RESUMEN

Recent studies have demonstrated the role of caspase-14 in terminally differentiated keratinocytes, and its expression may decrease the magnitude of tumors in the epidermis. In the present study, we assessed the potential of luteolin (LUT) to elicit the expression of caspase-14 in terminal differentiation of human keratinocytes. The semi-qualitative RT-PCR data revealed a significant level of caspase-14 expression in LUT-treated human immortalized keratinocytes (HaCaT) with respect to untreated cells. The quantitative data (ELISA) further supported the potency of LUT to induce caspase-14 expression at 3.19 ng/ml when compared to 1.29 ng/ml of vitamin D3 (positive control). Further, the enhanced expression of human involucrin gene in LUT-treated HaCaT cells confirmed its ability to drive terminal differentiation in these cells. These preliminary results provide first-hand information about the in vitro potential of LUT to elicit the expression of caspase-14, thereby inducing terminal differentiation in human keratinocytes.

Asunto(s)

Caspasa 14/biosíntesis , Diferenciación Celular/efectos de los fármacos , Células Epidérmicas , Queratinocitos/citología , Luteolina/farmacología , Caspasa 14/metabolismo , Línea Celular , Colecalciferol/farmacología , Humanos , Precursores de Proteínas/biosíntesis

RESUMEN

OBJECTIVES: Caspase 14 is reduced in adenocarcinomas of the stomach and colon. In contrast, breast and lung adenocarcinomas frequently show an overexpression of caspase 14. Salivary gland adenocarcinomas have not been evaluated for potential aberrant caspase 14 expression. MATERIALS AND METHODS: Samples from salivary gland carcinomas (n = 43) were analysed by immunohistochemistry (caspase 14, filaggrin, GATA3 and Ki67) and fluorescence in situ hybridization. RESULTS: Caspase 14 is not expressed in normal salivary glands, while in a subfraction of carcinomas (32%) an aberrant expression was found. Filaggrin could not be detected. Caspase 14 staining was not associated with tumour dedifferentiation, GATA3 expression or amplification of gene locus 19p13. CONCLUSION: In summary, aberrant expression of caspase 14 can be found in a subfraction of salivary gland carcinomas but could not be used as a biomarker for a specific carcinoma subtype of the salivary gland.

Asunto(s)

Carcinoma de Células Escamosas/enzimología , Caspasa 14/biosíntesis , Neoplasias de Cabeza y Cuello/enzimología , Neoplasias de las Glándulas Salivales/enzimología , Anciano , Anciano de 80 o más Años , Biomarcadores de Tumor/biosíntesis , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/patología , Caspasa 14/genética , Femenino , Proteínas Filagrina , Factor de Transcripción GATA3/biosíntesis , Amplificación de Genes , Sitios Genéticos , Neoplasias de Cabeza y Cuello/genética , Neoplasias de Cabeza y Cuello/patología , Humanos , Inmunohistoquímica , Hibridación Fluorescente in Situ/métodos , Proteínas de Filamentos Intermediarios/biosíntesis , Antígeno Ki-67/biosíntesis , Masculino , Persona de Mediana Edad , Neoplasias de las Glándulas Salivales/genética , Neoplasias de las Glándulas Salivales/patología , Carcinoma de Células Escamosas de Cabeza y Cuello

RESUMEN

OBJECTIVE: The aim of this study is to determine the expression of caspase-14, a key protein in maturation of squamous epithelia, in archival malignant and premalignant vulvar squamous lesions and examine in-vitro effects of a black raspberry extract (BRB-E) on a vulvar squamous cell carcinoma (VSCC) cell line. METHODS: VSCC cell cultures were exposed to different BRB-E concentrations and used to create cell blocks. Immunohistochemistry for caspase-14 was performed on cell block sections, whole tissue sections, and a tissue microarray consisting of normal vulvar skin, lichen sclerosus (LS), classic and differentiated vulvar intraepithelial neoplasia (cVIN and dVIN respectively), and VSCC. RESULTS: LS demonstrated abnormal full thickness (5/11) or absent (1/11) caspase-14 staining. dVIN often showed markedly reduced expression (4/7), and cVIN occasionally demonstrated either absent or reduced caspase-14 (6/22). VSCC predominantly had absent or markedly reduced caspase-14 (26/28). VSCC cell cultures demonstrated a significant increase in caspase-14 (p=0.013) after BRB-E treatment: 7.3% (±2.0%) of untreated cells showed caspase-14 positivity, while 21.3% (±8.9%), 21.7% (±4.8%), and 22.6% (±5.3%) of cells were positive for caspase-14 after treatment with 200, 400, and 800 µg/mL BRB-E, respectively. Pair-wise comparisons between the treatment groups and the control demonstrated significant differences between no treatment with BRB-E and each of these treatment concentrations (Dunnett's adjusted p-values: 0.024, 0.021, and 0.014, respectively). CONCLUSIONS: Caspase-14 is frequently decreased in premalignant and malignant vulvar squamous lesions, and is upregulated in VSCC cell culture by BRB-E. BRB-E should be further explored and may ultimately be incorporated in topical preparations.

Asunto(s)

Carcinoma de Células Escamosas/tratamiento farmacológico , Carcinoma de Células Escamosas/enzimología , Caspasa 14/biosíntesis , Extractos Vegetales/uso terapéutico , Rubus/química , Neoplasias de la Vulva/tratamiento farmacológico , Neoplasias de la Vulva/enzimología , Carcinoma de Células Escamosas/patología , Línea Celular Tumoral , Femenino , Frutas/química , Humanos , Inmunohistoquímica , Neoplasias de la Vulva/patología

RESUMEN

Caspase-14 is a cysteinyl-aspartate-specific proteinase that is specifically expressed in epidermal keratinocytes. Dysregulation of caspase-14 expression is implicated in impaired skin barrier formation. To elucidate the regulation of caspase-14 in differentiated keratinocytes, we characterized the expression of caspase-14 in normal human epidermal keratinocytes (NHEKs) and two types of three-dimensional (3D) human epidermis culture models, EPI-200 and EPI-201, via RT-PCR and immunoblot analyses. Caspase-14 expression was absent in subconfluent NHEKs, but was present in confluent NHEKs as well as those induced to differentiate by calcium. Caspase-14 expression levels in the 3D epidermis models were almost equal to that in the Ca(2+)-treated differentiated NHEKs. Despite the presence of caspase-14 expression in these models, caspase-14 activity was found only in the mature 3D skin model, EPI-200. This was confirmed by detection of a 17 kDa cleaved fragment of caspase-14 present only in the EPI-200 model. Since glucocorticoid (GC) receptor is required for skin barrier competence, we investigated whether the GC dexamethasone (Dex) and various natural components of common skin moisturizers affect caspase-14 expression in keratinocytes. Dex decreased caspase-14 expression in undifferentiated, but not differentiated, NHEKs. Conversely, Dex increased caspase-14 expression in both 3D skin models, although it did not alter caspase protease activity. Similar to treatment with Dex, treatment of the premature 3D skin mode, EPI-201 with a Galactomyces ferment filtrate markedly increased expression of caspase-14. Further, these results suggest that the effect of Dex, or lack thereof, on caspase-14 expression is dependent on the stage of keratinocyte differentiation.

Asunto(s)

Caspasa 14/metabolismo , Dexametasona/farmacología , Epidermis/metabolismo , Queratinocitos/metabolismo , Productos Biológicos/farmacología , Caspasa 14/biosíntesis , Caspasa 14/genética , Línea Celular , Cosméticos/farmacología , Epidermis/efectos de los fármacos , Humanos , Queratinocitos/efectos de los fármacos , ARN Mensajero/biosíntesis , Receptores de Glucocorticoides/efectos de los fármacos

RESUMEN

Understanding how oral administration of aroma terpenes can prevent sunburn or skin cancer in mice could lead to more effective and safer ways of blocking sun damage to human skin. To establish sunburn preventive activity, female Skh-1 mice were given oral ß-damascenone followed by irradiation with UVR from fluorescent 'sunlamps'. The following endpoints were evaluated versus controls at various times between 1 and 12 days after the terpene: whole genome gene expression and in situ immunohistochemistry of PCNA, keratin 10, filaggrin and caspase 14, and sunburn was evaluated at 5 days. UVR-induced sunburn was prevented by a single oral ß-damascenone dose as low as 20 µL (0.95 mg/g body weight). Microarray analysis showed sunburn prevention doses of ß-damascenone up-regulated several types of cornification genes, including keratins 1 and 10, filaggrin, caspase 14, loricrin, hornerin and 6 late cornified envelope genes. Immunohistochemical studies of PCNA labeling showed that ß-damascenone increased the proliferation rates of the following cell types: epidermal basal cells, follicular outer root sheath cells and sebaceous gland cells. Keratin 10 was not affected by ß-damascenone in epidermis, and filaggrin and caspase 14 were increased in enlarged sebaceous glands. The thickness of the cornified envelope plus sebum layer nearly doubled within 1 day after administration of the ß-damascenone and remained at or above double thickness for at least 12 days. ß-Damascenone protected against sunburn by activating a sebaceous gland-based pathway that fortified and thickened the cornified envelope plus sebum layer in a way that previously has been observed to occur only in keratinocytes.

Asunto(s)

Epidermis/efectos de los fármacos , Epidermis/metabolismo , Proteínas de Filamentos Intermediarios/biosíntesis , Norisoprenoides/farmacología , Quemadura Solar/prevención & control , Administración Oral , Animales , Caspasa 14/biosíntesis , Caspasa 14/genética , Caspasa 14/metabolismo , Proliferación Celular/efectos de los fármacos , Células Epidérmicas , Femenino , Proteínas Filagrina , Inmunohistoquímica , Proteínas de Filamentos Intermediarios/genética , Proteínas de Filamentos Intermediarios/metabolismo , Queratina-10/biosíntesis , Queratina-10/genética , Queratina-10/metabolismo , Ratones , Norisoprenoides/administración & dosificación , Análisis de Secuencia por Matrices de Oligonucleótidos , Antígeno Nuclear de Célula en Proliferación/biosíntesis , Antígeno Nuclear de Célula en Proliferación/genética , Antígeno Nuclear de Célula en Proliferación/metabolismo , ARN Mensajero/química , ARN Mensajero/genética

RESUMEN

The transcription factor Gata-3 is a definitive marker of luminal breast cancers and a key regulator of mammary morphogenesis. Here we have explored a role for Gata-3 in tumor initiation and the underlying cellular mechanisms using a mouse model of "luminal-like" cancer. Loss of a single Gata-3 allele markedly accelerated tumor progression in mice carrying the mouse mammary tumor virus promoter-driven polyomavirus middle T antigen (MMTV-PyMT mice), while overexpression of Gata-3 curtailed tumorigenesis. Through the identification of two distinct luminal progenitor cells in the mammary gland, we demonstrate that Gata-3 haplo-insufficiency increases the tumor-initiating capacity of these progenitors but not the stem cell-enriched population. Overexpression of a conditional Gata-3 transgene in the PyMT model promoted cellular differentiation and led to reduced tumor-initiating capacity as well as diminished angiogenesis. Transcript profiling studies identified caspase-14 as a novel downstream target of Gata-3, in keeping with its roles in differentiation and tumorigenesis. A strong association was evident between GATA-3 and caspase-14 expression in preinvasive ductal carcinoma in situ samples, where GATA-3 also displayed prognostic significance. Overall, these studies identify GATA-3 as an important regulator of tumor initiation through its ability to promote the differentiation of committed luminal progenitor cells.

Asunto(s)

Neoplasias de la Mama/metabolismo , Carcinoma Intraductal no Infiltrante/metabolismo , Caspasa 14/metabolismo , Factor de Transcripción GATA3/metabolismo , Glándulas Mamarias Animales/citología , Glándulas Mamarias Humanas/citología , Neoplasias Mamarias Experimentales/metabolismo , Adulto , Anciano , Animales , Antígenos Transformadores de Poliomavirus/genética , Neoplasias de la Mama/genética , Neoplasias de la Mama/patología , Carcinoma Intraductal no Infiltrante/genética , Carcinoma Intraductal no Infiltrante/patología , Caspasa 14/biosíntesis , Diferenciación Celular , Transformación Celular Neoplásica/genética , Femenino , Factor de Transcripción GATA3/biosíntesis , Factor de Transcripción GATA3/genética , Humanos , Glándulas Mamarias Animales/metabolismo , Glándulas Mamarias Humanas/metabolismo , Neoplasias Mamarias Experimentales/genética , Neoplasias Mamarias Experimentales/patología , Virus del Tumor Mamario del Ratón/genética , Ratones , Persona de Mediana Edad , Células Madre

RESUMEN

Using apoptosis-inducing factor protein as bait in a yeast hybrid assay to screen protein libraries, we identified three proteins that interacted with apoptosis-inducing factor: human homolog of yeast Rad23 protein A (hHR23A), microsomal glutathione S-transferase and caspase-14 (casp-14). In this study, we investigated the expression and function of casp-14 in lung adenocarcinomas (LADC). Our results showed that monoclonal antibodies were specific to casp-14, and that casp-14 was highly expressed in LADC. Casp-14 overexpression correlated with tumor stage, cell differentiation and lymphovascular involvement, suggesting that casp-14 was associated with tumor cell growth and metastatic potential. In vitro, casp-14 interacted with apoptosis-inducing factor, and silencing of casp-14 expression reduced cisplatin resistance. Our data suggest that casp-14 is an anti-apoptotic protein targeting apoptosis-inducing factor and increases cisplatin resistance in LADC cells.

Asunto(s)

Adenocarcinoma/metabolismo , Factor Inductor de la Apoptosis/metabolismo , Proteínas Reguladoras de la Apoptosis/metabolismo , Caspasa 14/biosíntesis , Neoplasias Pulmonares/metabolismo , Adenocarcinoma/enzimología , Adenocarcinoma/patología , Adenocarcinoma del Pulmón , Secuencia de Aminoácidos , Animales , Anticuerpos Monoclonales/inmunología , Especificidad de Anticuerpos , Apoptosis/efectos de los fármacos , Factor Inductor de la Apoptosis/genética , Caspasa 14/inmunología , Caspasa 14/metabolismo , Línea Celular Tumoral , Cisplatino/farmacología , Estudios de Cohortes , Femenino , Células HeLa , Humanos , Inmunohistoquímica , Neoplasias Pulmonares/enzimología , Neoplasias Pulmonares/patología , Masculino , Ratones , Ratones Endogámicos BALB C , Modelos Moleculares , Datos de Secuencia Molecular , Recurrencia Local de Neoplasia/enzimología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tasa de Supervivencia , Transfección

RESUMEN

Keratinization is a kind of cell death called terminal differentiation and includes various patterns such as epidermal keratinization (EK), trichilemmal keratinization (TK), and shadow cell differentiation (SCD), whereas these have not been comparatively investigated from a standpoint of cell death. In the present study, surgically extirpated specimens of epidermal cyst, trichilemmal cyst, and pilomatricoma (10 cases in each) were subjected to immunohistochemistry for single-strand DNA (ssDNA), gamma-H2AX, cleaved caspase-3, cleaved lamin A, caspase-14, and CD138 to compare the modes of cell death and keratinization pattern. Transitional cells in pilomatricoma were immunoreactive, although not in whole part, for ssDNA and gamma-H2AX, and negative for cleaved caspase-3 and cleaved lamin A. Epidermal and trichilemmal cyst were negative for these 4 markers, except for ssDNA or cleaved lamin A in a small number of parakeratotic cells in a few cases. The keratinizing component showed caspase-14(+)/CD138(-) in epidermal cyst, caspase-14(-)/CD138(+) in trichilemmal cyst, and caspase-14(-)/CD138(-) in pilomatricoma. These results indicate that EK, TK, and SCD have a common property of apoptosis-like programmed cell death without caspase-3 activation or nuclear fragmentation. Meanwhile, they show different characteristics one another as follows: (A), DNA double-strand breaks occur in the transitional cells of SCD but not in EK/TK; and (B), EK, TK, and SCD can be distinguished by expression pattern of caspase-14 and CD138 in the keratinizing component.

Asunto(s)

Quiste Epidérmico/patología , Enfermedades del Cabello/patología , Queratinas/metabolismo , Pilomatrixoma/patología , Enfermedades de la Piel/patología , Neoplasias Cutáneas/patología , Biomarcadores de Tumor/análisis , Caspasa 14/biosíntesis , Muerte Celular , Diferenciación Celular , Roturas del ADN de Doble Cadena , Quiste Epidérmico/genética , Quiste Epidérmico/metabolismo , Enfermedades del Cabello/genética , Enfermedades del Cabello/metabolismo , Humanos , Inmunohistoquímica , Pilomatrixoma/genética , Pilomatrixoma/metabolismo , Enfermedades de la Piel/genética , Enfermedades de la Piel/metabolismo , Neoplasias Cutáneas/genética , Neoplasias Cutáneas/metabolismo , Sindecano-1/biosíntesis

RESUMEN

Exposure to airborne dust particles originated from seasonal Asian dust storms in Chinese and Mongolian deserts results in increased incidence of a range of diseases including asthma, contact dermatitis and conjunctivitis. The areas affected by Asian dust particles extend from East China to the west coast of North America. In order to study toxicological mechanisms in human skin, we evaluated the effects of dust particles collected during Asian dust storms (Asian dust particles) on gene expression in human epidermal keratinocytes (HEK). In HEK, exposure to Asian dust particles significantly increased gene expressions of cytochrome P450 1A1 (CYP1A1), CYP1A2, and CYP1B1, which is an indication of aryl hydrocarbon receptor (AHR) activation. In addition, Asian dust particles increased gene transcription of the cytokines IL-6, IL-8, and GM-CSF, which have broad pro-inflammatory and immunomodulatory properties. Asian dust particles significantly up-regulated expression of caspase 14 in HEK, suggesting that Asian dust particles directly affect keratinocyte differentiation. We also demonstrated that protein extract of pollen, a material frequently adsorbed onto Asian dust particles, potentially contributes to the increased transcription of IL-6, CYP1A1, CYP1A2, and CYP1B1. Taken together, these studies suggest that Asian dust particles can exert toxicological effects on human skin through the activation of the cellular detoxification system, the production of pro-inflammatory and immunomodulatory cytokines, and changes in the expression of proteins essential in normal epidermal differentiation.

Asunto(s)

Polvo , Queratinocitos/efectos de los fármacos , Material Particulado/farmacología , Transcripción Genética/efectos de los fármacos , Asia Occidental , Caspasa 14/biosíntesis , Caspasa 14/genética , Células Cultivadas , Sistema Enzimático del Citocromo P-450/biosíntesis , Sistema Enzimático del Citocromo P-450/genética , Perfilación de la Expresión Génica , Factor Estimulante de Colonias de Granulocitos y Macrófagos/biosíntesis , Factor Estimulante de Colonias de Granulocitos y Macrófagos/genética , Humanos , Inflamación/inducido químicamente , Interleucina-6/biosíntesis , Interleucina-6/genética , Interleucina-8/biosíntesis , Interleucina-8/genética , Queratinocitos/metabolismo , ARN Mensajero/biosíntesis , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa

RESUMEN

BACKGROUND: Current therapeutic approaches to salivary gland cancer are often associated with severe disfigurement and loss of glandular function, which are traumatic to the patients. Exploration of novel treatment approaches, such as gene therapy, is needed. MATERIALS AND METHODS: The human salivary gland cancer cell line HSG was transiently transfected with full length human caspase-14 cDNA. Photomicroscopy, BrdU assay, cell counting, MTT assay, and TUNEL assay were applied. To determine the tumorigenicity, tumor volume, tumor pathology and vascularization were analyzed in vivo. RESULTS: Cell growth and viability were inhibited significantly by transient caspase-14 expression. Caspase-14 expression resulted in a significant reduction of tumorigenicity. Importantly, a significant decrease in tumor blood vessel formation was observed. CONCLUSION: Salivary gland cancer cells underwent growth inhibition, cell death, and reduced tumorigenicity in vivo when exogenous caspase-14 was expressed, which could be due, in part, to an inhibitory effect of caspase-14 on tumor vascularization.

Asunto(s)

Adenocarcinoma/irrigación sanguínea , Adenocarcinoma/terapia , Caspasa 14/genética , Terapia Genética/métodos , Neoplasias de las Glándulas Salivales/irrigación sanguínea , Neoplasias de las Glándulas Salivales/terapia , Adenocarcinoma/enzimología , Adenocarcinoma/genética , Animales , Caspasa 14/biosíntesis , Caspasa 14/metabolismo , Procesos de Crecimiento Celular/genética , Línea Celular Tumoral , ADN de Neoplasias/biosíntesis , ADN de Neoplasias/genética , Femenino , Vectores Genéticos/genética , Humanos , Ratones , Ratones Desnudos , Neovascularización Patológica/enzimología , Neovascularización Patológica/genética , Neovascularización Patológica/patología , Plásmidos/genética , Neoplasias de las Glándulas Salivales/enzimología , Neoplasias de las Glándulas Salivales/genética , Transfección , Ensayos Antitumor por Modelo de Xenoinjerto

RESUMEN

Stratum corneum comprises corneocytes, derived from outer stratum granulosum during terminal differentiation, embedded in a lipid-enriched extracellular matrix, secreted from epidermal lamellar bodies. Permeability barrier insults stimulate rapid secretion of preformed lamellar bodies from the outer stratum granulosum, regulated through modulations in ionic gradients and serine protease (SP)/protease-activated receptor type 2 (PAR2) signaling. Because corneocytes are also required for barrier function, we hypothesized that corneocyte formation could also be regulated by barrier function. Barrier abrogation by two unrelated methods initiated a wave of cornification, assessed as TdT-mediated dUTP nick end-labeling-positive cells in stratum granulosum and newly cornified cells by electron microscopy. Because cornification was blocked by occlusion, corneocytes formed specifically in response to barrier, rather than injury or cell replacement, requirements. SP inhibitors and hyperacidification (which decreases SP activity) blocked cornification after barrier disruption. Similarly, cornification was delayed in PAR2(-/-) mice. Although classical markers of apoptosis [poly(ADP-ribose)polymerase and caspase (Casp)-3] remained unchanged, barrier disruption activated Casp-14. Moreover, the pan-Casp inhibitor Z-VAD-FMK delayed cornification, and corneocytes were structurally aberrant in Casp14(-/-) mice. Thus, permeability barrier requirements coordinately drive both the generation of the stratum corneum lipid-enriched extracellular matrix and the transformation of granular cells into corneocytes, in an SP- and Casp-14-dependent manner, signaled by PAR2.

Asunto(s)

Caspasa 14/biosíntesis , Caspasa 14/metabolismo , Epidermis/metabolismo , Regulación Enzimológica de la Expresión Génica , Receptor PAR-2/metabolismo , Animales , Apoptosis , Diferenciación Celular , Femenino , Concentración de Iones de Hidrógeno , Etiquetado Corte-Fin in Situ , Masculino , Ratones , Modelos Biológicos , Permeabilidad , Fenómenos Fisiológicos de la Piel

RESUMEN

The human placenta is responsible for the exchange of nutrients, gas and wastes through the trophoblast maternal-fetal barrier, which is formed by the fusion of villous cytotrophoblasts to form the continuous multinucleated syncytiotrophoblast separating the maternal and fetal circulations. Caspase-14 is a seemingly non-apoptotic caspase involved in keratinocyte differentiation and cornification. It is proposed that caspase-14 has a conserved role in cellular differentiation and a role in differentiation and fusion in the trophoblast. The human choriocarcinoma BeWo cell line was treated with staurosporine and forskolin to induce apoptosis and differentiation respectively. Staurosporine initiated apoptosis within 3 h of treatment, while apoptosis was completed following 6 h treatment. Caspase-14 gene and protein expression was unchanged throughout this process. During BeWo differentiation, caspase-14 mRNA was elevated after 48 h forskolin treatment, while its protein was increased after 24 h. Therefore, caspase-14 is up-regulated during trophoblast differentiation, as represented by the BeWo cell line. Moreover, caspase-14 may interact with other signalling molecules to facilitate differentiation. This new data confirms the potential for the BeWo cell line in the functional dissection of this unusual caspase and its prospective role in trophoblast differentiation.

Asunto(s)

Caspasa 14/biosíntesis , Trofoblastos/citología , Apoptosis , Diferenciación Celular , Línea Celular Tumoral , Colforsina/farmacología , Humanos , Queratinocitos/citología , Placenta/metabolismo , ARN Mensajero/metabolismo , Transducción de Señal , Estaurosporina/farmacología , Factores de Tiempo , Regulación hacia Arriba