR and EGFR expression in invasive tumour cells was detected in 11.9% of cases and related to poor prognostic parameters. By multivariate analysis, AR expression in invasive tumour was strongly related to inflammatory carcinoma (p=0.005) and marginally related to SBR grade III (p=0.07). EGFR expression in invasive tumour and stromal cells was correlated with absence of oestrogen receptor and non-inflammatory carcinoma (p=0.002 and p=0.015, respectively).     

Extracellular matrix in preneoplastic lesions and early cancer of the lung

Characterization of preneoplastic lesions mainly concentrated on cellular, nuclear and epithelial atypias. The extracellular matrix was almost neglected, although malignant tumour cells interact with extracellular matrix molecules during tumour invasion. Until now systematic investigations of extracellular matrix components in preneoplastic lesions and early stages of lung cancer are lacking. 150 preneoplastic lesions, 10 specimens of early cancer and 30 specimens of normal bronchial mucosa were examined by means of immunofluorescence microscopy. The distribution of collagen type I and III and the non-collagenous glycoproteins laminin and fibronectin have been investigated by an indirect immunohistochemical method. With an increasing degree of preneoplasia an increased matrix disarrangement of the basement membrane zone could be observed. In severe dysplasia and carcinoma in situ neosynthesis of collagen type III and especially laminin in close association to neoangiogenesis could be demonstrated besides a disintegration of extracellular matrix components. In early lung cancer numerous laminin positive basement membrane like structures are situated around tumour cells. An enhanced deposition of collagen type I and III fibres could be demonstrated around tumour cell islets. The results indicate a partial loss of function in preneoplastic basal cells in cases of dysplasia and carcinoma in situ. In early stages of invasive squamous cell carcinoma of the bronchus, extracellular matrix components obviously could be produced by tumour cells.     

A major solid undifferentiated carcinoma pattern correlates with tumour progression in locally advanced prostatic carcinoma

Solid undifferentiated carcinoma was the major microscopic pattern in 24 prostatic carcinomas, 12 of which were clinically recurrent. Tumour cells were uniform, with moderately hyperchromatic nuclei and indistinct cytoplasm, and were arranged in solid or focally irregular aggregates. In areas, the tumour cells were large with vesicular nuclei, nucleoli and more abundant cytoplasm. In previous specimens, solid undifferentiated carcinoma was absent or was a minor pattern. Twenty of 23 cases showed prostate specific antigen and prostatic acid phosphatase immunoreactivity, and nine of 17 cases contained scattered argyrophilic or chromogranin-immunoreactive cells. On proliferating cell nuclear antigen immunostaining of 12 specimens, the mean tumour proliferative fraction in solid undifferentiated carcinoma (range: 10.5-18%) was greater than in areas of grade 3 prostatic carcinoma (range: 3-6%). In all 22 stage C and D cases, there was a close correlation with clinical evidence of tumour progression, and the overall 2-year survival rate was only 16.7%. It is concluded that a major solid undifferentiated pattern correlates with increased biological aggressiveness and a poor prognosis in locally advanced prostatic carcinoma.     

Expression of EDA/EDB isoforms of fibronectin in papillary carcinoma of the thyroid

Cellular fibronectins containing the extracellular domain A or B (EDA and EDB) are particularly abundant in fetal and neoplastic tissues. The presence of EDA and EDB was investigated in 28 cases of papillary carcinoma of the thyroid using IST-9 and BC-1 monoclonal antibodies. Immunostaining for EDA and EDB was detected in tumour stroma, in tumour basement membranes, and in tumour blood vessels. EDA was present in 27 of the 28 cases, in 20 of which more than 75 per cent of the tumour stroma was stained. Immunostaining for EDB was detected in 23 of the 28 cases and was less pronounced than that for EDA, being present in less than 25 per cent of the tumour stroma in most cases. Reactivity for EDA/EDB was not observed in the adjacent normal thyroid in any of the cases investigated. In a group of 20 non-papillary tumours, immunostaining for EDA was present in the stroma of three follicular carcinomas (one minimally and two widely invasive), one medullary carcinoma, and 5 of 16 follicular adenomas; expression of EDB was more restricted, being present in only the two cases of widely invasive follicular carcinoma. The presence of EDA and EDB was not correlated with the extent of fibrosis or the degree of tumour cell differentiation. Immunoreactivity was already present in microcarcinomas. These observations raise the possibility that the production of oncofetal fibronectins is an important step in papillary carcinoma tumourigenesis, perhaps facilitating adhesion and spreading of tumour cells. Copyright © 1999 John Wiley & Sons, Ltd.     

Basaloid-squamous carcinoma of the upper aerodigestive tract and so-called adenoid cystic carcinoma of the oesophagus: the same tumour type?

Basaloid-squamous carcinoma of the larynx, pharynx and base of tongue and the so-called adenoid cystic carcinoma of the oesophagus are rare but distinctive tumours associated with a grave prognosis. They occur most commonly in elderly males and present at an advanced stage. Our study of four such laryngeal tumours and five such oesophageal tumours shows that they are histologically and immunohistochemically identical, providing support for the idea that they are the same tumour type. They show a biphasic pattern in which basaloid tumour is intimately associated with a neoplastic squamous component which can be invasive or in situ. The basaloid component is in the form of invasive lobules with frequent comedo-necrosis and hyalinization. The constituent cells possess pale pleomorphic nuclei with frequent mitoses. Immunoreactivity for cytokeratin in the basaloid component is remarkable for its absence or weak and focal nature. Review of the literature shows that only a few cases of 'adenoid cystic carcinoma' of the oesophagus are bona fide examples of adenoid cystic carcinoma as it occurs in the salivary glands, while the others are identical to basaloid-squamous carcinoma of the upper aerodigestive tract. Their distinction is important because genuine adenoid cystic carcinoma is much less aggressive than basaloid-squamous carcinoma.