Expression of GATA3 and Cytokeratin 14 in Urinary Bladder Carcinoma (Histopathological and Immunohistochemical Study)

BACKGROUND: Urothelial carcinoma (UC) with squamous differentiation (SD) is the most common histologic variant of bladder carcinoma and its presence is associated with poor prognosis which may need early radical cystectomy to avoid progression and recurrence. It is difficult to detect few foci of SD, especially nonkeratinizing or early switch from urothelial to squamous epithelium on only morphological basis. Combination of GATA3 and Cytokeratin 14 (CK14) could be helpful in differentiating pure UC, UC with SD and pure squamous cell carcinoma (SCC). AIM: Assessment of GATA3 and CK14 expression in urinary bladder carcinoma and correlation with clinical and histopathological variables, for both diagnostic and prognostic purposes. MATERIALS AND METHODS: Sixty cases of archived paraffin blocks of urinary bladder carcinoma were tested for GATA3 and CK14 expression by immunohistochemistry using a rabbit monoclonal antibody against human CK 14 and mouse monoclonal antibody against GATA3, respectively. RESULTS: There is a significant correlation between GATA3 immunohistochemical expression and histological tumor subtypes of bladder carcinoma (p < 0.001), i.e. the GATA3 is a useful marker for urothelial origin especially in papillary UC. There is a significant correlation between GATA3 immunohistochemical expression and UC grade (p < 0.001). CK14 showed positive cytoplasmic staining in 9/14 (64.3%) cases of UC with SD and (13/13) (100%) cases of pure SCC and negative in 33/33(100%) cases of UC other than UC with SD. CK14 had sensitivity (64.3%) and specificity (100%) for areas of SD. CONCLUSION: GATA3 is a specific immunohistochemical marker for urothelial origin. CK14 is a highly specific and sensitive immunohistochemical marker of squamous cell carcinoma.

The Combined Thermoresponsive Cell-Imprinted Substrate, Induced Differentiation, and "KLC Sheet" Formation

Purpose: Stem cells can exhibit restorative effects with the commitment to functional cells. Cell-imprinted topographies provide adaptable templates and certain dimensions for the differentiation and bioactivity of stem cells. Cell sheet technology using the thermo-responsive polymers detaches the "cell sheets" easier with less destructive effects on the extracellular matrix (ECM). Here, we aim to dictate keratinocyte-like differentiation of mesenchymal stem cells by using combined cell imprinting and sheet technology. Methods: We developed the poly dimethyl siloxane (PDMS) substrate having keratinocyte cell-imprinted topography grafted with the PNIPAAm polymer. Adipose tissue-derived mesenchymal stem cells (AT-MSCs) were cultured on PDMS substrate for 14 days and keratinocyte-like differentiation monitored via the expression of involucrin, P63, and cytokeratin 14. Results: Data showed the efficiency of the current protocol in the fabrication of PDMS molds. The culture of AT-MSCs induced typical keratinocyte morphology and up-regulated the expression of cytokeratin-14, Involucrin, and P63 compared to AT-MSCs cultured on the plastic surface (p<0.05). Besides, KLC sheets were generated once slight changes occur in the environment temperature. Conclusion: These data showed the hypothesis that keratinocyte cell imprinted substrate can orient AT-MSCs toward KLCs by providing a specific niche and topography.

Primary ectocervical epithelial cells display lower permissivity to Chlamydia trachomatis than HeLa cells and a globally higher pro-inflammatory profile

The tumoral origin and extensive passaging of HeLa cells, a most commonly used cervical epithelial cell line, raise concerns on their suitability to study the cell responses to infection. The present study was designed to isolate primary epithelial cells from human ectocervix explants and characterize their susceptibility to C. trachomatis infection. We achieved a high purity of isolation, assessed by the expression of E-cadherin and cytokeratin 14. The infectious progeny in these primary epithelial cells was lower than in HeLa cells. We showed that the difference in culture medium, and the addition of serum in HeLa cultures, accounted for a large part of these differences. However, all things considered the primary ectocervical epithelial cells remained less permissive than HeLa cells to C. trachomatis serovar L2 or D development. Finally, the basal level of transcription of genes coding for pro-inflammatory cytokines was globally higher in primary epithelial cells than in HeLa cells. Transcription of several pro-inflammatory genes was further induced by infection with C. trachomatis serovar L2 or serovar D. In conclusion, primary epithelial cells have a strong capacity to mount an inflammatory response to Chlamydia infection. Our simplified purification protocol from human explants should facilitate future studies to understand the contribution of this response to limiting the spread of the pathogen to the upper female genital tract.

Sebaceoma of a Meibomian Gland of the Upper Eyelid

Over a period of 1 year, a 74-year-old man slowly developed a painless left upper eyelid intratarsal mass. The skin was movable over the lesion. At surgery, a well-circumscribed, yellow-white, partially cystic tumor was encountered. Histopathologically it was composed of a random mixture of basaloid and sebaceous cells arranged in interconnecting cords. Immunohistochemical evaluation disclosed epithelial membrane antigen, adipophilin, and cytokeratin 14 positivity. These findings led to the diagnosis of a sebaceoma. The tumor cells abnormally failed to express mismatch repair proteins for MLH1 and PMS2. The patient did not have a personal history of any visceral malignancy, but his father had died at the age of 46 years and a daughter at the age of 33 years from colonic carcinomas. The implications of this periocular sebaceoma for the Muir-Torre syndrome are explored.

First description of basaloid carcinoma of the canine mammary gland: case report

ABSTRACT The objective of this case report was to describe histopathological and immunohistochemical characteristics of the first reported basaloid carcinomas in the canine mammary gland. Two bitches were treated for tumors in the mammary gland and underwent mastectomy. Microscopic evaluation of these tumors revealed epithelial cells arranged in a predominantly solid pattern with hyperchromatic peripheral cells arranged in a palisade pattern. Metastases in regional lymph nodes were found in both animals, and one bitch exhibited pulmonary metastasis. Immunohistochemistry revealed positive labeling for the basal cell markers cytokeratin 14 and p63. Histopathological and immunohistochemical findings led to diagnoses of basaloid carcinoma of the canine mammary gland with regional and distant metastasis.

CYTOKERATIN PROFILES OF CANINE ANAL SAC AND HEPATOID GLAND NEOPLASMS

Ten adenocarcinomas of the apocrine gland of the anal sac and 11 hepatoid gland neoplasms were studied to determine the coordinate expression of cytokeratin 7 (CK7) and cytokeratin 14 (CK14) using commercially available antibodies. Hepatoid gland neoplasms included hepatoid gland adenomas, carcinomas and a single epithelioma. All 10 (100%) adenocarcinomas of the apocrine gland of the anal sac had CK7[Formula: see text]/CK14[Formula: see text] immunophenotype, whereas all 11 (100%) of hepatoid neoplasms expressed CK7[Formula: see text]/CK14[Formula: see text] immunophenotype. Hepatoid gland adenomas, carcinomas, and epithelioma could not be differentiated based on the cytokeratin immunophenotype. Hepatoid gland neoplasms could be differentiated from adenocarcinomas of the apocrine gland of the anal sac by differences in the CK7/CK14 immunophenotypes with a [Formula: see text]-[Formula: see text]. The results of this study provide further support for the use of coordinate expression of CK7/CK14 to differentiate apocrine gland adenocarcinomas of the anal sac (CK7[Formula: see text]/CK14[Formula: see text]) from hepatoid gland neoplasms (CK7[Formula: see text]/CK14[Formula: see text]).

MAPK pathway involved in epidermal terminal differentiation of normal human epidermal keratinocytes

ObjectiveTo investigate the effect of mitogen-activated protein kinase (MAPK) signaling pathway in epidermal terminal differentiation.MethodsThe MAPK pathways (p38, ERK1/2, JNK) were inhibited by SB203580, PD98059, and SP600125 in normal human epidermal keratinocytes (NHEKs), respectively. Western blotting assays were performed to detect expression of filaggrin and differentiation-related proteins. The mRNA expressions of differentiation-related proteins were detected by real-time quantitative PCR (qRT-PCR).ResultsInhibition of MAPK pathway by SB203580, PD98059, and SP600125 resulted in significant reduction of filaggrin expression in NHEKs. Inhibition of the p38 MAPK pathway decreased the expression of differentiation-related proteins (cytokeratin 5, cytokeratin 14, ST14, and SPRR3), Akt, and NF-κB. Inhibition of JNK also suppressed expression of cytokeratin 14, SPRR3, Akt, and NF-κB. However, inhibition of ERK1/2 merely decreased expression of SPRR3 and Akt.ConclusionMAPK pathways regulates epidermal terminal differentiation in NHEKs. The p38 signaling pathway plays an especially important role.