ABSTRACT
This study investigated the growth-regulating effects of progesterone (Prog) on nPR-negative malignant melanoma cells and the possible mechanisms. A375 and A875 cells were cultured and treated with Prog of different concentrations. For signal transduction pathway studies, the cells were pretreated with Prog receptor antagonist (RU486, 1x10(-7) mol/L) or MAPK inhibitor (U0126, 5x10(-6) mol/L) for 1 h and then co-incubated with prog (10(-9) mol/L) for another 24 h. Indirect immunofluorescence assay, MTT, flow cytometry and Western blotting were used for assessing the nPR expression, cell growth, cell apoptosis and ERK1/2 Phosphorylation, respectively. Our results showed that lower progesterone concentration promoted the proliferation of both A375 and A875 cells, but this growth-stimulatory effect decreased at progesterone concentration of 1x10(-7) mol/L or higher. The response could be abolished by MAPK inhibitor U0126, but could not be blocked by progesterone antagonist RU486. Flow cytometry exhibited that high concentration ([Symbol: see text]1 x 10(-7) mol/L) progesterone increased the apoptosis of the two cells in a dose-dependent manner. The level of ERK1/2 phosphorylation was increased by a lower progesterone concentration, but reduced by a higher concentration (1x10(-6) mol/L). These results suggest progesterone exerts growth-regulating effects on nPR-negative tumor cells through a non-genomic mechanism.
ABSTRACT
Objective To investigate the role of myeloid dendritic cells(mDC)in the pathogenesis of ankylosing spondylitis (AS),and to study the mechanism of tumour necrosis factor (TNF)-α blocker on the treatment of AS by counting the number of mDC before and after the treatment.Methods Peripheral blood from 21 AS patients treated with 50 mg rhTNFR-Fc and 15 AS patients treated with placebo were investigated at week 0,week 2 and week 6 in a randomized,double-blind,placebo-controlled trial.Three-color flow cytometry analysis was used to investigate the change of the number of mDC before and after the treatment.And their correlation with the clinical parameters was analyzed.Results MHC Class Ⅰ positive mDC(Lin-/CD11c+/HLA-abc+)in AS patients was slightly less than in healthy controls (not statistically significant).No significant change in MHC Class Ⅰ mDC number was observed after drug treatment.The number of MHC Class Ⅱ mDC cells was not correlated with clinical parameters.Conclusion The treatment with rhTNFR-Fc in AS induces a significant upregnlation of MHC-Ⅱ DC.
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Objective To study the relationship of symptoms of female gonococcal infections to Chlamydia trachomatis infection, serum sex hormone levels, etc. Methods A total of 136 gonorrhea female patients without obvious symptoms were recruited in this study together with 45 gonorrhea patients with obvious symptoms as the controls. Serum progesterone (P) and estradiol (E2) levels were measured by radio immunoassay (RIA). Cervical swabs were obtained from the subjects and eluted into isotonic saline solution, the elution was divided into 2 portions and tested for the levels of TNF-α and IL-1β by ELISA and for the DNA of C. Trachomatis and N. Gonorrhea with PCR. Statistical analysis was carried out by SPSS for Windows (version 12.0). Results There was no statistical correlation between C. Trachomatis infection and asymptomatic status of female gonococcal infection (χ2 = 0.016, P > 0.05). However, the decrease in the level of TNF-α and IL-1β significantly correlated with the increase in serum progestogen (r = -0.8798, -0.8935, respectively, both P < 0.01). Conclusion The high serum level of progesterone may be associated with the asymptomatic status of gonococcal infection.
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Objective To investigate the effects ofendothelin-1 (ET-1) and endothelin-3 (ET-3) on the expression of transformation growth factor-beta 1 (TGF-β1) and phosphorylation of Smad 3 in malignant melanoma cell line, A375. Methods Cultured A375 cells were classified into 5 groups, i.e. control group (no stimulation), ET-1 group (stimulated with ET-1), ET-1+BQ123 group (treated with ET-1 and BQ123),ET-1 + BQ788 group (treated with ET-1 and BQ788), ET-3 group (stimulated with ET-3), to receive different stimulation. The working concentrations were 0, 0.1, 1, 10, 100 nmol/L for ET-1 and ET-3, 10 μmol/L for BQ123 and BQ788. After another 12- and 24-hour culture, ELISA, RT-PCR and Western blot were used to detect the expression of TGF-β1 protein and mRNA as well as phosphorylated Smad 3 (P-Smad 3). Results The expression of TGF-β1 in A375 cells was up-regulated by ET-1, but down-regulated by ET-3, and both of the effects were in a concentration-dependent manner. Under the stimulation with ET-1 and ET-3 of 100 nmol/L, the level of TGF-β1 reached 1289.38 ± 89.42 ng/L per 105 cells and 85.09 ± 9.37 ng/L per 105 cells, respectively, significantly different from that in unstimulated cells (both P < 0.05). BQ123 signifi-cantly blocked the up-regnlatory effect of ET-1 on the expression TGF-β1 protein(P < 0.05), but BQ788 had no significant influence on the effect, so was the case with TGF-β1 mRNA. Western blot revealed that ET-1significantly elevated the expression of P-Smad 3 in A375 cells (P <0.05), and the elevation was significantly inhibited by BQ123, but not by BQ788. The expression of P-Smad 3 was statistically decreased by ET-3 in A375 cells (P <0.05). Conclusions The expression of TGF-β1 could be enhanced by ET-1, but suppressed by ET-3. It is likely that endothelin receptor A mediates the phosphorylation of Smad 3 induced by ET-1.
ABSTRACT
To investigate the role of progesterone receptor (PR) expression in malignant melanoma (MM), PR and proliferative cell nuclear antigen (PCNA) expression were immunohistochemistrically evaluated in a series of 35 specimens of MM, and the correlation between the immunohistochemistrical findings and clinicopathological data was also analyzed. PR expression was detected in 25.7% (9/35) of the patients with MM. No PR expression was observed in nevi. PR expression was inversely correlated with PCNA expression (r=-0.353, P=0.026). PR expression was slightly increased in females, subjects aged under 55 y, those with ulceration, non-acral subtype and diagnosis delay longer than 1 y, but the difference was not statistically significant. Selective expression of progesterone receptor in malignant melanoma might be correlated with inhibited tumor growth.