RESUMEN
INTRODUCTION: Both oestrogen and progestin and the route of administration have been implicated in cardiovascular and thromboembolic risk in post menopausal hormone users. Transdermal preparations have been reported as safer indicating that liver derived metabolites of oestrogen may be important. The aim of our study was to investigate the in vitro effects of 17ß-estradiol, its metabolites, and norethisterone acetate (NETA) on the expression of coagulation genes in cultured human cells. METHODS: Human hepatocytes and human umbilical vein endothelial cells(HUVECS) were treated with 17ß-estradiol, estrone, 2-hydroxyestradiol (2-OH), NETA and NETA/17ß-estradiol (10nM) for 24hours. Fibrinogen, factor VII, prothrombin and plasminogen activator inhibitor -1 (PAI-1) mRNA expression was determined in hepatocyte cultures using TaqMan PCR. Tissue factor (TF), tissue factor pathway inhibitor (TFPI), tissue plasminogen activator (tPA) and PAI-1 expression was determined in HUVECS. Expression of estrogen receptors was also determined. RESULTS: Fibrinogen and factor VII mRNA expression was upregulated 2-4 fold by estradiol and estrone. Addition of NETA downregulated fibrinogen and prothrombin. PAI-1 expression in hepatocytes was upregulated by estrone, 2-OH, NETA and NETA/17ß-estradiol. In HUVECS, TF, TFPI and PAI-1 expression was upregulated by estrone but not by 17ß-estradiol. NETA upregulated TF, TFPI and tPA expression. Estrogen receptor status was unaffected by the addition of NETA. CONCLUSIONS: This data suggests a role for progestins in modifying the effects of oestrogen and its metabolites on coagulation gene expression which may contribute to the reduced thrombotic risk associated with transdermal preparations.
Asunto(s)
Coagulación Sanguínea/efectos de los fármacos , Anticonceptivos Sintéticos Orales/farmacología , Hepatocitos/efectos de los fármacos , Células Endoteliales de la Vena Umbilical Humana/efectos de los fármacos , Noretindrona/análogos & derivados , Coagulación Sanguínea/genética , Células Cultivadas , Estradiol/análogos & derivados , Estradiol/farmacología , Estrona/farmacología , Factor VII/genética , Factor VII/metabolismo , Fibrinógeno/genética , Fibrinógeno/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Hepatocitos/metabolismo , Células Endoteliales de la Vena Umbilical Humana/metabolismo , Humanos , Lipoproteínas/genética , Lipoproteínas/metabolismo , Noretindrona/farmacología , Acetato de Noretindrona , Inhibidor 1 de Activador Plasminogénico/genética , Inhibidor 1 de Activador Plasminogénico/metabolismo , Protrombina/genética , Protrombina/metabolismo , ARN Mensajero/metabolismo , Reacción en Cadena en Tiempo Real de la Polimerasa , Receptores de Estrógenos/efectos de los fármacos , Receptores de Estrógenos/genética , Receptores de Estrógenos/metabolismo , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Tromboplastina/genética , Tromboplastina/metabolismo , Factores de Tiempo , Activador de Tejido Plasminógeno/genética , Activador de Tejido Plasminógeno/metabolismoRESUMEN
MicroRNAs are a group of small non-coding RNAs approximately 22 nucleotides in length. Recent work has shown differential expression of mature microRNAs in human cancers. We characterized the alteration in expression of a select group of microRNAs in primary peritoneal carcinoma relative to matched cases of ovarian serous carcinoma. MicroRNA expression was analysed using semi-quantitative stem-loop RT-PCR on a set of 34 formalin-fixed paraffin-embedded samples. Protein expression of p53 and bcl-2 was quantified in the corresponding tissue microarray. We provide definitive evidence that there is downregulation of a select group of microRNAs in tumours meeting Gynaecological Oncology Group criteria for primary peritoneal carcinoma relative to ovarian serous carcinoma. Specifically, we show decreased p53 expression and downregulation of miR-195 and miR-497 from the microRNA cluster site at chromosome 17p13.1 in primary peritoneal carcinoma relative to ovarian serous carcinoma. miR-195 and miR-497 may have potential roles as tumour-suppressor genes in primary peritoneal tumourigenesis.
Asunto(s)
Carcinoma/genética , Cromosomas Humanos Par 17 , Regulación Neoplásica de la Expresión Génica , MicroARNs/análisis , Neoplasias Ováricas/genética , Neoplasias Peritoneales/genética , Adulto , Anciano , Anciano de 80 o más Años , Carcinoma/química , Carcinoma/patología , Regulación hacia Abajo , Femenino , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Neoplasias Ováricas/química , Neoplasias Ováricas/patología , Neoplasias Peritoneales/química , Neoplasias Peritoneales/patología , Proteínas Proto-Oncogénicas c-bcl-2/análisis , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Matrices Tisulares , Proteína p53 Supresora de Tumor/análisisRESUMEN
MicroRNAs are a group of small non-coding RNAs approximately 22 nucleotides in length. Recent work has shown differential expression of mature microRNAs in human cancers. Production and function of microRNAs require coordinated processing by proteins of the microRNA machinery. Dicer and Drosha (RNase III endonucleases) are essential components of the microRNA machinery. Recently, the ribosome anti-association factor eIF6 has also been found to have a role in microRNA-mediated post-transcriptional silencing. We characterized the alterations in the expression of genes encoding proteins of microRNA machinery in ovarian serous carcinoma. Protein expression of eIF6 and Dicer was quantified in a tissue microarray of 66 ovarian serous carcinomas. Dicer, Drosha and eIF6 mRNA expression was analysed using quantitative reverse transcription-PCR on an independent set of 50 formalin-fixed, paraffin-embedded ovarian serous carcinoma samples. Expression profiles of eIF6 and Dicer were correlated with clinicopathological and patient survival data. We provide definitive evidence that eIF6 and Dicer are both upregulated in a significant proportion of ovarian serous carcinomas and are associated with specific clinicopathological features, most notably low eIF6 expression being associated with reduced disease-free survival. The status of eIF6 and proteins of the microRNA machinery may help predict toxicity and susceptibility to future interfering RNA-based therapy.
Asunto(s)
Cistadenocarcinoma Seroso/patología , Factores Eucarióticos de Iniciación/biosíntesis , Neoplasias Ováricas/patología , Ribonucleasa III/biosíntesis , Cistadenocarcinoma Seroso/metabolismo , Cistadenocarcinoma Seroso/mortalidad , Femenino , Expresión Génica , Perfilación de la Expresión Génica , Humanos , Inmunohistoquímica , Estimación de Kaplan-Meier , Persona de Mediana Edad , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/mortalidad , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Matrices TisularesAsunto(s)
Recurrencia Local de Neoplasia/genética , Neoplasias Ováricas/genética , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Persona de Mediana Edad , Recurrencia Local de Neoplasia/metabolismo , Recurrencia Local de Neoplasia/patología , Estadificación de Neoplasias , Análisis de Secuencia por Matrices de Oligonucleótidos , Neoplasias Ováricas/metabolismo , Neoplasias Ováricas/patología , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Hormone therapy (HT) increases the risk of cardiovascular and thromboembolic disease in post-menopausal women. Recent studies have suggested that prothrombotic mechanisms are likely to be involved. Transdermal HT avoids the first-pass effect of oestrogen in the liver and may have a less marked effect on the haemostatic system than equivalent oral preparations. The majority of studies have compared HT preparations that have different formulations as well as routes of administration. We investigated changes in the haemostatic system in post-menopausal women using two pharmacologically similar HT preparations, which differed only in their route of administration. Three hundred forty-four healthy post-menopausal women were randomised to six months treatment with either a transdermal matrix patch containing 25 microg 17beta-estradiol/125 microg norethisterone acetate (NETA) applied every 3-4 days, or an equivalent oral preparation (estradiol 1 mg and NETA 0.5 mg given once daily). Oral treatment significantly reduced fibrinogen (p < 0.003), factor VIIc (FVIIc) (p < 0.00001), and antithrombin (AT) levels (p < 0.005); the effects in the transdermal group were less marked with no reduction in fibrinogen levels and lesser effect on FVIIc (p < 0.03) compared with oral treatment. Treatment type significantly affected fibrinolysis with lower plasmin-anti-plasmin (PAP) levels in the transdermal group (p < 0.003) and lower plasminogen activator inhibitor-1 antigen (PAI-1) (p < 0.012) and tissue plasminogen activator (tPA) antigen levels in the oral group (p < 0.002). Prothrombin fragment 1.2 and activated protein C (APC) resistance were not affected by either treatment. Transdermal HT has a less marked effect on coagulation than an equivalent oral preparation. Randomised trials are required to investigate whether this translates into less risk of cardiovascular and thromboembolic disease.
Asunto(s)
Coagulación Sanguínea/efectos de los fármacos , Estradiol/administración & dosificación , Terapia de Reemplazo de Estrógeno , Noretindrona/análogos & derivados , Posmenopausia , Administración Cutánea , Administración Oral , Anciano , Antígenos/sangre , Antitrombinas/metabolismo , Enfermedades Cardiovasculares/sangre , Enfermedades Cardiovasculares/inducido químicamente , Combinación de Medicamentos , Estradiol/efectos adversos , Terapia de Reemplazo de Estrógeno/efectos adversos , Factor VII , Femenino , Fibrinógeno/metabolismo , Fibrinolisina/metabolismo , Fibrinólisis/efectos de los fármacos , Humanos , Irlanda , Persona de Mediana Edad , Noretindrona/administración & dosificación , Noretindrona/efectos adversos , Acetato de Noretindrona , Fragmentos de Péptidos/sangre , Protrombina , Factores de Riesgo , Factores de Tiempo , Activador de Tejido Plasminógeno/sangre , Resultado del Tratamiento , alfa 2-Antiplasmina/metabolismoRESUMEN
The aim of this study was to identify genomic aberrations in endometrial cancer cells treated with the phyto-estrogenic compounds tectorigenin, irigenin and apigenin and to compare with those treated with beta-estradiol using array-based comparative genomic hybridisation (array CGH). The microarray contains 287 targets and includes telomeres, microdeletions, oncogenes and tumour suppressor genes and has increased mapping resolution compared to conventional CGH. An endometrial cancer cell line (Ishikawa) was cultured and treated with the phyto-estrogens. Treated cells were examined using the CGH microarray. Over 20 % of the array genes were aberrated in the cells treated with beta-estradiol, tectorigenin and irigenin compared to 3 % in those treated with the same concentration of apigenin. Protein kinase c zeta form, insulin, insulin receptor and protein-tyrosine phosphatase non-receptor-type 1 which are involved in insulin metabolism were aberrated by tectorigenin and irigenin. Apigenin may play a role in the treatment of endometrial cancer and in the treatment of postmenopausal women. Further studies in normal endometrium and primary endometrial cancer cells are needed to elucidate the role of the phyto-estrogens.
Asunto(s)
Aberraciones Cromosómicas , Neoplasias Endometriales/genética , Iridaceae , Fitoestrógenos/farmacología , Fitoterapia , Apigenina/farmacología , Línea Celular Tumoral/efectos de los fármacos , ADN de Neoplasias/análisis , Estradiol/farmacología , Femenino , Perfilación de la Expresión Génica , Humanos , Isoflavonas/farmacología , Extractos Vegetales/farmacología , ARN Mensajero/análisisRESUMEN
OBJECTIVES: The aim of this study was to examine the expression of oestrogen regulated genes in premenopausal and postmenopausal normal and malignant endometrial specimens. The molecular mechanisms and the role of these genes in endometrial carcinogenesis are poorly understood. METHODS: Normal and malignant endometrial specimens were collected from patients undergoing hysterectomy. Real time TaqMan PCR was used to examine the mRNA expression levels of oestrogen receptor a (ERa) and b (ERb), progesterone receptor (PR), insulin like growth factor 1 (IGF-1) and vascular endothelial growth factor (VEGF). RESULTS: Expression analysis was carried out on 60 patients. ERa was more predominantly expressed in the endometrial samples than ERb, 28% of the specimens did not express ER. Normal pre and postmenopausal tissue expressed higher levels of ERa, PR and IGF-1 than malignant tissue. ERa and PR expression was significantly higher in the proliferative phase endometrium compared to the secretory phase (P < 0.05). PR mRNA expression was significantly correlated with ERa in all tissue types. CONCLUSIONS: ERa expression may play an important role in the regulation of PR in normal and malignant endometrium. Further work is needed to establish if IGF-1 plays a role in a subset of endometrial cancers and if isoforms of VEGF play a role in endometrial cancer.
Asunto(s)
Adenocarcinoma/metabolismo , Neoplasias Endometriales/metabolismo , Endometrio/metabolismo , Regulación Neoplásica de la Expresión Génica , Receptores de Estrógenos/genética , Adenocarcinoma/patología , Adulto , Anciano , Anciano de 80 o más Años , Neoplasias Endometriales/patología , Endometrio/patología , Femenino , Humanos , Factor I del Crecimiento Similar a la Insulina/genética , Factor I del Crecimiento Similar a la Insulina/metabolismo , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Posmenopausia , Premenopausia , ARN Mensajero/análisis , Receptores de Estrógenos/sangre , Receptores de Estrógenos/metabolismo , Receptores de Progesterona/sangre , Receptores de Progesterona/genética , Receptores de Progesterona/metabolismo , Factor A de Crecimiento Endotelial Vascular/sangre , Factor A de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
The aim of this study was to use the (3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt) (MTS) assay to determine the response of 17 endometrial and 27 cervical tumours to cytotoxic drugs. Tumour samples were taken at surgery and cultured using the explant technique. Cells were incubated with chemotherapy drugs. The MTS cytotoxicity assay was carried out to ascertain the response to the drugs and correlated retrospectively to the clinical outcome. Tumours of similar stage and grade displayed heterogeneity in their responses to the drugs. A total of 88 of 90 tumours (97.8%), including data from an earlier study of 44 ovarian tumour samples yielded chemosensitivity data. A total of 45 specimens were evaluable for in vitro-in vivo correlations. In vitro sensitivity was associated with clinical response in 26 of 30 patients and in vitro resistance with progressive disease or death in 14 of 15 patients. A randomised prospective trial should be carried out to validate chemosensitivity/resistance testing.