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Objective To explore the mechanism resistance of medroxyprogesterone 17-acetate (MPA) on the endometrial cancer side-population (SP) cells.Methods (1) Ishikawa-SP cells from endometrial cancer cell lines Ishikawa were be separated by Hoechst 33342 dyeing method and flow cytometry analysis.The clone formation efficiency between Ishikawa-SP cells and Ishikawa-non-SP cells were performed by clone formation assay.Breast cancer resistance protein (BCRP) was examined by immunocytochemistry method.(2)Ishikawa,Ishikawa-SP,Ishikawa-non-SP cells were treated with various concentrations of MPA at 5,10,15,20 μmoL/L.After cultured for 24,48,and 72 hours,cells growth were measured by methanethiosulfomate(MTS) assay.(3) The groups of Ishikawa,Ishikawa-SP,Ishikawa-non-SP cells incubated with MPA at the half maximal inhibitory concentration(IC50) were selected for cell apoptosis assay by using flow cytometry.After MPA treatment,the expression of caspase-3 was examined by immunocytochemistry method.Results (1)There were few proportion of Ishikawa-SP cells in Ishikawa endometrial carcinoma,which were 2.7％.There were stronger clone formation efficiency for Ishikawa-SP cells than that for Ishikawa-non-SP cells in Ishikawa [(6.02 ± 1.17)％ vs.(0.53 ±0.20)％,P =0.001].And there were higher level expression of BCRP (P =0.001)and also more resistant Taxol and radiation between Ishikawa-SP cells and Ishikawa-non-SP cells.(2)The inhibitory effect of MPA was concentrationdependent and time-dependent.(3)After MPA treatment,the apoptosis rates of Ishikawa-SP,Ishikawa-nonSP,Ishikawa were (4.01 ± 0.43) ％,(9.30 ± 0.67) ％,and (4.64 ± 0.18) ％,respectively (P ＜ 0.05).The level expression of caspase-3 in Ishikawa group after MPA treated were higher than that in Ishikawa-SP group.Conclusion MPA may be inhibit the growth of endometrial cancer,Ishikawa-SP and Ishikawa-nonSP cells,while Ishikawa-SP may be more resistant to MPA than Ishikawa-non-SP,which mechanism of resistance on MPA may be related to the properties of cancer stem-like cells and cell apoptosis.
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OBJETIVO: Avaliar as alterações histomorfométricas nas mamas de ratas tratadas com estrogênio e/ou progestagênio por curto período de tempo. MÉTODOS: Foram divididas em quatro grupos 40 ratas ooforectomizadas: GC-recebeu veículo; GE-recebeu benzoato de estradiol (37,6 µg/animal); GP-recebeu acetato de medroxiprogesterona (11,28 mg/animal) e, GEP-recebeu benzoato de estradiol (37,6 µg/animal) e acetato de medroxiprogesterona (11,28 mg/animal). No grupo GE, o estradiol foi administrado durante sete dias, por via subcutânea. Já no grupo EP o estradiol foi administrado nos primeiros sete dias e o progestagênio por mais 23 dias, por via subcutânea. Vinte e quatro horas após a última administração dos hormônios, os animais foram anestesiados e o primeiro par de mamas inguinais removido, imerso em formaldeído a 10 por cento e processado para inclusão em parafina, sendo os cortes corados pela Hematoxilina-Eosina. Foram avaliadas a morfologia e a área ocupada pelo parênquima mamário, sendo os dados submetidos à análise de variância complementado pelo teste de Kruskal-Wallis (p < 0,05). RESULTADOS: As mamas no grupo-controle apresentaram-se atrofiadas, sendo que, nos animais dos grupos GE e GEP, nota-se a presença de alvéolos típicos contendo secreção no seu interior, já nos animais tratados somente com progestagênio (GP) notam-se alvéolos formados por células volumosas que ocupam praticamente todo o lúmen alveolar. A morfometria mostrou haver maior área de parênquima mamário nos animais tratados com hormônios (GE = GP > GEP > GC; p < 0,05) CONCLUSÃO: O estradiol e o progestagênio apresentaram efeito proliferativo no parênquima mamário. No entanto, a administração prévia de estradiol modifica a ação do progestagênio no tecido mamário da rata.
OBJECTIVE: To evaluate the breast histomorphometric changes in rats treated with estrogen and/or progestogen for a short period of time. METHODS: Forty oophorectomized rats were divided into four groups: GC, vehicle; GE, treated with estradiol benzoate (37.6 mg/animal); GP, treated with medroxyprogesterone acetate (11.2 mg/animal) and GEP, treated with estradiol benzoate (37.6 mg/animal) plus medroxyprogesterone acetate (11.28 mg/animal). In GE group, estradiol was administered subcutaneously for seven days; in GEP group, estradiol was administered once in a day for the first seven days and the progestogen over the next 23 days both subcutaneously. Twenty-four hours after the last hormone administration, the animals were killed upon deep anesthesia and the first inguinal breasts were removed, fixed in 10 percent formaldehyde and processed to be included in paraffin, with the sections being stained by hematoxylin-eosin. Morphology and the area occupied by mammary parenchyma were assessed, with the data undergoing analysis of variance followed by the Kruskal-Wallis test (p < 0.05). RESULTS: The control group breasts were found atrophic and, in GE and GEP group animals, typical alveoli with secretion inside are present; in progestogen-treated animals (GP), alveoli formed by large cells occupying almost the entire alveolar lumen are noted. Morphometric analysis showed a larger mammary parenchyma area in hormone-treated animals (GE = GP > GEP > GC; p < 0.05). CONCLUSION: Estradiol and progestogen had a proliferative effect on mammary parenchyma. However, prior estradiol administration changes the progestogen action on rat mammary tissue.
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Animals , Female , Rats , Estradiol/pharmacology , Estrogen Replacement Therapy , Mammary Glands, Animal/drug effects , Medroxyprogesterone Acetate/pharmacology , Progestins/pharmacology , Estradiol/analogs & derivatives , Mammary Glands, Animal/pathology , Ovariectomy , Random Allocation , Rats, WistarABSTRACT
Objective To study the effect of 17α-hydroxyprogesterone caproate (17P) and medroxyprogesterone acetate (MPA) on expression of tumor necrosis factor-α (TNF-α)and cyclooxygenase-2 (COX-2) in placenta and uterine myometrium of inflammation-induced preterm birth mouse model to investigate the mechanism of preventing inflammation-induced preterm birth by progestogen.Methods Thirty clean CD-1 mice were divided into 6 groups (5 mice in each group) at 15th day of gestation:control group,lipopolysaccharides (LPS) group,17P 1 mg+LPS group,17P 2 mg+ LPS group,MPA 1 mg+LPS group and dimethyl sulfoxide (DMSO) + LPS group.Progestogens at different dosage were administered 1 h before LPS and 6 h after LPS administration.After these mice were sacrificed,TNF-α and COX-2 levels in the myometrium and placenta were detected by real-time PCR and immunohistochemistry.Data were analyzed by ANOVA,and comparisons between groups were adopted LSD method.Results 1.The comparison of relative expression of COX-2 mRNA and TNF-α mRNA in myometrium and placenta among groups:(1) Expressions of COX-2 mRNA and TNF-α mRNA in myometrium and placenta in the study groups were obviously higher than those of control group (P＜0.05).(2) COX-2 mRNA expression in myometrium of 17P 1 mg+LPS group (11.410±3.931),17P 2 mg+LPS group (8.352±3.209) and MPA 1 mg+LPS group (11.920± 2.905) were obviously lower than that of LPS group (20.540± 4.147) and DMSO+ LPS group (18.620 ± 4.156) (P＜0.05,respectively) ; although TNF-α mRNAexpression had similar trends among these groups,there were no statistical significance (P＞0.05,respectively).(3) COX-2 mRNA expression in placenta of 17P 1 mg+LPS group (10.864±3.777),17P 2 mg+LPS group (7.084±1.667) and MPA 1 mg+LPS group (11.830±3.652) were obviously lower than that of LPS group (18.920±4.106) and DMSO+LPS group (23.820±7.554)(P＜0.05,respectively).(4) TNF-α mRNA expression in placenta in 17P 1 mg+LPS group (14.340±1.618),17P 2 mg+ LPS group (11.488 ± 2.910) and MPA 1 mg+ LPS group (13.040 ± 2.982) were obviously lower than that of LPS group (24.240±7.059) and DMSO+LPS group (23.040±5.896) (P＜0.05,respectively).2.The comparison of protein expression of COX-2 and TNF-α in placenta among groups:(1) The expression of COX-2 protein in placenta of the study groups was significantly higher than that of control group (P＜0.05).(2) There were no differences among the COX-2 protein expression of 17P 1 mg+ LPS group (14 360.92± 1766.01),17P 2 mg+ LPS group (13 340.18±965.35) and MPA 1 mg+LPS group (12 870.81±1521.97)(P＞0.05),while the COX-2 protein expressions of them were significantly lower than that of LPS group (16 426.64 ± 1823.87) and DMSO+LPS group (16 761.23±2388.17)(P＜0.05,respectively).(3) There were no differences among the TNF-α protein expression of 17P 1 mg+LPS group (22 750.96±4656.68),17P 2 mg+LPS group (22766.24± 3500.34) and MPA 1 mg+LPS group (20770.01±3318.48)(P＞0.05),while the TNF-α protein expressions of them were significantly lower than that of LPS group (26204.49±5090.34) and DMSO+LPS group (27346.18±3269.30)(P＜0.05,respectively).Conclusions 17P and MPA might prevent the preterm parturition of inflammation-induced mouse model by inhibiting inflammation cytokines and prostaglandins.
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Objective To evaluate effects and safety of gonadotrophin-releasing hormone agonist (GnRH-a) combined with transdermal estradiol and medroxyprogesterone acetate in the treatment of endometriosis. Methods From January I st, 2007 to July 31 st, 2007, 28 endometriosis patients underwent laparnscopic or transabdominal surgery in Obstetrics and Gynecology Hospital affiliated to Fudan University were randomly divided into group A and group B. 14 patients in group A received 3.6 mg goserelin once every 4 weeks, 12 weeks in all 14 patients in group B received goserelin and added 1/2 piece of half-hydrate estradiol every week and 6 mg oral medroxyprogesterone acetate per day, 12 weeks in all. Serum estradiol (E2 ), follicle stimulating hormone(FSH), bone gla protein levels, visual analogue scale (VAS) of pain, bone mineral density of lumbar spine, vaginal exfoliate cell spurs and the form of Kupperman were compared in patients before and after treatment. Results (1 ) After treatment, the level of FSH and E2levels were (5.0 ± 2. 6 ) U/L and (29 ± 17 ) pmol/L in group A and (3.0 ± 1.5 ) U/L, and (87 ± 53 ) pmol/L in group B, which were significantly lower than those before treatment [FSH (17. 0 ± 12. 2) U/L, and E2 (184 ± 194) pmol/L in group A and FSH :(15.3±13.6)U/L and E2: (281±242) pmol/L in group B, P ＜ 0. 01]. On the seventh day after three-month GnRH-a treatment, it was observed that the level of E2 was higher and FSH was lower in group B than the level of E2 and FSH of group A (P ＜ 0. 01 ). (2 ) After treatment, the basal vaginal exfoliate cell proportion in group A [(66. 2 ± 29. 0) %] was significantly lower than that in group B [(11.8 ± 28. 0) %, P ＜ 0. 01] ; while patients in group A owned a lower proportion of the middle [(29. 1 ± 23.1 ) %], superficial layers [(4. 0 ± 5.5 ) %] and esinophilic cells [(2. 3 ± 2. 6)%]than patients group B [middle layer: (73. 0 ± 25.2)% ; superficial layer: (15. 2 ± 10. 9)% ; esinophilic cells: (10. 8 ± 7.9 ) % ; P ＜ 0. 01]. (3) Before the treatment, patients' VAS scores of total, pelvic pain, dysmenorrheal and dyspareunia were 7.43±3. 20,2. 35 ± 1.82, 4. 93 ± 1.98 and 0. 14±0. 53 in group A and were 7.71±2. 02, 2. 57 ± 1.60, 4. 86 ± 1.56 and 0. 29 ± 1.07 in group B; after treatment, the scores above were changed to 0. 14±0. 36,0. 07±0. 27,0. 07±0. 27and 0 in group A and 0. 36±0. 50, 0. 29±0. 47, 0. 07±0. 27 and 0 in group B, which were all significantly lower than those before treatment separately (P ＜0. 01 ). When menstruation recovered, the scores were 0. 21±0. 43, 0. 07±0. 27, 0. 14 ± 0. 36, and 0 in group A and 0. 50±0. 65, 0. 29±0. 47, 0. 21±0. 43 and 0 in group B, which were also significantly lower than those before treatment (P ＜ 0.01 ), however, no statistical difference was found between groups at any time spot(P ＞ 0. 05). (4) In group A, the bone density after treatment [(0. 96 ± 0. 06 ) g/cm2] was lower than that before treatment [(0. 99 ± 0. 06 ) g/cm2, P ＜ 0.01 )]. In group B, the index was (0. 98 ± 0. 09) g/cm2, which was lower than that before treatment [(0. 99 ± 0. 10 ) g/cm2, P = 0. 201]. No statistical difference was found between groups(P ＞ 0. 05 ). The bone loss rate were (- 2. 77 ± 1.97 ) % in group A and (- 0. 93 ± 2. 86 ) % in group B (P = 0. 058 ). Before treatment, the bone gla protein was (13±3) μg/L in group A and (13±6) μg/L in group B. After treatment, the bone gla protein levels was (17±6)μg/L in group A, which was higher than that before treatment (P ＜ 0. 01 ), the level was (16±6)μg/L in group B, which was higher than that before treatment, however showed no statistical difference(P =0. 053). No difference was found in bone gla protein before and after treatment between two groups (P＞0. 05). (5) The form of Kupperman after treatment were 15±7 in group A and 11±6 in group B, which did not show significant difference (P ＞ 0. 05 ). The incidence of flash and sweat were 93% (13/14)in group A, which was significantly higher than that 57% (8/14) in group B(P ＜0.01 ). Conclusion The add-back therapy that consists of an estradiol patch and oral medroxyprogesterone acetate is effective and safe treatment for endometriosis.
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Objetivo: O objetivo deste estudo foi avaliar em mulheres com tumores císticos de ovário, a proporção que, somente com o tratamento clínico e/ou punção, não necessitaram de cirurgia; a influência da medicação e das doenças associadas na indicação de cirurgia e se a punção reduziu significativamente o tamanho dos cistos. Casuística e métodos: Selecionaram-se 71 mulheres com idade entre 19 e 70 anos de idade, portadoras de tumores císticos de ovário maior que 5 cm, com características de benignidade ao ultra-som transvaginal com Doppler colorido e pulsado e com perfil endócrino e marcadores séricos tumorais normais. Foram divididas em cinco grupos: 1) Grupo A: 15 mulheres que fizeram uso de acetato de noretisterona; 2) Grupo B: 13 mulheres que fizeram uso de acetato de medroxiprogesterona; 3) Grupo C: 14 mulheres que utilizaram contraceptivo oral; 4) Grupo D: 15 mulheres que foram tratadas com análogo de GnRH e 5) Grupo E: 14 mulheres que não fizeram uso de medicação. Todos os grupos foram acompanhados por um ano. No final do primeiro trimestre, se o cisto persistiu, foi realizada a punção. No final do segundo trimestre, se houve recidiva do cisto, foi indicada cirurgia. As pacientes tiveram alta após um ano de acompanhamento com o tratamento do cisto concluído. Resultados: Não houve diferenças significantes entre os cinco grupos. Constatou-se que a condição de uso de medicação para doença associada teve influência significativa no resultado dos tratamentos. Verificou-se que a redução do tamanho do cisto com a punção foi efetiva após 9 meses de acompanhamento. Observou-se que 7% das mulheres tiveram indicação direta para cirurgia sem a punção, portanto, a proporção de recidiva da punção foi de 19,3% enquanto a porcentagem de mulheres que não fizeram a cirurgia foi de 73,2%. Conclusão: A proporção de mulheres que responderam adequadamente somente com o tratamento clínico e/ou a punção, não necessitando pois de cirurgia foi de 73,2%...
Objectives: To evaluate the rate of women with ovarian cystic tumors submitted to diferent regimens of hormonal treatment and/or aspiration that didnt need surgery. To access the effectiveness of ovarian cyst aspiration guided by ultrasonography (USG) and to study the influence of medication and associated diseases in the need for surgery. Patients and Methods: Seventy one (71) women, in an age range from 19 to 70 years, diagnosed with ovarian cysts larger than 5 cm in length, with benign aspect, examined using an ultrasonographic transvaginal probe and color Doppler. All the women have been submitted to measurements of ser um tumor markers and hormonal levels. They were grouped in five different categories: 1) Group A: 15 women treated with noretisterone acetate; 2) Group B: 13 subjects treated with medrooxiprogesterone acetate; 3) Group C: 14 patients treated with combined oral contraceptive; 4) Group D: 15 women treated with GnRH analog, and 5) Group E: 15 women with no treatment control). After 3 months, all the patients were reevaluated. In those ones whose cysts remained, cyst aspiration guided by ultrasonography was indicated. After 3 extra months, the women were submitted to a new USG examination and in those whose cysts still remained, surgery was performed. All the patients were followed for one year. Results: There was no significant difference among the groups of hormonal treatment. The use of medication promoted a significant influence in the treatment outcomes. We also noted that the cyst aspiration was effective in reducing the size of the cysts after 9 months of follow up. Seven per cent (7%) of the patients have had immediate indication for surgery. Among those who was submitted to cyst aspiration, 19.3% had to be resubmitted to treatment because of re-incidence of the cyst, whereas, 73.2% avoid the need for surgery. Conclusions: The proportion of subjects treated with hormones and/or cyst aspiration, with no need for surgery, was of 73.2%...