The epidermal basement membrane in basal cell carcinoma: an immunohistochemical study

An immunohistochemical study of basal cell carcinomas of varying histological type, using a panel of antibodies to constituents of the epidermal basement membrane, showed marked deficiencies in the expression of the antigens identified by the antibodies LH7.2, GB3 and G71. There was no correlation between loss of immunoreactivity to these antibodies and the histological features of the tumour.     

Oestrogen and progesterone receptor expression in melasma: an immunohistochemical analysis

Background Melasma is a commonly acquired symmetrical hypermelanosis on sun‐exposed areas of the skin. The development of melasma appears to be associated with increased levels of oestrogen, exposure to sunlight and a genetic predisposition. Several in vitro studies have partially clarified the effects of oestrogen and progesterone on melasma. However, oestrogen receptor (ER) and progesterone receptor (PR) expression in melasma‐affected skin has not been investigated to date, except for one case report on ER expression. Objective The purpose of this study was to compare ER and PR expression between hyperpigmented areas and unaffected areas of facial skin in patients with melasma. Methods Biopsies were performed on skin lesions and adjacent‐unaffected facial skin in 33 Korean women with melasma. The sections were stained using haematoxylin and eosin, Fontana‐Masson, and antibodies to NKI/beteb, ERα, β and PR. Results The immunohistochemical expression of ERβ showed an increasing tendency in epidermal lesions without statistical significance. Expression of PR was significantly increased in the epidermal lesions compared with unaffected skin on the computer‐assisted image analysis. Interestingly, there was increased ERβ expression in the dermal lesions especially around small blood vessels and fibroblast‐like cells compared with unaffected dermis on the semi‐quantitative analysis. However, there was no significant difference in the expression of PR between the dermal lesions and unaffected dermis. Conclusion The results of this study may provide useful information for further investigation into the pathogenesis and therapeutic approaches for treating melasma in relation to hormonal factors. The role of ER in the dermis in association with dermal environment such as blood vessels and fibroblasts remains to be further clarified.     

Clinicopathological and immunohistochemical study of basal cell carcinoma with reference to the features of basement membrane.

One hundred and thirty-five cases of basal cell carcinoma (BCC) were investigated, focusing upon factors determining a postoperative prognosis. Out of the 135, nine tumors on the face recurred. All of these nine tumors were insufficiently extirpated at the initial operation, and showed micronodular or infiltrative patterns with stromal fibrosis. Dividing the degree of dermal invasion into four levels, all tumors with recurrence reached levels 3 and 4, the two deeper groups. The immunohistochemical study using anti-laminin and anti-type IV collagen antibodies showed various changes of staining pattern around tumor cell nests, such as attenuation, disruption, and thickening of basement membrane, in contrast with the normal thinly continuous staining around nontumorous control epidermis. The disruption of basement membranes was remarkable around the tumor cells showing a micronodular growth pattern, although the discontinuity of basement membrane was observed in all types of BCCs to a greater or lesser degree. Ultrastructural thickening, multiplication, or discontinuity of basement membranes were found in all 19 cases examined with a greater or lesser degree, although they were most frequently observed around the cell nests showing micronodular growth patterns. It was concluded that deep dermal and marginal invasions were the most ominous signs of recurrence of BCCs. Although the disruption of basement membranes might participate in the local aggressiveness of BCCs, especially in the tumor cells showing micronodular infiltrative growth, other factors may concern the recurrence of BCCs.     

Involucrin in squamous and basal cell carcinomas of the skin: an immunohistochemical study

Involucrin is a precursor of cross-linked protein of human stratum corneum, and its appearance in the upper layers of the epidermis is a function of the normal differentiation of the keratinocyte. Cases of basal cell and squamous cell carcinoma were evaluated for the presence of involucrin using immunoperoxidase techniques on paraffin sections. Basal cell carcinomas were negative for involucrin with staining restricted to squamous horn cysts, while squamous cell carcinomas stained strongly, particularly in large keratinized cells. Cases of squamous cell carcinoma in situ (Bowen's disease) revealed increased staining for involucrin with staining of dyskeratotic cells at all levels in the epithelium. Abnormal patterns of staining were also noted in non-neoplastic epidermis adjacent to carcinomas. Immunohistochemical staining for involucrin identifying abnormal or premature keratinization is a sensitive marker for dyskeratosis in squamous epithelia and may have applications in the histopathologic evaluation of skin specimens.     

Histochemical and ultrastructural study of cytochrome-oxydase in basal cell epithelioma]

The activity of cytochrome-oxydase is less pronounced in the basal cell epithelioma than in the basal cells of the surrounding normal epidermis. The electron microscopic study of the localisation of the enzyme reveals that the cells of the basal cell epithelioma contain two types of mitochondria, the first of which having the same size as the epidermal mitochondria, and the other being giant mitochondria. The latter seem to be less active. The weak reactivity of the basal cell epithelioma for cytochrome-oxydase is probably due to the reduction of the cristae in mitochondria and to the functional deficiency of the giant mitochondria.     

An immunohistochemical study of laminin in basal cell carcinoma

Background: Laminins are components of the extracellular matrix that mediate cell adhesion, growth, migration, proliferation and differentiation. Basement membrane (BM) laminins, in particular, may play a role in enhancing carcinoma cell motility.
Aim: To evaluate the distribution pattern of laminin in basal cell carcinoma (BCC), as regards the basement membrane, cellular cytoplasm, peritumoral lacunae and surface epithelium and to correlate laminin distribution with different variants of BCC.
Patients and Methods: Skin biopsy specimens were obtained from 21 BCC patients for routine histopathological and immunohistochemical study. Laminin was evaluated qualitatively and semiquantitatively using monoclonal mouse antihuman antibody (Dako-Laminin, 4C7. Code No: MO638, which reacts with the terminal globular domain of the α5 chain)
Results: The majority of BCC cases showed patchy cytoplasmic distribution of laminin. The BM expression of laminin, in most cases, was well defined, fine and linear with irregular areas of thickening. Staining intensity was moderate in differentiated and mixed variants, weak in superficial spreading and absent in morpheic types.
Conclusion: Cytoplasmic and basement membrane laminin is important in the pathogenesis and invasion of BCC. Most laminin was in basement membrane zone (BMZ), and the more differentiated the tumor, the more cytoplasmic and BM staining it expressed.     

Neuroendocrine differentiation in basal cell carcinomas: a retrospective immunohistochemical and ultrastructural study

Recent investigations have suggested that in addition to basaloid cells, basal cell carcinomas (BCC) may also contain Langerhans cells and neuroendocrine cells. In order to establish the relative frequency of neuroendocrine differentiation in BCC, we performed a retrospective study of 50 consecutive BCC using conventional histochemical, immunocytochemical and ultrastructural methods. Argyrophil staining according to Grimelius was used for initial identification. Tumors containing argyrophil cells, as well as randomly selected tumors without these cells, were immunocytochemically stained with a panel of antisera against neurohormonal peptides. Only 2 tumors with argyrophil cells showed occasional somatostatin immunoreactivity; peptide hormone immunoreactivity could not be detected in any of the others. The somatostatin immunoreactive tumors, as well as 3 others, were subjected to electronmicroscopy to study the presence of dense-core secretory granules. However, these characteristic intracytoplasmatic structures could be detected in none. It is concluded that if neuroendocrine differentiation exists at all in BCC, it must be extremely rare. Our results indicate that reliable identification of multiple lines of differentiation in neoplasms can only be performed using combinations of (immuno) histochemical and ultrastructural techniques.     

Extra‐facial melasma: clinical,histopathological, and immunohistochemical case‐control study

Background Extra‐facial melasma is a prevalent dermatosis in some populations with special characteristics in relation to its clinical aspects and probable etiopathogenic factors. Few studies have attempted to address this alteration of pigmentation, which has become a challenge in clinical Dermatology. Objective To assess the clinical histopathological and immunohistochemical characteristics of extra‐facial melasma, comparing affected, and unaffected sites. Methods Case‐control study with 45 patients in each group (melasma and disease‐free volunteers), assessing their clinical characteristics. In 36 patients, biopsies were performed on the lesion and the normal perilesional skin. Specimens were stained with HE and Fontana‐Masson, and melanocytes analysed by immunohistochemistry. Objective measurements were accomplished by a specifically designed image analysis software. Results The melasma group had a mean age ± SD of 56.67 ± 8 years, the majority of them were women (86.7%) and 82.1% of the female cases had reached menopause. There were no significant differences between groups in terms of presence of comorbidities, use of medications or hormone therapies. For extra‐facial melasma patients, family history of this dermatose and of previous facial melasma was significantly higher than in the control group (P < 0.05). The HE staining showed increased rectification and basal hyperpigmentation, solar elastosis, and collagen degeneration in the pigmented area (P < 0.05). There was a significant increase in melanin density in melasma biopsies, but the immunohistochemical tests did not detect a difference between the groups in terms of number of melanocytes. Conclusion Extra‐facial melasma appears to be related to menopause, family history, and personal history of facial melasma, in the studied population. Histopathology revealed a pattern similar to what has been described for facial melasma, with signs of solar degeneration, and a similar number of melanocytes, when comparing patients, and controls, suggesting that the hyperpigmentation is most likely the result of abnormal melanin production or distribution.     

An immunohistochemical study of basal cell carcinoma and trichoepithelioma.

The histologic distinction between tricheopithelioma and basal cell carcinoma may be difficult in small biopsies. Immunohistochemical stains have been used to help make this distinction; however, published studies have generally been limited to a few antibodies. To this end we performed a comprehensive immunohistochemical analysis of 20 basal cell carcinomas and 10 tricheopitheliomas from our files, in search of a consistent pattern of reactivity to distinguish the neoplasms in biopsies. The antibodies used were: low molecular weight keratin (Cam 5.2), Cytokeratin 7, (CK7), Cytokeratin 20, (CK20), Carcino-embryonic antigen (CEA), CD30 (Ki-1), bcl-2, Ham 56, HPCA-I (CD34), and Ulex Europaeus type I. In our study, bcl-2 stained all but one basal cell carcinoma in a diffuse pattern, whereas all tricheopitheliomas showed staining of the outermost epithelial layer. No other stain proved to be an independent marker for either neoplasm and no consistent immunohistochemical profile for either neoplasm emerged. Thus, we conclude that bcl-2 may be of some value in distinguishing basal cell carcinoma from tricheopithelioma, limited by the quantitative nature of the difference in staining. Histologic criteria applied to H&E-stained sections remain the cornerstone of histologic diagnosis.     

Neuroendocrine differentiation in basal cell carcinoma. An immunohistochemical study

Neuroendocrine differentiation in basal cell carcinoma (BCC) of the skin has been reported in the past on the basis of ultrastructural findings, argyrophilia, and the immunohistochemical detection of neuropeptides in such neoplasms. To assess further the relative frequency of neuroendocrine differentiation in BCC, paraffin sections of 53 randomly chosen cases were evaluated for Churukian-Schenk stain positivity and the expression of sensitive neuroendocrine markers. These included neuron-specific enolase (using a highly absorbed monospecific antiserum), chromogranin-A, synaptophysin, neurofilament protein, and Leu-7 antigen. Although 25% of cases demonstrated argyrophilia with the Churukian-Schenk method, suggesting a high frequency of possible neuroendocrine differentiation, immunohistochemical evidence of the same was observed in only two tumors (4%). These demonstrated immunoreactivity for chromogranin and neuron-specific enolase. The results of this analysis suggest that neuroendocrine differentiation in BCC is relatively uncommon, and that it is not reliably predicted by the results of argyrophil stains done on paraffin sections.     

The basement membrane zone in lichen sclerosus: an immunohistochemical study

The alteration in expression of basement membrane zone (BMZ) components in lichen sclerosus was investigated by immunohistochemical staining of skin biopsies from seven patients with histologically confirmed disease compared with controls. Monoclonal antibodies and polyclonal sera directed against proteins of the hemidesmosomes, anchoring fibrils, lamina lucida, lamina densa and BMZ collagens were used. Characteristic histological appearances at the dermo-epidermal junction were reflected in widespread alterations in antigen expression in the epidermal basement membrane and the papillary dermis. Expression of the proteins which constitute the structural scaffold (collagen IV and VII) were increased in lichen sclerosus. Expression of hemidesmosomal proteins which mediate adhesion and cell to matrix interaction (α6/β4 and hullous pemphigoid antigen) and expression of anchoring filament components were markedly reduced, suggesting that the epidermal cells are exposed to selective damage.     

Light microscopic, immunohistochemical, and ultrastructural alterations in patients with melasma

Despite new technologies, few studies have assessed the histologic alterations in patients with melasma. Using current technologies, the present study was designed to re-evaluate the light microscopic, immunohistochemical, and ultrastructural changes of the hyperpigmented and adjacent normal skin of patients with melasma. Twenty-one patients were included in this study. Two millimeter punch biopsies were taken from the hyperpigmented and adjacent normal skin of the face. The integrity of the epidermis and dermis was assessed by light microscopy, computer-assisted image analysis, immunohistochemistry, and electron microscopy. Stains included hematoxylin-eosin and Fontana-Masson for melanin detection. Immunostaining was performed using Mel-5 antibody and CD1a antibody as markers for melanin and Langerhans cells, respectively. However, mild lymphohistiocytic infiltrates were present in 75% of the hyperpigmented areas. The areas of hyperpigmentation showed increased deposition of melanin in the epidermis and dermis of all cases. There was a statistically significant increase in the content of epidermal melanin. There were no quantitative increases in melanocytes in the hyperpigmented areas of skin. However, the melanocytes in the hyperpigmented areas were larger, intensely stained cells with very prominent dendrites. Electron microscopy revealed more melanosomes in keratinocytes, melanocytes, and dendrites in the involved skin in comparison to the uninvolved skin. The results of this study suggest that melasma is a consequence of specific hyperfunctional melanocytes that cause excessive melanin deposition in the epidermis and dermis.     

Superoxide dismutase in psoriasis, squamous cell carcinoma and basal cell epithelioma: an immunohistochemical study

Monoclonal antibodies against human Cu,Zn-superoxide dismutase (SOD) and Mn-SOD were used to stain frozen sections of normal and abnormal human skin. In normal human epidermis, the Cu,Zn-SOD antibody almost exclusively stained the basal cells. Mn-SOD antibody weakly stained the whole of the epidermis but more predominantly the basal cell layer. In psoriasis, Cu,Zn-SOD antibody mainly stained the basal cells of the lowest parts of the elongated rete ridges. Basal cells corresponding to the tip of the dermal papillae were weakly stained. Mn-SOD staining was considerably decreased in the psoriatic epidermis. In squamous cell carcinoma, staining with both Cu,Zn-SOD and Mn-SOD antibodies was decreased, and single cells positive for Cu,Zn-SOD were scattered throughout the tumour nests. In basal cell epithelioma, Cu,Zn-SOD staining was intense and diffusely distributed throughout the tumour nests, while Mn-SOD staining was absent.     

Regression in basal cell carcinoma: an immunohistochemical analysis

Summary Spontaneous regression of some cutaneous tumours is well recognized, and is thought to result from an immunological response to the tumour, Regression has previously been noted in basal cell carcinomas, but no studies defining the role of the immune response in the regression of this malignancy have been performed. We have examined 45 primary basal cell carcinomas (BCCs) (20 nodular, 25 superficial) and identified the cellular phenotypes and activation states of the cells infiltrating primary regressing and non-regressing BCCs, by immunocytochemistry. We have found a significantly increased number of CD3⁺ and CD4⁺ T cells infiltrating regressing compared with non-regressing tumours, and the expression of interleukin-2 receptor (an early activation marker for T cells) was also increased. There were no significant differences in class II major histocompatibility complex (MHC), CD1, or macrophage antigen expression in these groups. These findings suggest that activated CD4⁺ cytokine-secreting cells are important in the regression of BCCs.     

Cytokeratins as markers of follicular differentiation: an immunohistochemical study of trichoblastoma and basal cell carcinoma.

Trichoblastoma(s) (TB) are benign neoplasms of follicular differentiation frequently found in nevus sebaceus. Many morphologic features are shared with nodular basal cell carcinoma(s) (BCC), sometimes rendering the differential diagnosis difficult. Because both neoplasms can simulate components of mature hair follicles histologically, we attempted to corroborate this by immunohistochemical examination of cytokeratins and hair keratins differentially expressed in the hair follicle. Trichoblastoma(s) and BCC showed homogenous expression of CK14 and CK17. The innermost cells of the tumor nodules in all TB and in 72% of BCC were positive for CK6hf. Using a specific CK15 antibody, 38% of TB showed a focal labeling and all BCC remained negative; 70% of TB and 22% of BCC expressed CK19. CK8 was expressed by numerous Merkel cells present in all TB but in none of the BCC examined. All type I and II hair keratins tested, (especially hHa1, hHa5, and hHa8) remained negative in all tumors examined. Trichoblastoma(s) and BCC show consistent expression of CK6hf, CK14, and CK17; variable expression of CK15 and CK19; and absence of hair keratins. This indicates a differentiation toward the outer root sheath epithelium or the companion layer and not toward the inner root sheath, matrix, or cortex.