Distribution of extracellular and cytoplasmic domains of the alpha 3 and alpha 6 integrin subunits in solid tumors.

Integrins play an important role in malignant transformation and the invasion of tumors. They mediate cell-cell and cell-matrix interactions and participate in transduction of signals across the plasma membrane, processes dependent on the extracellular and cytoplasmic domains of integrins. We studied a selection of solid tumors by immunohistochemistry using monoclonal antibodies (MAbs) against the extracellular domain and the cytoplasmic variants (A and B) of the alpha 3 and alpha 6 integrin subunits. The tissue-specific expression of ecto- and cyto-domains of alpha 3 and alpha 6 is maintained in a subset of breast, colon, kidney and parotid tumors. In a few breast tumors, there was a switch in variant expression in that alpha 6B was detected instead of alpha 6A in normal breast tissue. In many colon and parotid tumors, one of the variants of alpha 6 was missing, while both were detectable in the corresponding normal tissues. In contrast, coexpression of the alpha 6 variants was found in some kidney tumors, whereas only one of the variants was detected in the normal tissue. In a minority of colon and kidney tumors, the cyto-domains of alpha 3 and alpha 6 were undetectable and total absence of alpha 3 and alpha 6 was noted in a subset of breast, colon, kidney and parotid tumors. These observations show that expression of the integrin variants in tumors varies considerably and support the concept that changes in expression may contribute to malignant behavior.

Cytokeratin subtypes and involucrin in squamous cell carcinoma of the vulva. An immunohistochemical study of 41 cases.

BACKGROUND: Histologic grade seems to be of limited prognostic significance in patients with vulvar carcinoma. However, the study of cytokeratin expression is of potential interest because it allows a more precise evaluation of the degree of squamous differentiation. This study was conducted to investigate whether differences in cytokeratin expression exist between normal vulvar epithelium and vulvar carcinoma and whether these differences are prognostically significant. METHODS: The expression of several differentiation markers, i.e., cytokeratin (CK) 10, CK 13, and involucrin, was studied in samples of 41 vulvar carcinomas. The expression of CK 8, 10, 13, and 14 was compared with CK expression in normal vulvar epithelium and was correlated with tumor grade and tumor growth pattern. Tumor growth pattern was considered type A if infiltrating tumor cell nests showed a layer of small, basaloid cells bordering the surrounding mesenchymal tissue and was considered type B if this was not the case. Prognosis was based on whether disease recurred or not. RESULTS: Sixteen patients had disease recurrence. No prognostic significance of tumor grade was found. Tumor growth pattern was prognostically significant: in patients with a type A tumor, recurrence was observed less often than in patients with a type B tumor (P = 0.03). Cytokeratin 14, typical for basal cells of normal vulvar epithelium, was expressed in all tumors, whereas CK 8 was not expressed in any tumor. A relationship between tumor growth pattern and the concordant expression of differentiation markers was observed: in 55% of type A tumors and in none of type B tumors, concordant expression of CK 10, CK 13, and involucrin was found. CONCLUSION: The expression of differentiation markers in vulvar carcinoma is related strongly to the tumor growth pattern, and this pattern is prognostically significant.

Modulation of mammary carcinoma cell phenotype and keratin expression patterns by retinoic acid.

Immunohistochemical and biochemical procedures were used to study the influence of retinoic acid (RA) on cellular expression and distribution of cytokeratins (CKs) in feline mammary carcinoma cells. These cells were grown in vitro as established cell lines (K248C and K266) and in vivo as xenografts in athymic mice. The results were compared with the distribution of CKs in normal feline mammary gland and in a series of invasive mammary carcinomas previously probed with a panel of monoclonal antibodies specific for individual CKs. Coexpression of CKs of both major mammary gland cell types (myoepithelial cells, MECs, CKs 5/14 positive, and luminal epithelial cells, LECs CKs8/18 positive) by K248C and K266 cells, suggested a stem cell-like character of both cell lines. RA increased CK19 expression in both cell lines and CK19 was also present in tumors developed in nude mice from both RA untreated (CK19 negative) and RA-treated (CK19 positive) K248C and K266 cells. In addition, RA had cell line specific effects as well. RA treatment induced differentiation of K248C cells to more mature LEC-like cells and this change was accompanied by the loss of the MEC keratins CKs 5/14. Under the same culture conditions however, RA treatment did not induce morphological changes in the K266 cell line and the expression of CKs 5/14 was not significantly reduced. These findings suggest that the modulation of CK19 and CKs 5/14 expression observed in mammary carcinoma cells upon RA treatment might be regulated through different pathways.

Cytokeratins as markers of initial stages of squamous metaplasia in feline mammary carcinomas.

Expression of keratins (cytokeratins, CK) known to be suitable markers for different types of epithelial differentiation was analyzed in specimens of feline mammary tissue. A panel of specific anti-CK monoclonal antibodies (MAb) was used to determine CK distribution pattern in normal feline tissues (n = 3), and in benign (n = 18) and malignant (n = 20) feline mammary tumors. In selected tumors, the CK distribution pattern was also determined by biochemical methods. A MAb specific for alpha-smooth muscle actin was used to discriminate between myoepithelial cells and luminal epithelial cells. In normal mammary gland tissues, 6 MAb reacted exclusively, either with myoepithelial cells or with luminal epithelial cells. Luminal epithelial cells reacted with MAb specific for CK typical of simple epithelia, whereas myoepithelial cells reacted with MAb specific for CK in basal cells of stratified epithelia. A similar distribution of CK was detected in specimens from benign tumors, except that CK4 was not detected in normal mammary gland tissues and was detected in some ducts in specimens with adenosis. Almost all tumor cells in specimens from malignant tumors reacted with MAb specific for CK typical of simple epithelia. Concomitant expression of CK typical of stratified epithelia was detected in small or large subpopulations of tumor cells in 70% of carcinomas. Cytokeratins typical of basal cell layers and typical for suprabasal layers of inner stratified epithelia were detected. Cytokeratins typical of stratified epithelia were always found in areas of squamous metaplasia, but also were found in adenocarcinomal cells surrounding these areas.(ABSTRACT TRUNCATED AT 250 WORDS)

Patterns of expression of feline cytokeratins in healthy epithelia and mammary carcinoma cells.

Expression of keratins (cytokeratins, CK) in healthy feline epithelia and 2 established feline mammary carcinoma cell lines was examined immunohistochemically and by use of immunoblotting analysis. A panel of specific anti-CK monoclonal antibodies (MAb) identifying epitopes unique to individual keratins or shared by 2 (or 3) CK polypeptides was used. Besides already available anti-human CK MAb, this panel of MAb consisted of 9 newly generated anti-human CK MAb and 1 newly generated anti-feline CK MAb. Immunohistochemical analysis on normal epithelia revealed that most of the anti-human CK MAb and the anti-feline CK MAb reacted with both feline and human epithelia, with a comparable tissue distribution pattern. However, slight differences in CK tissue distribution pattern between human beings and cats were detected by one MAb. Immunoblotting analysis revealed that all anti-human CK MAb that were immunohistochemically reactive with feline tissues detected analogous CK in cats, indicating the presence of a number of common epitopes on human and feline CK. Two continuous cell lines derived from 2 distinct feline mammary adenocarcinomas, K248C and K266, were analyzed with respect to their CK phenotype. Although no difference in CK expression between the 2 cell lines was detected in vitro, a difference in CK phenotype was detected on subcutaneous transplantation of the 2 cell lines into nude mice. Although the K248C-induced adenocarcinomas maintained the same CK phenotype as observed in vitro, the CK pattern of the K266 heterotransplants, growing as adenosquamous carcinomas, changed with squamous differentiation. Our findings confirm the high degree of homology between mammalian CK, and on the basis of those findings, we suggest that CK proteins provide a set of markers valuable for the characterization of normal and neoplastic feline tissues and for studies of squamous metaplasia.

Keratin subtypes in carcinomas of the uterine cervix: implications for histogenesis and differential diagnosis.

Normal epithelia and carcinomas of the human uterine cervix were studied by monoclonal antibodies chain specific for cytokeratins 4, 8, 10, 13, 14, 18, and 19. Most cells in 13 examined squamous carcinomas revealed a cytokeratin phenotype detected in ectocervical basal cells and endocervical subcolumnar reserve cells: 8+, 14+, 18+, 19+, 4-, 10-, 13-. We propose that these two cell types are closely related or identical and that squamous carcinoma of the cervix originates in this cell type. In more differentiated tumor cells cytokeratins 4, 10, and 13, which are present in suprabasal layers of the normal ectocervical epithelium, were coexpressed with basal cell cytokeratins. Thus, contrary to previous beliefs, all cytokeratins detected in carcinomas were also present in normal epithelium of uterine cervix. The cytokeratin profile of cervical adenocarcinomas corresponded to that of columnar endocervical cells (8+, 18+, 19+), although two of the three adenocarcinomas also expressed cytokeratin 4, which in the normal endocervix was detected in scattered single columnar cells only. The new monoclonal antibody DE-K14, specific for cytokeratin 14, proved a specific marker of subcolumnar reserve cells in the endocervix. It was also the only one that reacted with all cervical squamous carcinomas but with none of the cervical adenocarcinomas and, as such, has a potential value for pathological differential diagnosis of cervical tumors.