Amyloid deposits in basal cell carcinoma of the skin. A pathologic study of 199 cases.

Deposits of amyloid were detected in 101 of 199 basal cell carcinomas (51%). The frequency of amyloid deposits in solid, adenoid, and cystic histologic subtypes was slightly higher than overall, whereas in partial sclerosing and morphea-like tumors the frequency was much lower. The amyloid of basal cell carcinoma showed histochemical characteristics that were different from those of locally deposited amyloid in endocrine tumors such as medullary carcinoma thyroid and from those of "secondary" amyloid. No major differences in the histochemical characteristics, however, were observed between amyloid associated with basal cell carcinoma and myeloma-associated or "primary" amyloid. Nevertheless, immunohistochemical staining with rabbit antihuman keratin antibodies by the peroxidase antiperoxidase technique demonstrated positivity only in amyloid deposits associated with basal cell carcinoma and not in those of myeloma-associated amyloid. This evidence supports the concept that amyloid of basal cell carcinoma is produced in the stroma from degenerated epithelial cells through filamentous degeneration or apoptosis.

Constitutive absence and interferon-gamma-induced expression of adhesion molecules in basal cell carcinoma.

Adhesion of lymphocytes to target cells via certain cell surface molecules is important in cytotoxic T lymphocyte-mediated immune reactions. The binding of lymphocyte function-associated (LFA) antigens 1 and 2, with their respective ligands, intercellular adhesion molecule-1 (ICAM-1) and LFA-3, which are expressed on the surface of nonlymphoid cells, has been shown to be critical for lymphocyte adhesion. To determine whether basal cell carcinomas (BCCs) can escape immunodetection as a result of the inability of cytotoxic T lymphocytes to bind tumor cells, the expression of adhesion molecules on numerous BCCs, before and after exposure to interferon-gamma (IFN-gamma), was examined. Ninety-three percent of 30 freshly excised invasive BCCs did not express ICAM-1 and 73% of 11 BCCs did not express LFA-3. However, the normal-appearing basal keratinocytes in epidermis overlying nests of BCC, did express ICAM-1, particularly when a marked LFA-1+ and LFA-2+ dermal lymphocytic infiltrate was present. After BCC tissue was incubated in vitro with IFN-gamma the expression of ICAM-1 was induced on 85% of tumors studied. Thus tumor cells did not possess an absolute inability to express adhesion molecules; rather the constitutive absence of such molecules may be due to insufficient in vivo cytokine levels necessary to induce expression or a barrier preventing cytokines from reaching and interacting with tumor cells. We conclude that the absence of ICAM-1 and LFA-3 adhesion molecules is a mechanism by which BCCs can avoid immunosurveillance.

Localization of integrin receptors for fibronectin, collagen, and laminin in human skin. Variable expression in basal and squamous cell carcinomas.

VLA integrins in human skin were examined by indirect immunofluorescence utilizing antibodies recognizing the beta 1, alpha 2, alpha 3, or alpha 5 subunits. Staining of fetal, newborn, or adult skin with antibodies to beta 1, alpha 2, or alpha 3 subunits gave essentially similar staining patterns: intense staining was associated with the basal layer of the epidermis, hair follicles, and blood vessel walls. The alpha 5 subunit could be detected only in epidermis and the inner root sheath of hair follicles in fetal skin. In epidermis, the staining reaction for the beta 1 subunit was not only found in sites interfacing with the basement membrane zone, but also around the entire periphery of these cells. We speculate that these receptors might have previously unrecognized functions in cell-cell interactions or that these findings may suggest the presence of previously unrecognized ligands in the intercellular spaces of keratinocytes. Examination of nine nodular basal cell carcinomas revealed a prominent staining reaction with anti-beta 1 and anti-alpha 3 antibodies at the periphery of the tumor islands. In contrast, staining of five squamous cell carcinomas revealed either the absence of integrins or altered and variable expression. Thus, matrix components and their receptors may participate in modulation of growth, development, and organization of human skin.

Distribution of neu (c-erbB-2) protein in human skin.

The neu (c-erbB-2) gene encodes a transmembrane protein with tyrosine kinase activity that appears to be a growth factor receptor. Antibody was generated by immunization of rabbits with a synthetic polypeptide that was based on an internal sequence at the carboxy terminus of the molecule. This antibody was used to survey the expression of neu in human skin by immunohistochemistry. Significant protein was found in the squamous cell layer of the surface epidermis, in squamous cell carcinomas, in the external root sheath of hair follicles, and in eccrine gland secretory cells; it was poorly expressed in the basal cell layer and in a basal cell carcinomas. Increased neu expression appears to be associated with the differentiation of keratinocytes.

Identification of the origin of cells in human basal cell carcinoma xenografts in mice using in situ hybridization.

In an attempt to identify more clearly the origin of cells in human basal cell carcinoma xenografts in mice, paraffin and frozen sections were subjected to in situ DNA hybridization with biotin labelled human and murine DNA probes. Human skin and mouse skin sections were used as controls. As expected, the implanted epithelium reacted with the human DNA probe and the surface epithelium and most of the stromal cells reacted with the murine probe. However, the stroma immediately surrounding the implanted epithelium contained cells of human origin mixed with murine cells. Occasional murine cells (presumed inflammatory) were present in the human implanted epithelium. Assessment showed no correlation between the degree of differentiation of the implanted epithelium and the ratio of human/murine cells in the contiguous stroma. This technique provides a sensitive test for identifying human cells in xenografts and may be useful in assessing the role of stromal cells in the differentiation of a variety of carcinomas.

An absence of human leukocyte antigen-DR and a decreased expression of beta 2-microglobulin on tumor cells of basal cell carcinoma: no influence on the peritumoral immune infiltrate.

The expression of human leukocyte antigen-DR (HLA-DR) and beta 2-microglobulin on the tumor cells and their correlation (if any) to the degree and the composition of the peritumoral mononuclear infiltrate were studied in 37 basal cell carcinomas from 32 patients with an indirect immunoperoxidase technique. In 36 of 37 basal cell carcinomas (97%) there was no expression of HLA-DR on tumor cells of basal cell carcinoma. In 13 of 37 basal cell carcinomas (35%) beta 2-microglobulin was expressed on the tumor cells. Both a diffuse cytoplasmic and a membrane staining were observed in only six of these 13 basal a diffuse cytoplasmic and a membrane staining were observed in only six of these 13 basal cell carcinomas; in the other seven basal cell carcinomas only a diffuse cytoplasmic staining was observed. In all 37 basal cell carcinomas there was membrane staining for beta 2-microglobulin in the normal epidermis. The intensity of staining in the normal epidermis was always stronger than that in the tumor nests. There was a varying degree of peritumoral immune infiltrate in all basal cell carcinomas. It comprised mainly T cells (mean percentage 57 +/- 15). In the group of patients with basal cell carcinoma with moderate to heavy infiltrate the mean percentage of T cells was 63 +/- 13, which was significantly higher than the mean percentage of T cells (46% +/- 14%) in the group of patients with basal cell carcinoma with a mild infiltrate. This difference was mainly the result of an increase in T helper cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Human papilloma virus infection and skin cancer in renal allograft recipients.

202 renal allograft recipients in south-east Scotland, who had received transplants between 1965 and 1986, were monitored over 3 years (1984-87) for the presence of warts, keratoses, and skin cancers. 77% of 69 patients with graft survival of more than 5 years had viral warts, 38% had keratoses, and 12% had skin cancers, whereas of the 133 with graft survival of less than 5 years 20% had warts, 17% had keratoses, and 1.5% had skin cancers. The ratio of squamous cell carcinoma to basal cell carcinoma was 15:1. Most viral warts showed significant epidermal dysplasia, and keratoses and squamous cell carcinomas had signs of human papilloma virus infection. 15 (60%) of 25 squamous cell carcinomas contained HPV5/8 DNA and 1 contained HPV4 DNA--HPV5/8 DNA was detected in skin lesions of recipients with cancers significantly more often than in those matched for duration and type of immunosuppression with nonmalignant skin lesions. The findings suggest a role for HPV5/8 in the aetiology of squamous cell carcinoma in renal allograft recipients.

Immunoelectron microscopic studies on cytokeratins in human basal cell carcinoma.

Ultrastructural investigations into the location and the expression of the cytokeratins 7, 8, 10 and 19 were undertaken on ultrathin cryosections of 8 basal cell carcinomas (BCC) using a high resolution immunogold labeling method and five different monoclonal antibodies against specific cytokeratins. The results showed that cytokeratin 10 was expressed only in the differentiated keratinocytes. In contrast to the previously reported biochemical and immunohistochemical studies, in this study cytokeratin 19 was expressed not only in the tumor cells of BCC but also in the normal epidermal keratinocytes. The expression of cytokeratin 7 in BCC could not be confirmed but the lack of expression of cytokeratin 8 was confirmed, excluding its potential role as a specific histopathological marker for BCC.

[Immunohistochemical study of the distribution of carcinoembryonic antigen and keratin in tumors of epidermal origin. I: Squamous and basal cell epitheliomas]. / Estudio inmunohistoquímico de distribución de antígeno carcinoembrionario y queratina en tumores de origen epidérmico. Parte I: Epiteliomas espinocelulares y basocelulares.

Carcinoembryonic antigen (CEA) has been studied extensively, in association with visceral tumors and normal tissues. Although CEA has been demonstrated in sweat gland cutaneous tumors, little is known about its presence in skin and epithelial related tumors. We studied the presence of CEA and keratin in squamous cell and basal cell carcinoma using immunohistochemical techniques. Its distribution related with differentiation, proliferation and malignant potential is observed. The expression of both antigens is correlated.

Distribution of desmoplakin in normal cultured human keratinocytes and in basal cell carcinoma cells.

In cultured human keratinocytes (NHEK) maintained in medium containing low levels of Ca2+ (0.04 mM) desmoplakin is a component of certain electron-dense bodies in the cytoplasm. These bodies are associated with bundles of intermediate filaments. Upon elevation of the level of Ca2+ in the culture medium to 1.2 mM, desmoplakin first appears at sites of cell-cell contact in association with bundles of intermediate filaments. Subsequently, desmoplakin becomes incorporated into desmosomes in a manner comparable to that seen in mouse keratinocytes (Jones and Goldman: Journal of Cell Biology 101:506-517, 1985). NHEK cells maintained for 24 hr at Ca2+ concentrations between 0.04 mM and 0.18 mM were processed for immunofluorescence, immunoelectron, and conventional electron microscopical analysis. In NHEK cells grown at Ca2+ concentrations of 0.11 mM, desmoplakin appears to be localized in electron-dense bodies associated with intermediate filaments at sites of cell-cell contact in the absence of formed desmosomes. At a Ca2+ concentration of 0.13 mM desmoplakin is arrayed like beads on a "string" of intermediate filaments at areas of cell-cell association. At 0.15 mM, desmosome formation occurs, and desmoplakin is associated with the desmosomal plaque. In basal cell carcinoma cells desmoplakin is not restricted to desmosomes but also occurs in certain electron-dense bodies morphologically similar to those seen in NHEK maintained in low levels of Ca2+ and during early stages of desmosome assembly. We discuss the possibility of "cycling" of desmoplakin through these bodies in proliferative cells.

[Prostacyclin (PGI2) biosynthetic activity in variously differentiated neoplasms and the surrounding tissue]. / Zur Prostazyklin- (PGI2-) Biosyntheseaktivität in unterschiedlich differenzierten Neoplasien und umgebendem Gewebe.

For further elucidation of the connection between tumor differentiation, intensity of stroma reaction and the capacity of prostacyclin biosynthesis comparable investigations were performed on the PGI2 formation and the microscopic pattern of 10 tissue samples from naevus cell tumors, skin basalioma, oral pavement epithelium carcinoma and their direct adjoining region. The effects were compared with 10 tissues from healthy oral mucosa and facial skin. The results indicate that the PGI2 activity depends on the quantity of cells within the tissue. The various cells were stimulated in a different manner. The highest values of the PGI2 biosynthesis were found in cells of an inflammatory proliferation direct adjoining the oral pavement epithelium carcinoma. This shows a connection between tumor differentiation, intensity of stroma reaction and the activity of prostacyclin biosynthesis, possibly indicating a criterion of malignity for the prognosis of the tumor disease.

Fibronectin gene expression by epithelial tumor cells in basal cell carcinoma: an immunocytochemical and in situ hybridization study.

Previous observations have demonstrated that fibronectin is deposited in high abundance in basal cell carcinoma stroma. In this study, the nature of fibronectin and the site of its synthesis were explored in 10 basal cell carcinomas of the nodulo-ulcerative type by immunocytochemistry and in situ hybridization. First, simultaneous localization of epithelial tumor cell islands and fibronectin epitopes was carried out by double immunofluorescence staining with monoclonal anti-cytokeratin antibodies and polyclonal fibronectin antibodies, the latter recognizing both the cellular and plasma types of the protein. Large amounts of fibronectin were deposited in the basal cell carcinoma stroma, with the highest concentration present in the immediate proximity of the epithelial cell islands. Immunofluorescence with a monoclonal anti-fibronectin antibody, which is directed against the ED-domain of cellular fibronectin and does not recognize the plasma type of fibronectin, revealed essentially the same staining pattern as that obtained with the polyclonal anti-fibronectin antibody. This observation suggested that fibronectin in BCC was predominantly of the cellular type. Second, in situ hybridizations, utilizing a human fibronectin specific cDNA, demonstrated that the highest concentration of fibronectin mRNA was found in the most peripheral cell layer of the epithelial tumor islands. The presence of fibronectin mRNAs in the tumor cells of the central regions of the islands, as well as within occasional stromal cells, was also noted. Thus, two lines of evidence suggest that the epithelial tumor cells are predominantly responsible for the synthesis and deposition of fibronectin in basal cell carcinoma. The presence of fibronectin may explain the characteristic biologic behavior of basal cell carcinomas, including low degree of metastatic potential and local destructive nature of the tumors.

Determination of sex steroid receptor in human basal cell carcinoma.

The role of estrogens in the development of skin cancer is controversial. Sex steroids have a profound effect on the epidermis and epidermal appendages. Estradiol in pharmacologic doses has been reported to stimulate basal cell carcinoma in an animal model. Sex hormones act by means of a specific protein receptor. In this study we used a specific, highly sensitive monoclonal antibody to evaluate sex steroid receptors in human basal cell carcinoma. No estrogen or progesterone receptor protein was detected in the basal cell tumor, despite clear positive control tissues. We conclude that these sex steroid receptors are not present in significant amounts to mediate a direct effect in basal carcinoma.

Distribution of actin filaments in human malignant keratinocytes.

Distribution of actin filaments in human malignant keratinocytes was examined by immunofluorescence staining. The primary cultures were obtained from a squamous cell carcinoma, a basal cell carcinoma, and Bowen's disease. Rhodamine-phalloidin staining revealed that actin filaments were occasionally organized to form stress fibers, many short bundles with a ripple appearance, and regular arrays of actin patches. Some of these structures appeared in untransformed keratinocytes as a result of a brief exposure to a tumor promotor, TPA. These findings suggest that regulation of actin functions is involved in neoplastic processes from the very early stages and that alteration is persistent in neoplastic cells.

Detection of transforming growth factor alpha in normal, malignant, and hyperproliferative human keratinocytes.

Transforming growth factor alpha (TGF-alpha) is a 50-amino acid peptide, previously demonstrated only in transformed cell lines and human tumors, which is structurally homologous to epidermal growth factor (EGF). TGF-alpha expression in keratinocytes from normal individuals, patients with psoriasis, and patients with malignant skin diseases was investigated using an mAb raised against synthetic human TGF-alpha. mAb A1.5 reacted with TGF-alpha, but not EGF, in a sensitive ELISA. Keratinocytes in eight nodular basal cell carcinomas, one morpheic basal cell carcinoma, and one squamous cell carcinoma demonstrated intense membranous immunoperoxidase staining with mAb A1.5. Of even greater interest was the observation that the overlying normal epidermis, as well as the epidermis from five normal skin specimens, were stained by the mAb. Keratinocytes in plaques from 18 psoriasis patients were more intensely stained than those from normal skin. Cultured normal keratinocytes demonstrated membranous staining with mAb A1.5. Absorption of mAb A1.5 with synthetic human TGF-alpha completely removed the reactivity of mAb A1.5 with both basal cell tumors and normal epidermis. The demonstration of TGF-alpha in normal keratinocytes suggests that it plays a role in normal keratinocyte growth, wound healing, and in the pathogenesis of acanthosis.

[Collagen type IV of the basal membrane of the epithelium of the skin in certain forms of pathology]. / Kollagen IV tipa bazal'noi membrany épiteliia kozhi pri nekotorykh formakh patologii.

Using immunofluorescence, attenuation or complete disappearance of type IV collagen antiserum reaction were shown in the epidermal basal membrane zone of SLE patients. Some positive material was revealed in the intercellular spaces of the epidermis. There is an inverse correlation between the amount of immune complexes in the dermo-epidermal region and the intensity of anticollagen serum reaction in the basal membrane zone. This can be explained by the toxic effect of immune complexes on collagen-synthesizing cells and the disturbance of this protein fixation in the basal membrane followed by its sequestration from the affected regions. Complex therapy including hemoperfusion restores normal collagen location in the epidermal basal membrane.

Expression of keratin proteins in deeply invasive basal and squamous cell carcinoma: an immunohistochemical study.

The expression of certain classes of keratin is associated with cell maturation and differentiation. During cell transformation and tumor development, the cell specificity of intermediate filament, keratin, is largely conserved. Taking advantage of this, we utilized monoclonal antikeratin immunohistochemical techniques to examine basal and squamous cell carcinomas as they became deeply invasive. Dyskeratotic keratinocytes and keratin pearls in squamous cell carcinomas (SCCs) stain with antikeratin antisera to larger keratins (65-67 Kd). At the deep tumor margins, SCCs no longer express larger keratins but retain expression of 50, 58 Kd, which are markers of keratinocytes derived from stratified squamous epithelial cells. This selective loss of expression of keratin polypeptide markers of differentiation in SCC is associated with progressively more aggressive biologic behavior as the tumor invades deeper structures such as muscle and bone. Recurrent basal cell carcinoma (BCC) which was of the nodular type when first excised, shows features of morphea-like BCC and of aggressive growth patterns at the deep invasive margin. At these deep margins, some tumors express markers of keratinization (65-67 Kd). While tumor cells retain the specificity of the intermediate filament, keratin, the individual cells express products of differentiation as measured by keratin expression independently of their cytologic atypia. At the deeper invasive margin of the tumor, the neoplastic cells synthesize keratin proteins in an unpredictable fashion.

Histochemical determination of copper, zinc, and iron in some benign and malignant tissues.

Histochemical tests for copper, iron, and zinc revealed their presence in the stratum germinativum of normal skin and skin from which an early melanoma and basal cell and squamous cell carcinomas originated. However, only copper was seen in invasive cells of basal and squamous cell carcinomas originating from skin. The normal oral mucous membrane was free of copper, iron, and zinc, but cells of invasive squamous carcinoma originating from the oral mucous membrane contained copper. The fluorescent brightening agent, applied as a counterstain, aided in the location of the specimen.