ABSTRACT
PURPOSE: We have previously demonstrated that 2-methoxyestradiol (2-ME) inhibits the growth of human endometrial cancer HEC-1-A and RL-95-2 cells in vitro. In this study, we examined the effects of 2-ME on human endometrial carcinoma in severe combined immune deficient (SCID) mice. The potential side effects of 2-ME on SCID mice were also investigated. METHODS: Severe combined immune deficient mice were injected with HEC-1-A cells (1 x 10(6)/mouse) and a 18 day administration of 2-ME was followed after 1 week cell implantation. Tumor volume, weight, body weight and blood chemistry were determined. Tumor tissues were examined with an antibody against the proliferative cell nuclear antigen (PCNA) and Ki-67. Liver, spleen, kidney, heart, lung and uterus were screened by pathological examinations. RESULTS: 2-ME (100 mg/kg p.o.) did not inhibit the growth of human endometrial carcinoma as compared to control. Necrotic areas were similar in both 2-ME-treated and -untreated tumor tissues. The expressions of PCNA and Ki-67 were similar in 2-ME-treated and untreated tumor sections. The wet weight of uterus was increased to more than threefold. The epithelial cells and glands in endometrium were increased. No significant difference was detected in blood AST, ALT and BUN. CONCLUSIONS: 2-ME has no antitumor effects on human endometrial carcinoma in our animal model. Its proliferative effects on endometrium and uterus might limit its use in gynecological cancers.
Subject(s)
Antineoplastic Agents, Hormonal/pharmacology , Endometrial Neoplasms/drug therapy , Estradiol/analogs & derivatives , 2-Methoxyestradiol , Animals , Antineoplastic Agents, Hormonal/administration & dosage , Cell Line, Tumor , Cell Proliferation/drug effects , Endometrial Neoplasms/pathology , Estradiol/administration & dosage , Estradiol/pharmacology , Female , Humans , Mice , Mice, SCID , Xenograft Model Antitumor AssaysABSTRACT
BACKGROUND: It has been demonstrated that 2-Methoxyestradiol (2-ME), one of the estrogen metabolites, induces apoptosis in many different tumor cell lines. In the present study, the effects of 2-ME on human cervical cancer HeLaS3 cells and on normal cervical epithelial cells were evaluated. MATERIALS AND METHODS: Acridine orange staining, DNA fragmentation arrays and flow cytometry were used to measure the apoptosis and cell cycle progression. In addition, the effect of 2-ME on expression of iNOS was measured by Western blot. RESULTS: 2-ME inhibited the growth of HeLaS3 cells. This growth inhibition was accompanied by apoptosis and G2/M cell cycle arrest. 2-ME increased the expression of iNOS in parallel with apoptosis. Moreover, apoptosis was prevented by the iNOS inhibitor 1400W. 2-ME treatment resulted in a slight increase of the G2/M population, but no apoptosis, in normal cervical epithelial cells. There was no synergetic effect between E2 and 2-ME. CONCLUSION: 2-ME induced apoptosis via the iNOS pathway and caused G2/M cell cycle arrest in human cervical cancer HeLaS3 cells, but showed only slight effects on normal cervical epithelial cells. These data suggest that 2-ME might be an adjuvant agent in the treatment of cervical cancer.
Subject(s)
Adenocarcinoma/drug therapy , Apoptosis/drug effects , Cell Cycle/drug effects , Estradiol/analogs & derivatives , Estradiol/pharmacology , Uterine Cervical Neoplasms/drug therapy , 2-Methoxyestradiol , Adenocarcinoma/pathology , Cell Division/drug effects , Cervix Uteri/cytology , Cervix Uteri/drug effects , Epithelial Cells/cytology , Epithelial Cells/drug effects , Female , G2 Phase/drug effects , HeLa Cells , Humans , Mitosis/drug effects , Uterine Cervical Neoplasms/pathologyABSTRACT
BACKGROUND: We investigated the effects of 2methoxyestradiol (2-ME), an endogenous estrogenic metabolite, on human endometrial cancer HEC-1-A and RL-95-2 cell lines. MATERIALS AND METHODS: After exposure of HEC-1-A and RL-95-2 cells to 2-ME, the morphological changes were evaluated by acridine orange staining and transmission electron microscopy. Cell cycle progress, apoptosis and necrosis were assessed by flow cytometry, DNA fragmentation and Western blot. RESULTS: 2-ME inhibited cell growth by blocking the S- and G2/M-phase in both cell lines, by inducing apoptosis in HEC-1-A cells and by causing necrosis in RL-95-2 cells. Apoptosis, on HEC-1-A cells, was accompanied by an increased expression of iNOS and STAT1. This apoptotic effect was prevented by the iNOS inhibitor 1400W and eliminated by the caspase inhibitor Z-VAD-FMK. Necrosis, on RL-95-2 cells, was due to a severe disruption of the mitochondrial membrane potential. 2-ME had no significant effect on normal human endometrial cells. CONCLUSION: The data suggest that 2-ME has an antitumor effect on human endometrial carcinoma cells (HEC-1-A and RL-95-2) and may contribute as a new therapeutic agent for endometrial cancer patients.
Subject(s)
Apoptosis/drug effects , Endometrial Neoplasms/drug therapy , Estradiol/analogs & derivatives , Estradiol/pharmacology , 2-Methoxyestradiol , Adult , Cell Cycle/drug effects , Cell Growth Processes/drug effects , Cell Line, Tumor , DNA-Binding Proteins/biosynthesis , Endometrial Neoplasms/metabolism , Endometrial Neoplasms/pathology , Endometrial Neoplasms/ultrastructure , Endometrium/cytology , Endometrium/drug effects , Female , Flow Cytometry , Humans , Intracellular Membranes/drug effects , Intracellular Membranes/physiology , Membrane Potentials/drug effects , Middle Aged , Mitochondria/drug effects , Mitochondria/physiology , Necrosis , Nitric Oxide Synthase/biosynthesis , Nitric Oxide Synthase Type II , STAT1 Transcription Factor , Trans-Activators/biosynthesisABSTRACT
Objetivos: avaliar variações de peso corporal, pressao arterial, glicemia em jejum, HbA1C, insulina, coleterol total, HDL-C, LDL-C, triglicérides, TGO, TGP, GGT e bilirrubina em mulheres usuárias de um implate único, subdérmico, de Silástico, contendo 55 mg (+10 por cento) de acetato de nomegestrol, durante dois anos. Métodos: dezoito voluntárias saudáveis e em idade reprodutiva, que desejavam fazer uso de anticoncepcionais e nao apresentavam contra-indicaçoes para o uso de contracepçao hormonal, participaram deste estudo. Todas as mulheres foram avaliadas antes do início do tratamento e a seguir, acompanhadas por um período de dois anos. Ao final do primeiro ano, as cápsulas foram retiradas e novas cápsulas foram inseridas. Resultados: o peso corporal aumentou de 54,9 + 1,5 kg na admissao para 55,3 + 2,0 Kg no 12º mês de uso (p<0,05), e para 56,0 + 2,7 Kg no 24º mês de uso. Registrou-se discreto aumento da pressao arterial, tanto sistólica quanto diastólica, no mês 12 (p<0,01). No mês 24, a pressao arterial nao era significativamente diferente dos valores de admissao. Todos os valores estiveram dentro dos limites da normalidade. Insulina, HbA1C, LDL-C e GGT permaneceram inalterados durante os vinte e quatro meses de uso do implante. Diminuiçao significativa do colesterol total (p<0,05) foi observada no 3º mês e de HDL-C (p<0,01) no 6º mês. Observou-se aumento significativo de triglicérides (p<0,05) apenas no 12º mês. Todas as alteraçoes de lipoproteínas foram inconsistentes, e os valores estiveram dentro dos limites da normalidade. Aumentos significativos dos níveis de glicemia em jejum (p<0,05 e p<0,01) foram observados respectivamente no 3º e no 6º mês. Diminuiçoes significativas da TGO (p<0,05, p<0,01 e p<0,05) foram observadas respectivamente no 6º, 18º e 24º mês e da TGP (p<0,05) no 18º mês. Somente se observou aumento significativo de bilirrubina (p<0,05) no 3º mês de uso do implante. Todas estas variaçoes permaneceram dentro dos limites da normalidade. Conclusoes: esses resultados demonstraram que, dentro dos limites da normalidade, as variaçoes de glicemia em jejum nao se correlacionaram às alteraçoes dos níveis de insulina. As alteraçoes discretas em lipoproteínas séricas, TGO, TGP e bilirrubina foram transitórias. Nao foram observados efeitos clínicos em lipoproteínas, metabolismo de carboidratos, níveis de insulina e funçao hepática entre as usuárias por dois anos.
