RESUMEN
Cancer chemotherapy can fail in many ways. One of the most significant is the development of multiple drug resistance (MDR), which constitutes a serious clinical problem. The development of MDR relates to the expression of a major membrane pump, P-glycoprotein (P-gp). Thus, currently one of the goals of experimental and clinical oncology is to decrease its activity. So far, many different P-gp inhibitors are available, but their efficacy is still questionable and requires further study. The aim of our study was to assess an impact of classical P-gp inhibitors (verapamil and cyclosporin A) in the reversion of multidrug resistance in canine mammary cancer cells. We used two cell lines isolated from mammary tumors and two cell lines isolated from their lung metastases. All of them showed P-gp over-expression confirmed using Real-time rt-PCR, Skan(R) screening station and confocal microscopy. The FACS analysis showed that in three of the examined cell lines, treatment with verpamil/cyclosporin A was ineffective to reverse cancer chemoresistance. However, more studies in this field are required.
Asunto(s)
Adenocarcinoma/tratamiento farmacológico , Ciclosporina/farmacología , Neoplasias Mamarias Animales/tratamiento farmacológico , Verapamilo/farmacología , Vinblastina/farmacología , Subfamilia B de Transportador de Casetes de Unión a ATP/antagonistas & inhibidores , Animales , Antineoplásicos , Línea Celular Tumoral , Enfermedades de los Perros/tratamiento farmacológico , Perros , Sinergismo Farmacológico , Femenino , Regulación Neoplásica de la Expresión Génica/fisiología , Vinblastina/administración & dosificaciónRESUMEN
Because canine mammary tumours constitute a serious clinical problem and there are no good prognostic markers (only histopathological variables are used), the aim of the presented study was to find new malignancy markers as well as to identify intracellular pathways and biological processes characteristic for canine mammary malignancy. We compared gene expression of the most malignant mammary tumours (poorly differentiated cancers of the 3rd grade of malignancy) with less malignant tumours (well differentiated cancers of the 1st grade of malignancy). The results of our study indicated that in dogs the number of tumour-infiltrating myeloid cells or expression of myeloid-specific antigens by cancer cells is related to the cancer progression and may constitute a new marker of malignancy, however further studies in this field are required.
Asunto(s)
Biomarcadores de Tumor/genética , Enfermedades de los Perros/genética , Inflamación/veterinaria , Neoplasias Mamarias Animales/genética , Animales , Biomarcadores de Tumor/biosíntesis , Enfermedades de los Perros/diagnóstico , Enfermedades de los Perros/metabolismo , Enfermedades de los Perros/patología , Perros , Femenino , Regulación Neoplásica de la Expresión Génica , Inmunohistoquímica/veterinaria , Inflamación/diagnóstico , Inflamación/genética , Inflamación/patología , Neoplasias Mamarias Animales/diagnóstico , Neoplasias Mamarias Animales/metabolismo , Neoplasias Mamarias Animales/patología , Clasificación del Tumor , Análisis de Secuencia por Matrices de Oligonucleótidos/veterinaria , ARN Neoplásico/química , ARN Neoplásico/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinariaRESUMEN
Feline vaccine associated fibrosarcomas are the second most common skin tumor in cats. Methods of treatment are: surgery, chemotherapy and radiotherapy. Nevertheless, the usage of cytostatics in feline vaccine associated sarcoma therapy is limited due to their adverse side effects, high toxicity and low biodistribution after i.v. injection. Therefore, much research on new therapeutic drugs is being conducted. In human medicine, the chick embryo chorioallantoic membrane (CAM) model is used as a cheap and easy to perform assay to assess new drug effectiveness in cancer treatment. Various human cell lines have different tumors growth on CAM. In veterinary medicine such model has not been described yet. In the present article derivation of feline vaccine associated fibrosarcoma cell line and its growth on CAM is described. The cell line and the tumor grown were confirmed by histopathological and immunohistochemical examination. As far as we believe, this is the first attempt to create such model, which may be used for further in vivo studies in veterinary oncology.
Asunto(s)
Enfermedades de los Gatos/patología , Membrana Corioalantoides/metabolismo , Modelos Animales de Enfermedad , Fibrosarcoma/veterinaria , Neoplasias Cutáneas/veterinaria , Células Tumorales Cultivadas , Vacunación/efectos adversos , Animales , Gatos , Embrión de Pollo , Pollos , Membrana Corioalantoides/patología , Fibrosarcoma/metabolismo , Fibrosarcoma/patología , Neoplasias Cutáneas/metabolismo , Neoplasias Cutáneas/patología , Vacunación/veterinariaRESUMEN
Ghrelin is a 28 amino-acid multi-functional peptide hormone, which was identified as a natural ligand of the growth hormone secretagogue receptor (GHS-R). Pituitary growth hormone-releasing activity in both animals and humans has been well documented. It has various biological functions, including regulation of appetite and body weight, control of energy homeostasis, modulation of cardiovascular and gastrointestinal system and anti-inflammatory effect. However, both ghrelin and its receptor (GHS-R) are widely distributed in various tumors, which strongly implies their role in neoplastic cell growth trough autocrine/paracrine mechanism. Multiple studies have demonstrated the role of ghrelin in cancer cells proliferation, differentiation, invasiveness and apoptosis inhibition. The ghrelin axis is more complex than it was originally thought and consist of several compounds that might interact with each other and affect ghrelin activities. Here, we provide an overview of the ghrelin and its receptor role in tumor progression.
Asunto(s)
Regulación Neoplásica de la Expresión Génica/fisiología , Ghrelina/metabolismo , Neoplasias/metabolismo , Receptores de Ghrelina/metabolismo , Animales , Ghrelina/genética , Receptores de Ghrelina/genéticaRESUMEN
Canine mammary gland has been identified as a major site of the extrapituitary growth hormone (GH) production. This finding is linked to its role in tumourigenesis of the mammary gland. Our previous studies indicated the role of GH and GH receptor (GHR) in regulation of proliferation and apoptosis. Thus, we have optimized the ghr RNA interference method in canine mammary carcinoma cell line CMT-U27. We have analysed the effect of GHR reduction on the intracellular signalling and the cell cycle and apoptosis. The results showed that GHR reduction decreased the p-ERK1/2 expression and caused increase of apoptosis and decrease in number of cells at S and G2M phases. This study indicates that GHR besides proliferative effect promotes growth by increasing cell survival. It can tilt the balance between proliferation and death in cancer cells.
Asunto(s)
Apoptosis , Enfermedades de los Perros/metabolismo , Neoplasias Mamarias Animales/metabolismo , Neoplasias Mamarias Experimentales/metabolismo , Receptores de Somatotropina/metabolismo , Análisis de Varianza , Animales , Línea Celular Tumoral , Bases de Datos de Ácidos Nucleicos , Enfermedades de los Perros/patología , Perros , Femenino , Inmunohistoquímica/veterinaria , Neoplasias Mamarias Animales/patología , Interferencia de ARN , Receptores de Somatotropina/genéticaRESUMEN
Several years ago, the presence of macrophages in the tumor microenvironment was thought to be an inflammatory response to kill the cancer cells. Now, this is clear that the inflammatory cells that exit blood vessels and migrate to the tumor tissue play an important role in cancer progression. Various cells present in the tumor microenvironment enhance cancer growth and invasiveness by secretion of tumor-enhancing products. That is why tumors should not be treated as only aggregates of cancer cells but as separate structures. Macrophages form a major component of the inflammatory infiltration in tumors, where they are termed tumor-associated macrophages (TAMs). To the best of our knowledge, up-to-date there were no studies on tumor associated macrophages and the role of the tumor microenvironment in tumor invasion/metastasis in dogs. This is the first study performed to asses if the number of TAMs and expression of MCSF-R (macrophages colony stimulating factor receptor) and CD14 (LPS co-receptor) are associated with the grade of tumor malignancy and its ability to metastasize. We have performed immunohistochemical analysis of 50 canine mammary adenocarcinomas of various grade of malignancy (1st, 2nd, 3rd) and tumors that gave local or distant metastases. The results indicate that in dogs, similarly to humans and mice, the number of tumor associated macrophages is related to the cancer ability to metastasize. Our results also indicate that the expression of MCSF-R and, what is particularly new finding, CD14 is associated with tumor malignancy and its ability to metastasize. Hence, these molecules play a role in tumor progression, metastasis and microenvironment interactions. These results show that in dogs we should treat the tumor as a whole organ rather than just try to eliminate the cancer cells.
Asunto(s)
Adenocarcinoma/veterinaria , Enfermedades de los Perros/patología , Regulación Neoplásica de la Expresión Génica/fisiología , Macrófagos/metabolismo , Neoplasias Mamarias Animales/metabolismo , Receptor de Factor Estimulante de Colonias de Macrófagos/metabolismo , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Animales , Perros , Femenino , Receptores de Lipopolisacáridos/metabolismo , Macrófagos/citologíaRESUMEN
Canine mammary sarcomas are usually very aggressive and easily metastasize. Unfortunately the biology of this type of tumor is not well known because they are a very rare type of tumors. The aim of this study was to find differences in gene expression patterns in canine mammary osteosarcomas (malignant) versus osteomas (benign) using DNA microarrays. Our microarray experiment showed that 11 genes were up-regulated in osteosarcoma in comparison to osteoma whereas 36 genes were down-regulated. Among the up-regulated genes were: PDK1, EXT1, and EIF4H which are involved in AKT/PI3K and GLI/Hedgehog pathways. These genes play an important role in cell biology (cancer cell proliferation) and may be essential in osteosarcoma formation and development. Analyzing the down-regulated genes, the most interesting seemed to be HSPB8 and SEPP1. HSPB8 is a small heat shock protein that plays an important role in cell cycle regulation, apoptosis, and breast carcinogenesis. Also SEPP1 may play a role in carcinogenesis, as its down-regulation may induce oxidative stress possibly resulting in carcinogenesis. The preliminary results of the present study indicate that the up-regulation of three genes EXT1, EIF4H, and PDK1 may play an essential role in osteosarcoma formation, development and proliferation. In our opinion the cross-talk between GLI/Hedgehog and PI3K/AKT pathways may be a key factor to increase tumor proliferation and malignancy.
Asunto(s)
Enfermedades de los Perros/metabolismo , Regulación Neoplásica de la Expresión Génica/fisiología , Neoplasias Mamarias Animales/metabolismo , Osteosarcoma/metabolismo , Animales , Perros , Regulación hacia Abajo , Femenino , Perfilación de la Expresión Génica , Neoplasias Mamarias Animales/genética , Osteosarcoma/genética , Reacción en Cadena de la Polimerasa , ARN/genética , ARN/metabolismo , Regulación hacia ArribaRESUMEN
There are many reasons that lead to failure of cancer chemotherapy. Cancer has the ability to become resistant to many different types of drugs. Increased efflux of drug, enhanced repair/increased tolerance to DNA damage, high antiapoptotic potential, decreased permeability and enzymatic deactivation allow cancer cell survive the chemotherapy. Treatment can lead to the death of most tumor cells (drug-sensitive), but some of them (drug-resistant) survive and grow again. These tumor cells may arise from stem cells. There are many studies describing human experiments with multidrug resistance, especially in breast cancer. Unfortunately, studies of canine or feline ABC super family members are not as extensive as in human or mice and they are limited to several papers describing PGP in mammary cancer, cutaneous mast cell tumors and lymphoma. Multidrug resistance is one of the most significant problems in oncology today. The involvement of many different, not fully recognized, mechanisms in multidrug resistance of cancer cells makes the development of effective methods of therapy very difficult. Understanding the mechanisms of drug resistance in cancer cells may improve the results of treatment. This review article provides a synopsis of all aspects that refer to cancer cell resistance to antitumor drugs.
Asunto(s)
Antineoplásicos/uso terapéutico , Neoplasias/veterinaria , Insuficiencia del Tratamiento , Animales , Resistencia a Antineoplásicos , Humanos , Neoplasias/tratamiento farmacológicoRESUMEN
In light of the high incidence of mammary cancer in dogs and completion of the canine genome sequencing, the new possibilities of gene profiling by using DNA microarrays give hope to veterinary oncology. The cell lines isolated from mammary tumors are a valuable tool in developing and testing new pathway-specific cancer therapeutics. Differential cytometric analysis of 6 canine mammary cancer cell lines was performed. We divided cell lines into 3 groups based on their phenotype: 2 lines with high proliferative potential, 2 lines with high antiapoptotic potential, and 2 lines with high metastatic potential. DNA microarray analysis revealed common genes for cell lines of each group. We found that genes encoding the receptors for growth hormone and ghrelin are related to high proliferation rate, while ABR (active BCR-related) and TMD1 (TM2 domain containing 1) genes are related to a high antiapoptotic potential of the cancer cells. Metastatic properties of mammary cancer cells seem to be associated with elevated expression of PGP (P glycoprotein), SEMA3B (semaphorin 3B), and STIM1 (stromal interaction molecule 1).
Asunto(s)
Enfermedades de los Perros/genética , Perfilación de la Expresión Génica , Neoplasias Mamarias Animales/genética , Adenocarcinoma/genética , Adenocarcinoma/patología , Adenocarcinoma/veterinaria , Animales , División Celular , Línea Celular Tumoral , ADN de Neoplasias/genética , Perros , Femenino , Cinética , Neoplasias Mamarias Animales/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , Fenotipo , PloidiasRESUMEN
Metastasis is a final step in the progression of mammary gland cancer, usually leading to death. Potentially, a molecular signature of metastasis can be defined via comparison of primary tumors with their metastases. Currently, there is no data in the literature regarding the molecular portrait of metastases in dogs and only few reports regarding human cancer. This is the first report describing the transcriptomic signature of canine cancer metastatic cells. Two adenocarcinoma cell lines isolated from the canine mammary gland (CMT-W1 and CMT-W2) were compared with cell lines isolated from their lung metastases (CMT-W1M and CMT-W2M) with regards to the following cytometric parameters: cell cycle, ploidy, Bcl-2 expression, susceptibility to induced apoptosis, and transcriptomic profile. Cytometric analyses revealed significant differences in cell cycle and antiapoptotic potential between the examined cells. Using oligonucleotide microarrays, we found 104 up-regulated genes in the metastatic cell line CMT-W1M and 21 up-regulated genes in the primary CMT-W1 cell line. We also found 83 up-regulated genes in the CMT-W2M cell line and only 21 up-regulated genes in the CMT-W2 cell line. Among the up-regulated genes in both metastatic cell lines, we found 15 common genes. These differently expressed genes are involved mainly in signal transduction, cell structure and motility, nucleic acid metabolism, developmental processing, and apoptosis (GHSR, RASSF1, ARF1GAP, WDR74, SMOC2, SFRP4, DIAPH1, FSCN1, ALX4, SNX15, PLD2, WNT7B, POU6F2, NKG7, and POLR2F). Seven of them are involved in a cellular pathway dependent on ghrelin via growth hormone secretagogue receptor (GHSR). Our results suggest that this pathway may be essential for mammary cancer cells to have a metastatic potential.
Asunto(s)
Adenocarcinoma/genética , Biomarcadores de Tumor/genética , Perfilación de la Expresión Génica , Neoplasias Pulmonares/genética , Neoplasias Mamarias Animales/genética , Adenocarcinoma/metabolismo , Adenocarcinoma/patología , Animales , Apoptosis , Biomarcadores de Tumor/metabolismo , Western Blotting , Perros , Femenino , Técnica del Anticuerpo Fluorescente , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundario , Neoplasias Mamarias Animales/metabolismo , Neoplasias Mamarias Animales/patología , Análisis de Secuencia por Matrices de Oligonucleótidos , Ploidias , Pronóstico , ARN Mensajero/genética , ARN Mensajero/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Células Tumorales CultivadasRESUMEN
Tumor-derived cell lines are widely used as in vitro cancer models. Cell lines historically served as the primary experimental model systems for exploration of tumor cell biology and pharmacology. However, their ability to accurately reflect the phenotype and genotype of the parental histology remains questionable, given the prevalence of documented cell line-specific cytogenetic changes. Sometimes cell line studies are interpreted in the context of artifacts introduced by selection and establishment of cell lines in vitro. This complication has led to difficulties in the extrapolation of biology observed in cell lines to tumor biology in vivo. The aim of our study was to compare gene expression profiles in canine mammary tumor tissue and cell cultures derived from those tumors using cDNA microarrays. Tumors of two different origins were used; chondrosarcoma and adenocarcinoma and their primary cell cultures. It has been found that cell culture gene expression profiles closely resembled those of their corresponding in vivo tumor. In adenocarcinoma and chondrosarcoma only 6.0% and 2.7% of genes respectively, have shown significant difference in expression. In the most cases the difference concerned up-regulation of gene expression in cell lines, particularly genes involved in: protein metabolism and modification, signal transduction and nucleotide, nucleoside and nucleic acid metabolism. These experiments revealed that transcriptome of our primary cell culture corresponds to transcriptome of its parental tumor tissue and for this reason cell culture represents the reliable in vitro model for oncogenomic and pharmacogenomic studies.
Asunto(s)
Adenocarcinoma/metabolismo , Línea Celular Tumoral/metabolismo , Condrosarcoma Mesenquimal/metabolismo , Perfilación de la Expresión Génica , Neoplasias Mamarias Animales/metabolismo , Adenocarcinoma/genética , Animales , Condrosarcoma Mesenquimal/genética , Perros , Femenino , Regulación Neoplásica de la Expresión Génica , Neoplasias Mamarias Animales/genética , Análisis de Secuencia por Matrices de OligonucleótidosRESUMEN
The aim of the study was to identify the genes responsible for the high growth rate and antiapoptotic potential in selected canine mammary cancer cells. cDNA canine microarrays were used to compare the transcriptome in simple carcinoma CMT-U27 and spindle-cell tumor CMT-U309 cell lines. In CMT-U27 cell line the growth rate (shorter cell cycle), anti-apoptotic potential (higher expression of Bcl-2) was higher and spontaneous and induced apoptosis was lower. Comparison of transcriptomes revealed 333 genes which expression differed similarly. We focused on genes involved in cell proliferation, adhesion and apoptosis, and selected 29 of them. The high growth rate and anti-apoptotic potential in CMT-U27 cells was associated with enhanced expression of genes (at the level of transcripts) involved in Ca(2+) signaling pathway (Calmodulin 1, 2, 3 and SPSB2) and growth hormone cellular pathway. The low-proliferative and pro-apoptotic phenotype of CMTU309 cells was more dependent on TGFbeta, neuregulin 1 pathways and adhesion-related molecules.