OBJECTIVETo explore the molecular mechanism of jiantai liquid (JTL) in improving endometrial receptivity of mice with embryo implantation dysfunction (EID).

METHODSMice model of EID induced by mifepristone were intervened with JTL (Twig of Chinese Taxillus, Red Sage root, Chinese Angelica, Milkvetch root, Chuanxiong rhizome), and sacrificed on day 8 of pregnancy. The endometrial estrogen receptor (ER) and progesterone receptor (PR) protein and their gene expressions were assessed by Western blot and semi-quantitative reverse transcriptase polymerase chain reaction (RT-PCR).

RESULTSLevels of ER and PR protein and their gene expressions in the JTL treated group were significantly higher than those in the model group respectively (all P < 0.05), and showed insignificant difference from those in the normal control group (all P > 0.05).

CONCLUSIONJTL could promote the development of endometrium and improve the embryo implantation by way of regulating the levels of ER and PR protein and gene expression in mice with EID.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Embryo Implantation ; drug effects ; Female ; Luteolytic Agents ; antagonists & inhibitors ; pharmacology ; Male ; Mice ; Mifepristone ; antagonists & inhibitors ; pharmacology ; Receptors, Estrogen ; biosynthesis ; genetics ; Receptors, Progesterone ; biosynthesis ; genetics ; Uterus ; metabolism

BACKGROUNDThe failure of endocrine treatment for advanced prostate cancer might be related to aberrant activation of androgen receptor (AR). Prostate cancer cell line LNCaP contains AR that can be activated by androgen, estrogen and progesterone. This study was set to investigate the effects of antisense AR RNA on growth of LNCaP cultured in medium containing varied concentrations of R1881, 17beta-estradiol, and progesterone, respectively.

METHODSLNCaP cells transfected with antisense AR RNA retroviral vector pL-AR-SN were designated as LNCaPas-AR. LNCaP cells containing empty vector pLXSN served as LNCaPNeo. LNCaP and LNCaPNeo were taken as controls. In vitro cell growth assay, proliferative cells of LNCaP and tranfected LNCaPs were counted by typan staining when they cultured with synthetic androgen R1881, 17beta-estradiol, and progesterone, respectively.

RESULTSGrowth of LNCaPas-AR was inhibited significantly (P < 0.05) compared with that of LNCaP and LNCaPNeo at 1 nmol/L R1881, 10 nmol/L 17beta-estradiol, and 1 nmol/L progesterone, respectively. No difference was seen between LNCaP and LNCaPNeo (P > 0.05). Microscopic observation showed that LNCaP and LNCaPNeo cells grew well, but only few LNCaPas-AR cells were alive.

CONCLUSIONSOur observations indicate that antisense AR RNA retroviral vector pL-AR-SN could change androgen-independent characteristics of LNCaP cells, which might shed some novel insights into the treatment of androgen-independent prostate cancer.

Androgen Receptor Antagonists ; Cell Division ; drug effects ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Estradiol ; pharmacology ; Humans ; Male ; Metribolone ; antagonists & inhibitors ; pharmacology ; Progesterone ; antagonists & inhibitors ; pharmacology ; Prostatic Neoplasms ; pathology ; therapy ; RNA, Antisense ; therapeutic use ; Receptors, Androgen ; genetics

OBJECTIVETo study the effect of BRCA1 in regulating the proliferation and migration of breast cancer cells stimulated by progesterone.

METHODSBreast cancer MCF-7 and T-47D cell were transfected with a vector containing the coding sequence of BRCA1 (pFlag-CMV2-BRCA1 wt) to induce BRCA1 overexpression or with the empty vector (control). The cells were then stimulated with progesterone, and the cell proliferation and migration were observed using MTT assay and wound healing assay, respectively. The proliferation and migration of MCF-7 cells were also observed following transfection with a small interfering RNA (siRNA) for BRCA1 knockdown or with a scrambled siRNA prior to progesterone stimulation.

RESULTSTransfection with the empty vector and with pFlag-CMV2-BRCA1 wt prior to progesterone stimulation caused significantly different proliferation rates in MCF-7 cells [(114.4∓6.0)% vs (82.1∓3.2)%, P<0.05] and in T-47D cells [(111.3∓4.3)% vs (84.2∓3.5)%, P<0.05], resulting also in significantly different cell migration rates (55.9% vs 15.8% in MCF-7 cells and 44.83% vs 10.43% in T-47D cells). Compared to the scrambled siRNA, BRCA1 siRNA transfection prior to progesterone stimulation significantly increased the proliferation rates [(114.4∓3.05)% vs (125.3∓4.0)%, P<0.05] and migration rate (39.2% vs 69.08%) of MCF-7 cells. The progesterone antagonist RU468 could antagonize the effects of BRCA1 knockdown in enhancing progesterone-stimulated MCF-7 cell proliferation and migration.

CONCLUSIONA decreased BRCA1 expression can enhance progesterone-stimulated tumor cell proliferation and migration in sporadic breast cancer.

BRCA1 Protein ; metabolism ; Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Female ; Genetic Vectors ; Humans ; Progesterone ; pharmacology ; RNA, Messenger ; genetics ; Receptors, Progesterone ; antagonists & inhibitors ; Transfection

OBJECTIVETo explore the mechanism of Jiantai liquid on the endometrium development of embryo implantation dysfunction mice.

METHODThe model of embryo implantation dysfunction mice was induced by mifepristone and treated by Jiantai liquid. All animals were sacrificed on day 8 of pregnancy. Estradiol and progesterone concentrations in serum and endometrium tissue homogenates were measured by radioimmunoassay method, the endometial expressions of estrogen receptor (ER)and progesterone receptor (PR)assessed by immunohistochemical SP method.

RESULTThere were no significantly differences in the estradiol and progesterone concentrations in serum and uterus tissue homogenates among three groups( P > 0.05). Absorbency and area rate of ER, PR in model group' s gland and stroma were higher than those in model group(P < 0.05), which was similar with the control group( P > 0.05).

CONCLUSIONJiantai liquid increase the implantation rate and improve the endometrial development by increasing the expressions of ER, PR in endometrium of embryo implantation dysfunction

Animals ; Astragalus membranaceus ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Embryo Implantation, Delayed ; drug effects ; Endometrium ; metabolism ; Female ; Loranthaceae ; chemistry ; Male ; Mice ; Mifepristone ; antagonists & inhibitors ; pharmacology ; Plants, Medicinal ; chemistry ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism ; Salvia miltiorrhiza ; chemistry

OBJECTIVETo observe and evaluate the phytoestrogenic effects and its mechanism of psoralen in estrogen receptor (ER) alpha and beta positive T47D and ishikawa cells.

METHODThe proliferation rate of T47D influenced by 1 x 10(-5) mol x L(-1) to 1 x 10(-9) mol x L(-1) psoralen and that of Ishikawa influenced by 1 x 10(-6) mol x L(-1) and 1 x 10(-7) mol x L(-1) psoralen were analyzed by MTT assay. PR mRNA expression in T47D was quantified by RT-PCR assay. Estrogen receptor antagonist ICI 182, 780 was employed as a tool. ER-alpha and ER-beta expression of T47D was measured by flow cytometry.

RESULTThe proliferation rates of T47D cells treated with 1 x 10(-5) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen and ishikawa cells treated with 1 x 10(-6) mol x L(-1) to 1 x 10(-7) mol x L(-1) psoralen were increased significantly. The RT-PCR result showed that 1 x 10(-7) mol x L(-1) and 1 x 10(-6) mol x L(-1) psoralen could increase PR expression in T47D cells. The above effects could be blocked by ICI 182,780. Psoralen could also induce the augment of ER-alpha and ER-beta expression in T47D cells significantly.

CONCLUSIONPsoralen has phytoestrogenic effects. The effects are attained through ER pathway.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Estradiol ; analogs & derivatives ; pharmacology ; Female ; Ficusin ; pharmacology ; Flow Cytometry ; Humans ; Receptors, Estrogen ; antagonists & inhibitors ; genetics ; Receptors, Progesterone ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

Insulin resistance (IR) in dogs is suspected when hyperglycemia is present despite administration of insulin doses greater than 1.0 to 1.5 UI/kg. IR is caused by increases in counter regulatory hormones concentrations (glucagon, glucocorticoids, catecholamines and growth hormone). This study was conducted to investigate the use of aglepristone (RU 46534), a P4 receptor antagonist, for the treatment of IR diabetes mellitus in bitches during the luteal phase. All animals were treated with porcine insulin zinc suspension (Caninsulin) and aglepristone (Alizin) 10 mg/kg subcutaneously at day 1, 2, 9 and 17 from diagnosis. At day 5, no significant variation in glycemia was shown. At day 12 and 20, serum glucose concentrations were significant lower (p < 0.05). From day 12 the insulin dose was reduced to 0.8 IU BID. Insulin was reduced in the following weeks and glycemia was controlled.
Animals ; Blood Glucose/analysis ; Diabetes Mellitus/drug therapy/etiology/*veterinary ; Dog Diseases/*drug therapy/etiology ; Dogs ; Estrenes/*therapeutic use ; Estrous Cycle ; Female ; Hypoglycemic Agents/therapeutic use ; Insulin Resistance ; Pregnancy ; Radioimmunoassay/veterinary ; Receptors, Progesterone/*antagonists & inhibitors

OBJECTIVE: To observe the expression of progesterone receptor (PR), interleukin-1β (IL-1β), and cyclooxygenase-2 (COX-2) induced by lipopolysaccharide (LPS) or Toll-like receptor 4 antagonist (TLR4 mAb) in decidual cells in vitro, and then to explore the effect of LPS and its antagonist on PR of decidual cells and the relation between PR and inflammatory cytokines. METHODS: We isolated and cultured human decidua of early abortion in the sterile state. When the cells passaged to the 4th generation, the cells were randomly divided into 6 pore plates: A control group was added the culture medium alone; experimental group I was added 100 ng/mL of LPS; experimental group II was add 1 μg/mL of TLR4 mAb; experimental group III was added 3 μg/ mL of TLR4 mAb; experimental group IV was added 1 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS; and experimental group V was added 3 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS for 24 h culture. Subsequently, HE staining and immunofluorescence were used to observe the morphology and identify the purity of decidual cells in the 6 groups. The levels of mRNA expression of PR, IL-1β, and COX-2 were detected by reverse transcription PCR (RT-PCR). RESULTS: LPS reduced the mRNA expression of PR (P<0.05), increased the mRNA expression of IL-1β and COX-2 (P<0.05). TLR4 mAb increased the mRNA expression of PR (P<0.05) and reduced the mRNA expression of IL-1β (P<0.05) after LPS-stimulated decidual cells. High concentrations of TLR4 mAb reduced the mRNA expression of COX-2 (P<0.05) after LPS stimulated decidual cells. CONCLUSION The mRNA expression of PR is reduced, and the mRNA expressions of IL-1β and COX-2 are increased after LPS-stimulated decidual cells in vitro. TLR4 mAb antagonize the role of LPS on PR, IL-1β, and COX-2.
Adult ; Cells, Cultured ; Cyclooxygenase 2 ; genetics ; metabolism ; Decidua ; cytology ; metabolism ; Female ; Humans ; Interleukin-1beta ; genetics ; metabolism ; Lipopolysaccharides ; pharmacology ; RNA, Messenger ; genetics ; metabolism ; Receptors, Progesterone ; genetics ; metabolism ; Toll-Like Receptor 4 ; antagonists & inhibitors ; Young Adult

OBJECTIVETo explore the estrogenic activity of ethanol extract from adzuki bean Phaseolus angularis and its effect on progesterone receptor (PR) level of human breast cancer MCF-7 cells.

METHODThe modified MCF-7 cell proliferation assay was used to evaluate the estrogenic activity of adzuki bean extract. And its effect on PR mRNA and protein were addressed using RT-PCR measurements and western blotting analysis respectively in which pure estrogen receptor antagonist ICI 182,780 was employed as the tool.

RESULTThe estrogenic activity of adzuki bean extract at various concentrations (10-200 microg x mL(-1)), expressed as proliferative effect (PE) relative to that of solvent control, was examined. The results indicated that the extract of adzuki bean was able to induce MCF-7 cell growth with a maximum at 100 microg x mL(-1). The results of RT-PCR and western blotting showed that PR mRNA and protein could be significantly induced by adzuki bean extract in MCF-7 cells. Moreover, the specific estrogen receptor antagonist ICI 182,780 could block these reactions.

CONCLUSIONEthanol extract of adzuki bean has the estrogen-like activities through the estrogen response pathway.

Breast Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Estradiol ; analogs & derivatives ; pharmacology ; Estrogen Antagonists ; pharmacology ; Female ; Humans ; Phaseolus ; chemistry ; Phytoestrogens ; isolation & purification ; pharmacology ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Progesterone ; biosynthesis ; genetics ; Seeds ; chemistry

OBJECTIVETo investigate the expression of ER alpha in chemically induced, ER alpha-negative human breast cancer MDA-MB-435 cells and its restoration of the responsiveness to endocrine therapy.

METHODSMDA-MB-435 cells were treated with HDAC inhibitor trichostatin A(TSA)and DNMT1 inhibitor 5-AZA-CdR (AZA). The mRNA level of ER alpha, PR and PS2 in treated MDA-MB-435 cells was detected by RT-PCR. The WST-8 (water-soluble tetrazolium salt-8) method was used to analyze the proliferation rate of the cells. Xenograft in female nude mice was used to further explore the change of proliferation rate of treated MDA-MB-435 cells in vivo.

RESULTSAfter treatment with AZA and TSA, mRNA expression of ER alpha, PR and pS2 was up-regulated in MDA-MB-435 cells. The mRNA level of ER alpha was the hightest when MDA-MB-435 cells were treated with 2.5 micromol/L AZA and 100 ng/ml TSA. The treated MDA-MB-435 cells showed different proliferation rate in various media containing different concentration of estrodial. The MDA-MB-435 cells showed down-regulated proliferation rate after treatment with the combination of 2.5 micromol/L AZA and 100 ng/ml TSA, and 4-OH tamoxifen could suppress the growth rate of the induced MD-MBA-435 cells but not the untreated cells. The treated MDA-MB-435 cells showed slower proliferation rate than that of untreated cells in vivo (P <0. 01), and the proliferation rate of the treated MDA-MB-435 cells became lower when the nude mice were deprived of estrogen by castration (P <0. 01).

CONCLUSIONAfter treatment with TSA and AZA, ER alpha-negative MDA-MB-435 cells can express functional ER alpha and regain responsiveness to estrogen both in vitro and in vivo. HDAC inhibitor and DNMT1 inhibitor may play an important role in restoration of sensitivity of ER alpha-negative breast cancers to endocrine therapy.

Animals ; Azacitidine ; analogs & derivatives ; pharmacology ; Breast Neoplasms ; genetics ; pathology ; prevention & control ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; DNA Modification Methylases ; antagonists & inhibitors ; Enzyme Inhibitors ; pharmacology ; Estrogen Receptor alpha ; genetics ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Histone Deacetylase Inhibitors ; Humans ; Hydroxamic Acids ; pharmacology ; Mammary Neoplasms, Experimental ; genetics ; pathology ; prevention & control ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Ovariectomy ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Progesterone ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Trefoil Factor-1 ; Tumor Suppressor Proteins ; genetics ; Xenograft Model Antitumor Assays