RESUMEN

The primary aim of this study is to evaluate the clinical significance of E-cadherin protein expression and the methylation status in CDH1 promoter in endometrial cancer. The expression of E-cadherin and methylation in its promoter region was analyzed, retrospectively, in 152 clinical tissue samples from patients with endometrial lesions. We found that the hypermethylation of CDH1 promoter, which caused low expression of E-cadherin in endometrial cancer, was associated with not only clinicopathological progress of endometrial cancer but also with the overall 5-year clinical survival rate. The findings provide the potential therapeutic and prognostic target molecule for patients with endomethrial cancer.

Asunto(s)

Cadherinas/análisis , Cadherinas/genética , Carcinoma/química , Carcinoma/genética , Metilación de ADN , Neoplasias Endometriales/química , Neoplasias Endometriales/genética , Regiones Promotoras Genéticas , Antígenos CD , Carcinoma/mortalidad , Carcinoma/patología , China , Regulación hacia Abajo , Neoplasias Endometriales/mortalidad , Neoplasias Endometriales/patología , Femenino , Regulación Neoplásica de la Expresión Génica , Humanos , Estimación de Kaplan-Meier , Persona de Mediana Edad , Invasividad Neoplásica , Estadificación de Neoplasias , Pronóstico , Modelos de Riesgos Proporcionales , Estudios Retrospectivos , Medición de Riesgo , Factores de Riesgo , Factores de Tiempo

RESUMEN

OBJECTIVE: To study the therapeutic effect of recombinant adeno-associated virus carrying human endostatin gene therapy on endometriosis in mice model. METHODS: Recombinant adeno-associated virus vector carrying human endostatin gene and enhanced green fluorescent proteins gene (rAAV2-endostatin-EGFP) was constructed. Endometrium was from 12 patients with leiomyoma undergoing hysterectomy in Second Hospital, Tianjin Medical University between November and December 2008. Endometriosis models of nude mice were established by transplanting human endometrial fragments intooperitoneal surface. After 1 week, those 60 mice were divided into 3 groups: treatment group including 20 mice injected with rAAV2-endostatin-EGFP to ectopic lesion, control group including 20 mice injected with rAAV2-EGFP to ectopic lesion and blank control group including 20 mice injected with phosphate buffered saline (PBS) to the ectopic lesion. At 1, 2 and 3 weeks after treatment, those mice underwent laparotomy to observe the location and size of ectopic lesion in abdominal cavity. The expression of endostain protein, number of gland, microvessel density (MVD) and vascular endothelial growth factor (VEGF) were measured in ectopic lesions. The serum level of estradiol and progesterone were detected in nude mice among every groups. RESULTS: (1) All endometriosis of nude mice models were established successfully through peritoneum transplanting. After 1 week's treatment, flat lesion nodes, decreased gland number and narrow and atrophy glandular cavity were observed by light microscope. (2) The endostatin gene was transferred into nude mice successfully and expressed effectively. It was observed that endostatin protein expression was shown with enhanced green fluorescent proteins in ectopic lesion. (3) Glands number of ectopic lesion in rAAV2-endostatin-EGFP group (7.8 ± 1.9, 7.0 ± 1.5 and 5.5 ± 1.7) were significantly less than 10.1 ± 1.7, 10.2 ± 2.0 and 9.8 ± 2.4 in rAAV2-EGFP control group and 10.2 ± 2.2, 10.0 ± 2.0 and 9.7 ± 2.2 in PBS control group at 1, 2 and 3 weeks after treatment (all P < 0.05). Glands number of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P < 0.05). (4) MVD of ectopic lesion in rAAV2-endostatin-EGFP group (12.2 ± 1.5, 11.4 ± 2.1 and 9.0 ± 1.4) was significantly less than those at rAAV2-EGFP control group (16.5 ± 1.7, 16.5 ± 1.9 and 16.9 ± 1.9) and PBS control group (16.2 ± 1.6, 16.0 ± 1.6 and 16.3 ± 1.7) at 1, 2 and 3 weeks after treatment (all P < 0.05). MVD of ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P < 0.05). (5) The rate and density of VEGF expression at ectopic lesion in rAAV2-endostatin-EGFP group (35%, 30%, 25% and 1.60 ± 0.43, 1.33 ± 0.30, 1.03 ± 0.36) were significantly less than those at rAAV2-EGFP control group (80%, 75%, 85% and 2.43 ± 0.53, 2.43 ± 0.29, 2.66 ± 0.45) and PBS control group (85%, 90%, 90% and 2.36 ± 0.53, 2.64 ± 0.57, 2.53 ± 0.52) at one 1, 2 and 3 weeks after treatment (all P < 0.05). The expression of VEGF at ectopic lesion in rAAV2-endostatin-EGFP group at 3 weeks was significantly less than those at 1 and 2 weeks after treatment (P < 0.05). (6) The level of estradial and progesterone in serum of nude mice of rAAV2-endostatin-EGFP group [E(2): (48 ± 7) pmol/L, P: (61 ± 8) nmol/L] did not reach statistical difference when compared with those at rAAV2-EGFP control group [E(2): (50 ± 9) pmol/L, P: (60 ± 10) nmol/L] and PBS control group [E(2): (48 ± 7) pmol/L, P: (58 ± 10) nmol/L, P > 0.05]. CONCLUSIONS: The recombinant adeno-associated virus carrying human endostatin gene therapy could inhibit angiogenesis at endometriotic lesions and not influence steroid level. The antiangiogenic gene therapy might become a novel option for endometriosis.

Asunto(s)

Inhibidores de la Angiogénesis/uso terapéutico , Endometriosis/terapia , Endostatinas/genética , Terapia Genética/métodos , Inhibidores de la Angiogénesis/genética , Animales , Dependovirus/genética , Modelos Animales de Enfermedad , Endometriosis/genética , Endometriosis/metabolismo , Endostatinas/metabolismo , Endostatinas/uso terapéutico , Femenino , Vectores Genéticos , Proteínas Fluorescentes Verdes/genética , Proteínas Fluorescentes Verdes/metabolismo , Humanos , Ratones , Ratones Endogámicos BALB C , Ratones Desnudos , Neovascularización Patológica , Proteínas Recombinantes/uso terapéutico , Recombinación Genética , Factor A de Crecimiento Endotelial Vascular/metabolismo

RESUMEN

OBJECTIVE: To study the relationship between nucleotide excision repair gene ERCC1 and resistance to cisplatin in ovarian cancer. METHODS: The expression of gene ERCC1 in 58 ovarian cancer tissues and 4 cell lines were examined and its relationship with resistance to cisplatin were analyzed, the changes of sensitivity to cisplatin were observed after interference of ERCC1 gene with small interfering RNA (siRNA) in ovarian cancer cell lines. RESULTS: In 58 ovarian cancer tissues, the positive rate of ERCC1 protein in chemoresistant cases (57.89%) was higher than that in chemo-sensitive cases (28.21%, P = 0.029). The mRNA levels of ERCC1 gene in ovarian cancer cell lines ES-2, SKOV3, COC1, COC1/DDP were related to cisplatin IC50 values (r = 0.932, P <0.05). The sensitivity of cell lines ES-2, SKOV3, COC1/DDP cells to cisplatin was increased by 53.88, 5.07, and 3.75 times, respectively, after RNA interfering ERCC1 gene. CONCLUSION: ERCC1 gene is associated with the resistance to cisplatin and the sensitivity to cisplatin can be enhanced by RNA interfering ERCC1 in ovarian cancer.

Asunto(s)

Cisplatino/farmacología , Proteínas de Unión al ADN/metabolismo , Resistencia a Antineoplásicos , Endonucleasas/metabolismo , Neoplasias Ováricas/metabolismo , ARN Interferente Pequeño/genética , Adulto , Antineoplásicos/farmacología , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Reparación del ADN , Proteínas de Unión al ADN/genética , Endonucleasas/genética , Femenino , Humanos , Concentración 50 Inhibidora , Persona de Mediana Edad , Neoplasias Ováricas/genética , Neoplasias Ováricas/patología , Interferencia de ARN , ARN Mensajero/metabolismo , Transfección , Adulto Joven

RESUMEN

OBJECTIVE: To study the changes of DNA repair genes and enhanced anti-tumor effect of cisplatin induced by mifepristone in human ovarian cancer drug resistance cells. METHODS: The alterations of cisplatin concentration producing 50% inhibition (IC50 ) in the COC1/DDP cell lines were examined by methyl thiazolyl tetrazolium (MTT) assay. RT-PCR and flow cytometry were used to analyze the changes of the mRNA of ERCC1, BRCA1, hMLH1 genes and cell cycle and apoptosis. Subcutaneous implantation of COC1/DDP was established in nude mice and the enhanced anti-tumor effect of cisplatin by mifepristone was observed in vivo. RESULTS: Cisplatin IC50 values of COC1/DDP cell were decreased from (3.71 +/- 0.38) microg/ml to (3.18 +/- 0.46), (1.95 +/- 0.14), (0.64 +/- 0.18) microg/ml respectively when treated with 2.5, 5.0, 10.0 micromol/L mifepristone. Mifepristone could down-regulate the mRNA levels of ERCC1, BRCA1, hMLH1 genes and enhance G0/G1 phase block effect of cisplatin, and 2.5, 5.0, 10.0 micromol/L mifepristone combined with cisplatin increased rate of cell apoptosis from 0.08% to 5.11%, 9.13% and 12.24% respectively. The percentage of inhibition of xenograft tumor volume in combined treatment group was 70.1%, which was significantly different (P < 0.05). CONCLUSION: By down-regulating ERCC1, BRCA1, hMLH1 genes, blocking G0/G1 phase, and increasing apoptosis rate, mifepristone could enhance anti-tumor effect of cisplatin.

Asunto(s)

Apoptosis/efectos de los fármacos , Cisplatino/farmacología , Reparación del ADN , Mifepristona/farmacología , Neoplasias Ováricas/patología , Proteínas Adaptadoras Transductoras de Señales/genética , Proteínas Adaptadoras Transductoras de Señales/metabolismo , Animales , Antineoplásicos/farmacología , Proteína BRCA1/genética , Proteína BRCA1/metabolismo , Ciclo Celular/efectos de los fármacos , Línea Celular Tumoral , Cisplatino/administración & dosificación , Daño del ADN , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Modelos Animales de Enfermedad , Resistencia a Antineoplásicos , Ensayos de Selección de Medicamentos Antitumorales , Sinergismo Farmacológico , Endonucleasas/genética , Endonucleasas/metabolismo , Femenino , Humanos , Ratones , Ratones Desnudos , Mifepristona/administración & dosificación , Homólogo 1 de la Proteína MutL , Trasplante de Neoplasias , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Neoplasias Ováricas/genética , Neoplasias Ováricas/metabolismo , ARN Mensajero/genética , ARN Mensajero/metabolismo

RESUMEN

OBJECTIVE: To study changes of cisplatin sensitivity by RNA interfering the excision repair cross-complementing (ERCC) 1 gene in ovarian cancer cell lines. METHODS: The small interference RNA (siRNA) targeting ERCC1 gene was designed and synthesized by transcription in vitro, and transfected to ovarian cancer cell line ES-2. The mRNA and protein of ERCC1 were evaluated by means of RT-PCR, western blot and immunocytochemistry. The changes of cisplatin sensitivity after interference were examined by methyl thiazolyl tetrazolium (MTT) assay. RESULTS: In ES-2 cell, the mRNA and protein levels of ERCC1 were dramatically decreased 24, 48 and 72 hours after transfection. The sensitivity to cisplatin of ES-2 cell line was increased by 53.88 times after disturbing the ERCC1 gene. CONCLUSION: The sensitivity to cisplatin of ovarian cancer cell lines ES-2 could be enhanced by RNA interfering ERCC1 gene.

Asunto(s)

Cisplatino/farmacología , Proteínas de Unión al ADN/genética , Endonucleasas/genética , Interferencia de ARN , Antineoplásicos/farmacología , Western Blotting , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Supervivencia Celular/genética , Supervivencia Celular/fisiología , Reparación del ADN , Proteínas de Unión al ADN/metabolismo , Proteínas de Unión al ADN/fisiología , Relación Dosis-Respuesta a Droga , Resistencia a Antineoplásicos , Endonucleasas/metabolismo , Endonucleasas/fisiología , Femenino , Humanos , Neoplasias Ováricas/genética , Neoplasias Ováricas/patología , ARN Mensajero/genética , ARN Mensajero/metabolismo , ARN Interferente Pequeño/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transfección

RESUMEN

BACKGROUND: The purpose of the study was to evaluate the role of neoadjuvant chemotherapy and embolization via the anterior branches of the bilateral internal iliac arteries in treating patients with advanced ovarian epithelial carcinoma. METHODS: Forty-two patients with advanced ovarian epithelial carcinoma (study group) were treated via the anterior branches of the bilateral internal iliac arteries after cytoreductive surgery and 7 courses of adjuvant platinum-based combination chemotherapy. Primary cytoreductive surgery was performed in 43 patients with advanced ovarian epithelial carcinoma (control group), and then followed by 8 courses of adjuvant platinum-based combination chemotherapy. The rate of optimal cytoreductive surgery, survival rate, blood loss during operation and operative time were investigated in the two groups. Statistical significance was assessed using Student's t test, the Chi-square test and the log-rank test. RESULTS: In the study group, the rate of optimum debulking after platinum-based chemotherapy and embolization via the anterior branches of the bilateral internal iliac arteries was 71.43% (30/42) (chi(2) = 10.06, P < 0.005), and 9 (21.43%) of the 42 patients showed no gross residual disease after surgery. Blood loss and operative time were significantly decreased in the study group as compared with those in the control group (665.24 +/- 37.61 ml: 849.31 +/- 41.20 ml, t(1) = 33.21, P(1) < 0.001; 4.23 +/- 0.21 hours: 6.15 +/- 0.38 hours, t(2) = 28.92, P(2) < 0.01). In the study group, the mean survival time and the median overall survival were 33.66 months (95% CI, 24.73 to 42.58) and 26.00 months (95% CI, 19.22 to 32.78), respectively. The median disease-free interval was 18.20 months. In the control group, the mean survival time and the median overall survival were 32.38 months (95% CI, 24.92 to 39.84) and 25.00 months (95% CI, 22.80 to 27.20), respectively. The median disease-free interval was 14.20 months. The overall survival rates were not significantly different between the two groups (chi(2) = 6.48, P > 0.05). CONCLUSIONS: Neoadjuvant platinum-based combination chemotherapy and embolization via the anterior branches of the bilateral internal iliac arteries is an alternative treatment for patients with advanced ovarian epithelial carcinoma, in whom the chance of optimal cytoreductive surgery is low. The treatment can reduce blood loss, decrease operative time, and increase the rate of optimal cytoreductive surgery; but the median survival can't be improved significantly.

Asunto(s)

Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Embolización Terapéutica , Infusiones Intraarteriales , Neoplasias Glandulares y Epiteliales/terapia , Neoplasias Ováricas/terapia , Adulto , Anciano , Quimioterapia Adyuvante , Femenino , Humanos , Persona de Mediana Edad , Neoplasias Glandulares y Epiteliales/mortalidad , Neoplasias Ováricas/mortalidad , Tasa de Supervivencia

RESUMEN

OBJECTIVE: To study the effect of luteinized granulosa cell conditioned medium on cortical granule (CG) of the mouse oocytes matured in vitro. METHODS: Oocytes in germinal vesicle (GV) stage of Kunming mice were randomly divided into 2 groups according to different in vitro maturation (IVM) culture media. The study group medium contained 50% granulosa cell condition medium, follicle stimulating hormone 75 U/L and estrodial 1 nmol/L. The control group medium contained follicle stimulating hormone 75 U/L and estrodial 1 nmol/L. Oocytes were cultured for 16 or 18 hours. CG was examined by fluorescein isothiocyanate labeled Lens culinaris agglutinin under a confocal scanning laser microscope. RESULTS: After cultured for 16 hours, the nuclear maturation rates of control and study groups were 70.0% and 76.5%. After cultured for 18 hours, the maturation rates were 75.1% and 83.1%, respectively. There was no significant difference between the two groups. After cultured for 16 hours, there was no pronuclear formation in both groups. When culture was extended to 18 hours, fertilization occurred. After cultured for 16 hours, the rates of CGs forming a line under membrane were 10.0% and 50.0% in control and study groups respectively. When culture was extended to 18 hours, the rates rose to 57.1% and 91.6% accordingly. The rate of 18 h of each group was significantly higher than that of 16 h (both P < 0.001). The rate of study group of 18 h was significantly higher than that of control group (P < 0.05). CONCLUSION: Granulosa cell conditioned medium could improve the mouse oocytes maturation competence in vitro.

Asunto(s)

Gránulos Citoplasmáticos/metabolismo , Células de la Granulosa/metabolismo , Oocitos/citología , Animales , Técnicas de Cultivo de Célula , Núcleo Celular/metabolismo , Medios de Cultivo Condicionados/farmacología , Citoplasma/metabolismo , Gránulos Citoplasmáticos/efectos de los fármacos , Estradiol/farmacología , Femenino , Fertilización In Vitro , Hormona Folículo Estimulante/farmacología , Ratones , Microscopía Confocal

RESUMEN

OBJECTIVES: The purpose of this study was to determine whether overexpression of MDM2 could sensitize the ovarian cancer cell line A2780. METHODS: The wild-type p53-expressing cell line A2780 was stably transfected with pCMV-MDM2 (A2780-MDM2) or pCMV (A2780-V) as control. MTT assay and clonogenic survival assay were used to measure the cisplatin sensitivity. FACS and host cell (CAT) reactivation assay were used to estimate the change of cell cycle and ability of repairing cisplatin-induced DNA damage. RESULTS: Parental A2780 and A2780-V had similar cisplatin sensitivities, whereas A2780-MDM2 was two- to threefold more sensitive to cisplatin. Repair of cisplatin-induced DNA damage was reduced in A2780 cells overexpressing MDM2, compared to A2780 cells in which wild-type p53 function was intact. After cisplatin treatment, A2780-MDM2 cells showed a pronounced S-phase arrest; however, A2780 cells with intact wild-type p53 arrested primarily in G2/M phase. CONCLUSIONS: MDM2 overexpression can increase cisplatin cytotoxicity in A2780, with loss of G1/S checkpoint control and decreased cisplatin-DNA adduct repair. This suggests that ovarian cancers that overexpress MDM2 may be amenable to treatment with platinum compounds.

Asunto(s)

Proteínas Nucleares , Neoplasias Ováricas/genética , Proteínas Proto-Oncogénicas/genética , Antineoplásicos/farmacología , Western Blotting , Ciclo Celular/efectos de los fármacos , Ciclo Celular/genética , Cisplatino/farmacología , Daño del ADN , Reparación del ADN/genética , Femenino , Vectores Genéticos/genética , Humanos , Neoplasias Ováricas/tratamiento farmacológico , Neoplasias Ováricas/patología , Plásmidos/genética , Proteínas Proto-Oncogénicas c-mdm2 , Transfección , Células Tumorales Cultivadas