Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometri...Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometrial stromal cells separated from the proliferative endometrial tissues were incubated with medium alone, 17-β estradiol （E2,10^-8 mol/L）, medroxyprogesterone acetate （MPA, 10^-6 mol/L）, E2（10^-8 mol/L）＋MPA （10^-6 mol/L）, E2 （10^-8 mol/L）＋MPA （10^-6 mol/L）＋RU486 （10^-5 mol/L） or HB-EGF （10 ng/ml） for 48 h respectively. The expressions of MMP-9 and TIMP-1 were detected by in situ hybridization, immunocytochemistry, reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting. Results Compared with control group [mRNA, 0. 729 ± 0. 090 （MMP-9） and 1.056± 0.154 （TIMP-1）; protein, 0.545 ±0.086 （MMP-9） and 0.745 ±0.154 （TIMP-1）], expressions of MMP-9 and TIMP-1 in E2 alone, progestin alone or E2 combined with progestin group were respectively：mRNA, 0.413 ± 0.069, 0.402 ± 0.073 and 0.407 ± 0.039; 0.487 ± 0.093, 0.503 ± 0.093 and 0.468 ± 0.075：protein, 0.294 ± 0.076, 0.331 ±0.064 and 0.265 ±0.049; 0.425 ±0.085, 0.397 ±0.065 and 0.435 ± 0.099. RU486 weakened the expression level of down-regulation, while HB-EGF elevated the level of MMP-9 and TIMP-1 after 48 h treatment （mRNA, 0.955 ± 0.068 and 1.396 ± 0.238; protein, 0. 780 ± 0.109 and 0.985 ± 0.165）. Conclusions 1） Both E2 and progestin can down-regulate the expressions of MMP-9 and TIMP-1 in endometrial stromal cells, but RU486 can inhibit the effect. 2） HB-EGF can elevate the level of MMP-9 and TIMP-1. 3） E2, progestin and HB-EGF have effect on the ratio of MM-P/TIMP-1.展开更多
AIM： To evaluate the effects of angiopoietin-1 （Ang-1） on adhesion of gastric cancer cell line BGC-823 and expression of integrin β1, CD44V6, urokinase-type plasminogen activator （uPA） and matrix metalloproteina...AIM： To evaluate the effects of angiopoietin-1 （Ang-1） on adhesion of gastric cancer cell line BGC-823 and expression of integrin β1, CD44V6, urokinase-type plasminogen activator （uPA） and matrix metalloproteinase-2 （MMP-2）. METHODS： BGC-823 cells were transfected transiently with adenovirus-Ang-1 （Ad-Ang-1）. Cells transfected transiently with adenovirus-green fluorescent protein （Ad-GFP） and untransfected cells were used as a negative and blank control group, respectively. The cell adhesion rate between cell and extracellular matrix （ECM） was determined by cell adhesion assay. To investigate whether Ang-1 could reinforce gastric carcinoma metastasis, we performed migration and invasion assays in BGC-823 cells. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 were detected by reverse transcription polymerase chain reaction and Western blotting, respectively. The expression of integrin β1 and CD44V6 was measured by immunohistochemistry. RESULTS： BGC-823 cells were transfected successfully. The adhesion rate increased significantly in the Ad-Ang-1 group （P 〈 0.05）. The Ad-Ang-1-transfected group had a significant increase in migration and invasion compared with that of the mock-transfected and Ad-GFP groups. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 in the Ad- Ang-1 group was higher than that in the Ad-GFP and blank control groups （P 〈 0.05）. Compared with mocktransfected and Ad-GFP groups, integrin 131 and CD44V6 expression intensity greatly increased （P 〈 0.05）. CONCLUSION： Transfection of Ang-1 into human gastric cancer cell line BGC-823 can significantly increase expression of integrin β1 and CD44V6, by which cell adhesion and metastasis to the ECM are promoted.展开更多
Objective To investigate the role of matrix metalloproteinase-1 （MMP-1）/protease- activated receptor-1 （PAR-l） signaling in the cervical cancer invasion. Methods RT-PCR was used to test the mRNA level of MMP-1 in ...Objective To investigate the role of matrix metalloproteinase-1 （MMP-1）/protease- activated receptor-1 （PAR-l） signaling in the cervical cancer invasion. Methods RT-PCR was used to test the mRNA level of MMP-1 in 59 cases of cervical squamous cell cancer. Cell invasion was evaluated by Transwell assay to explore the effect of adding human recombinant MMP-1 （rhMMP-1） and PARI-siRNA on cervical cancer invasion. Results In cervical cancer tissues, more MMP-1 expression was observed than that in the normal cervical tissues, and its expression correlated with cancer status. Human recombinant MMP-1 （rhMMP-1） could promote Hela cell invasion, and its number of invasive cell correlated with the concentration of rhMMP-1. Disrupting the expression of PAR-1 reduced the MMP-1 promoting-effect on Hela cell invasion, but had no effect on non-MMP-1 invasive action. Conclusion The MMP-1/PAR-1 signaling pathway is involved in cervical cancer invasion. Therefore, blocking PAR-1 may represent a new therapeutic option for metastatic cervical cancer.展开更多
目的研究活动期类风湿关节炎(rheum atoid arthritis,RA)患者血清基质金属蛋白酶-3(m atrix m etalloproteinase3,M M P-3)、金属蛋白酶组织抑制剂-1(tissue inhibitors ofm etalloproteinase1,TIM P-1)的水平及其相关影响因素,探讨M M ...目的研究活动期类风湿关节炎(rheum atoid arthritis,RA)患者血清基质金属蛋白酶-3(m atrix m etalloproteinase3,M M P-3)、金属蛋白酶组织抑制剂-1(tissue inhibitors ofm etalloproteinase1,TIM P-1)的水平及其相关影响因素,探讨M M P-3及TIM P-1在R A的作用机制。方法选择41例初诊活动期RA患者和30名正常健康志愿者,以酶联免疫吸附试验(ELISA)分别检测血清M M P-3及TIM P-1水平,计算M M P-3/TIM P-1,同时测定关节功能、X线、关节肿胀数(SJC)、血沉(ESR)、类风湿因子(R F)、C反应蛋白(CR P)等相关实验室指标。结果活动期R A患者血清M M P-3、TIM P-1明显增高(P<0.01),且以M M P-3增高更为显著,M M P-3/TIM P-1较正常组亦增高(P<0.05)。不同关节功能分级时,上述指标差异无统计学意义(P>0.05),而不同X线分期时,各指标差异有统计学意义(P<0.01或P<0.05)。M M P-3、M M P-3/TIM P-1与SJC(P<0.01)、CRP(P<0.01)、ESR(P<0.05)呈正相关,二者与年龄、病程、晨僵时间、RF无明显相关性(P>0.05)。结论M M P-3、TIM P-1在RA血清中高水平存在,二者比例失衡导致RA发生。M M P-3、M M P-3/TIM P-1的高低可作为反映病情活动及预后的指标,阻断M M P-3高水平有可能成为治疗RA的新途径之一。展开更多
Objective： To explore the influence of charred Gossamer urocteae （CGU） on the functions of primary cultured mouse oral fibroblasts and reveal its mechanism in wound healing. Methods： CGU was extracted with differe...Objective： To explore the influence of charred Gossamer urocteae （CGU） on the functions of primary cultured mouse oral fibroblasts and reveal its mechanism in wound healing. Methods： CGU was extracted with different solvents and ethanol extract （EE）, ethyl acetate fraction （EF）, n-butanol fraction （BF） and aqueous fraction （AF） were obtained. The effects of different fractions on the proliferation, matrix metaUoproteinase-2,9 （MMP-2,9） activities, synthesis of collagen and tissue inhibitor of metalloproteinase 1 （TIMP-1） in the mouse oral fibroblasts were determined by MTT, gelatin zymography, chloramine-T method, and enzyme-linked immunosorbent assay （ELISA） respectively. Results： EE, EF and BF at high concentrations could significantly inhibit proliferation of fibroblasts （P〈0.05 or P〈0.01）, and at low concentrations EF and BF could promote proliferation of fibroblasts, and BF and AF could significantly inhibit collagen synthesis （P〈0.05 or P〈0.01）. EE, EF and AF at high concentrations could significantly increase the MMP-9 activity, and BF and AF could significantly inhibit synthesis of TIMP-1. Conclusion： CGU at high concentrations can inhibit the proliferations of fibroblasts and synthesis of collagen, and in healing of wound, CGU at high concentrations possibly has the functions of anti-fibrosis and anti-scar, and the mechanism to promote degradation of collagen is possibly related to the increase in MMP-9 activity and the inhibition of TIMP-1 synthesis.展开更多
文摘Objective To investigate the regulatory effect of multifactor on the matrix metalloproteinases-9 （MMP-9） and the tissue inhibitor of metalloproteinase-1 （TIMP-1） in endometrial stromal cells. Methods The endometrial stromal cells separated from the proliferative endometrial tissues were incubated with medium alone, 17-β estradiol （E2,10^-8 mol/L）, medroxyprogesterone acetate （MPA, 10^-6 mol/L）, E2（10^-8 mol/L）＋MPA （10^-6 mol/L）, E2 （10^-8 mol/L）＋MPA （10^-6 mol/L）＋RU486 （10^-5 mol/L） or HB-EGF （10 ng/ml） for 48 h respectively. The expressions of MMP-9 and TIMP-1 were detected by in situ hybridization, immunocytochemistry, reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blotting. Results Compared with control group [mRNA, 0. 729 ± 0. 090 （MMP-9） and 1.056± 0.154 （TIMP-1）; protein, 0.545 ±0.086 （MMP-9） and 0.745 ±0.154 （TIMP-1）], expressions of MMP-9 and TIMP-1 in E2 alone, progestin alone or E2 combined with progestin group were respectively：mRNA, 0.413 ± 0.069, 0.402 ± 0.073 and 0.407 ± 0.039; 0.487 ± 0.093, 0.503 ± 0.093 and 0.468 ± 0.075：protein, 0.294 ± 0.076, 0.331 ±0.064 and 0.265 ±0.049; 0.425 ±0.085, 0.397 ±0.065 and 0.435 ± 0.099. RU486 weakened the expression level of down-regulation, while HB-EGF elevated the level of MMP-9 and TIMP-1 after 48 h treatment （mRNA, 0.955 ± 0.068 and 1.396 ± 0.238; protein, 0. 780 ± 0.109 and 0.985 ± 0.165）. Conclusions 1） Both E2 and progestin can down-regulate the expressions of MMP-9 and TIMP-1 in endometrial stromal cells, but RU486 can inhibit the effect. 2） HB-EGF can elevate the level of MMP-9 and TIMP-1. 3） E2, progestin and HB-EGF have effect on the ratio of MM-P/TIMP-1.
文摘AIM： To evaluate the effects of angiopoietin-1 （Ang-1） on adhesion of gastric cancer cell line BGC-823 and expression of integrin β1, CD44V6, urokinase-type plasminogen activator （uPA） and matrix metalloproteinase-2 （MMP-2）. METHODS： BGC-823 cells were transfected transiently with adenovirus-Ang-1 （Ad-Ang-1）. Cells transfected transiently with adenovirus-green fluorescent protein （Ad-GFP） and untransfected cells were used as a negative and blank control group, respectively. The cell adhesion rate between cell and extracellular matrix （ECM） was determined by cell adhesion assay. To investigate whether Ang-1 could reinforce gastric carcinoma metastasis, we performed migration and invasion assays in BGC-823 cells. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 were detected by reverse transcription polymerase chain reaction and Western blotting, respectively. The expression of integrin β1 and CD44V6 was measured by immunohistochemistry. RESULTS： BGC-823 cells were transfected successfully. The adhesion rate increased significantly in the Ad-Ang-1 group （P 〈 0.05）. The Ad-Ang-1-transfected group had a significant increase in migration and invasion compared with that of the mock-transfected and Ad-GFP groups. The mRNA and protein expression of integrin β1, CD44V6, uPA and MMP-2 in the Ad- Ang-1 group was higher than that in the Ad-GFP and blank control groups （P 〈 0.05）. Compared with mocktransfected and Ad-GFP groups, integrin 131 and CD44V6 expression intensity greatly increased （P 〈 0.05）. CONCLUSION： Transfection of Ang-1 into human gastric cancer cell line BGC-823 can significantly increase expression of integrin β1 and CD44V6, by which cell adhesion and metastasis to the ECM are promoted.
基金supported by the project of Shanghai Municipal Health Bureau(2011160)
文摘Objective To investigate the role of matrix metalloproteinase-1 （MMP-1）/protease- activated receptor-1 （PAR-l） signaling in the cervical cancer invasion. Methods RT-PCR was used to test the mRNA level of MMP-1 in 59 cases of cervical squamous cell cancer. Cell invasion was evaluated by Transwell assay to explore the effect of adding human recombinant MMP-1 （rhMMP-1） and PARI-siRNA on cervical cancer invasion. Results In cervical cancer tissues, more MMP-1 expression was observed than that in the normal cervical tissues, and its expression correlated with cancer status. Human recombinant MMP-1 （rhMMP-1） could promote Hela cell invasion, and its number of invasive cell correlated with the concentration of rhMMP-1. Disrupting the expression of PAR-1 reduced the MMP-1 promoting-effect on Hela cell invasion, but had no effect on non-MMP-1 invasive action. Conclusion The MMP-1/PAR-1 signaling pathway is involved in cervical cancer invasion. Therefore, blocking PAR-1 may represent a new therapeutic option for metastatic cervical cancer.
文摘目的研究活动期类风湿关节炎(rheum atoid arthritis,RA)患者血清基质金属蛋白酶-3(m atrix m etalloproteinase3,M M P-3)、金属蛋白酶组织抑制剂-1(tissue inhibitors ofm etalloproteinase1,TIM P-1)的水平及其相关影响因素,探讨M M P-3及TIM P-1在R A的作用机制。方法选择41例初诊活动期RA患者和30名正常健康志愿者,以酶联免疫吸附试验(ELISA)分别检测血清M M P-3及TIM P-1水平,计算M M P-3/TIM P-1,同时测定关节功能、X线、关节肿胀数(SJC)、血沉(ESR)、类风湿因子(R F)、C反应蛋白(CR P)等相关实验室指标。结果活动期R A患者血清M M P-3、TIM P-1明显增高(P<0.01),且以M M P-3增高更为显著,M M P-3/TIM P-1较正常组亦增高(P<0.05)。不同关节功能分级时,上述指标差异无统计学意义(P>0.05),而不同X线分期时,各指标差异有统计学意义(P<0.01或P<0.05)。M M P-3、M M P-3/TIM P-1与SJC(P<0.01)、CRP(P<0.01)、ESR(P<0.05)呈正相关,二者与年龄、病程、晨僵时间、RF无明显相关性(P>0.05)。结论M M P-3、TIM P-1在RA血清中高水平存在,二者比例失衡导致RA发生。M M P-3、M M P-3/TIM P-1的高低可作为反映病情活动及预后的指标,阻断M M P-3高水平有可能成为治疗RA的新途径之一。
文摘Objective： To explore the influence of charred Gossamer urocteae （CGU） on the functions of primary cultured mouse oral fibroblasts and reveal its mechanism in wound healing. Methods： CGU was extracted with different solvents and ethanol extract （EE）, ethyl acetate fraction （EF）, n-butanol fraction （BF） and aqueous fraction （AF） were obtained. The effects of different fractions on the proliferation, matrix metaUoproteinase-2,9 （MMP-2,9） activities, synthesis of collagen and tissue inhibitor of metalloproteinase 1 （TIMP-1） in the mouse oral fibroblasts were determined by MTT, gelatin zymography, chloramine-T method, and enzyme-linked immunosorbent assay （ELISA） respectively. Results： EE, EF and BF at high concentrations could significantly inhibit proliferation of fibroblasts （P〈0.05 or P〈0.01）, and at low concentrations EF and BF could promote proliferation of fibroblasts, and BF and AF could significantly inhibit collagen synthesis （P〈0.05 or P〈0.01）. EE, EF and AF at high concentrations could significantly increase the MMP-9 activity, and BF and AF could significantly inhibit synthesis of TIMP-1. Conclusion： CGU at high concentrations can inhibit the proliferations of fibroblasts and synthesis of collagen, and in healing of wound, CGU at high concentrations possibly has the functions of anti-fibrosis and anti-scar, and the mechanism to promote degradation of collagen is possibly related to the increase in MMP-9 activity and the inhibition of TIMP-1 synthesis.
