Background In a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explor...Background In a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explore the effect of CXCL12-CXCR4 axis on the metastasis of human ovarian cancer. Methods The expressions of CXCR4 and CXCL12 mRNA and protein in human ovarian cancer cell line CAOV-3 was detected by RT-PCR and immunocytochemistry. Methythiazolyltetraz.olium （MTT） was used to analyze the effect of different concentrations of CXCL12 on the proliferation of CAOV-3 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of CXCL12 and ascites on the migration and invasion of CAOV-3 cells. The expressions of integrin β1 and vascular endothelial growth factor-C （VEGF-C） mRNA were detected by RT-PCR. Data were analyzed using ANOVA by SAS 6.12. Results Under serum-free suboptimal culture conditions, CXCL12 （100 ng/ml） significantly enhanced the proliferation of CAOV-3 cells compared with the control and 10 ng/ml CXCL12 groups （0.428 ± 0.051 vs. 0.325 ±0.045 and 0.328±0.039, P〈0.05）. This enhancing effect of CXCL12 was significantly inhibited by 10 μg/ml neutralizing CXCR4 antibody or 1 μg/ml CXCR4 antagonist AMD3100. However, 10 μg/ml neutralizing CXCR4 antibody could not inhibit cell proliferation without CXCL12. The levels of migration and invasion of the CAOV-3 cells treated with 100 ng/ml CXCL12 were significantly higher than those in the control （migration： 523.3± 25.2 vs 108.0 ± 7.2; invasion： 39.3 ± 4.0 vs. 4.0 ± 1.0）. The enhancing effect of CXCL12 on cell migration and invasion increased with the concentration of CXCL12 （100 ng/ml vs 10 ng/ml： migration, 523.3 ± 25.2 vs 211.7 ± 24.7; invasion, 39.3 ± 4.0 vs 15.7 ± 3.1, P〈0.05）, and was strongly inhibited by 10 μg/ml neutralizing CXCR4 antibody or 1 μg/ml AMD3100. The number of migrated and invading cells in the CAOV-3 added with ascites was significantly higher than those in the 100 ng/ml CXCL12 group （migration： 706.6 ± 30.6 vs 523.3 ± 25.2, invasion： 61.7 ± 7.6 vs 39.3 ± 4.0, P〈0.05）. The level of integrin β1 mRNA was greatly increased at 3 hours after being treated with CXCL12 （0.53±0.10 vs. 1.53±0.16, P〈0.05）, and VEGF-C mRNA displayed significant augment at 24 hours after being treated with CXCL12 （0.52 ± 0.09 vs 1.11 ± 0.15, P〈0.05）. Conclusions CXCL12 and its receptor CXCR4 can promote the proliferation, migration, invasion of ovarian cancer cell line CAOV-3 and enhance its secretion of integrin β1 and VEGF-C. These effects can be inhibited by neutralizing CXCR4 antibody or AMD3100. CXCL12-CXCR4 axis plays an important role in ovarian cancer growth and metastasis.展开更多
Background Under an insulin resistance （IR） state, overproduction of reactive oxygen species （ROS） may be playing a major role in the pathogenesis of endothelial dysfunction, hypertension and atherosclerosis. Rece...Background Under an insulin resistance （IR） state, overproduction of reactive oxygen species （ROS） may be playing a major role in the pathogenesis of endothelial dysfunction, hypertension and atherosclerosis. Recently, increasing attention has been drawn to the beneficial effects of heme oxygenase-1 （HO-1） in the cardiovascular system. This study aimed to investigate the effects of HO-1 on vascular function of thoracic aorta in IR rats and demonstrate the probable mechanisms of HO-1 against endothelial dysfunction in IR states. Methods Sprague-Dawley （SD） rats fed with high-fat diet for 6 weeks and the IR models were validated with hyperinsulinemic-euglycemic clamp test. Then the IR rat models （n=-44） were further randomized into 3 subgroups, namely, the IR control group （n=26, in which 12 were sacrificed immediately and evaluated for all study measures）, a hemin treated IR group （n=-10） and a zinc protoporphyrin-Ⅸ （ZnPP-Ⅸ） treated IR group （n=-8） that were fed with a high-fat diet. Rats with standardized chow diet were used as the normal control group （n=-12）. The rats in IR control group, hemin treated IR group and ZnPP-Ⅸ treated IR group were subsequently treated every other day with an intraperitoneal injection of normal saline, hemin （inducer of HO-1, 30 μmol/kg） or ZnPP-Ⅸ （inhibitor of HO-1, 10 μmol/kg） for 4 weeks. Rats in the normal control group remained on a standardized chow diet and were treated with intraperitoneal injections of normal saline every other day for 4 weeks. Systolic arterial blood pressure （SABP） was measured by tail-cuffed microphotoelectric plethysmography. The blood carbon monoxide （CO） was measured by blood gas analysis. The levels of nitdc oxide （NO）, inducible nitric oxide synthase （iNOS）, endothelial nitric oxide synthase （eNOS）, blood glucose （BG）, insulin, total cholesterol （TC） and triglyceride （TG） in serum, and the levels of total antioxidant capacity （TAOC）, malondialdehyde （MDA） and superoxide dismutase （SOD） in the aorta were measured. The expression of HO-1 mRNA and HO-1 protein in aortal tissue were detected by semi-quantitative RT-PCR and Western blot. The vasoreacUve tensometry was performed with thoracic aortic rings （TARs）. Results Compared with the normal control group, the levels of SABP, BG, insulin, TC, TG, NO, iNOS and MDA were higher, while the levels of CO, TAOC, SOD and eNOS were lower in IR control rats. After treatment of IR rats for 4 weeks a more intensive expression of HO-1 mRNA and HO-1 protein were observed in hemin treated IR group compared with the normal control group. And compared with 4-week IR control rats, the levels of CO, TAOC, SOD and eNOS were increased, while the levels of SABP and iNOS activity were lower in the hemin treated IR group. Administration of hemin in IR rats appeared to improve the disordered vasorelaxation of TARs to acetylcholine （ACh）. Alternatively, the reverse results of SABP, CO, TAOC, SOD, iNOS and vasorelaxation responses to ACh were observed in IR rats with administration of ZnPP-Ⅸ. Conclusions The endothelial dysfunction in the aorta is present in the IR state. The protective effects of HO-1 against aortic endothelial dysfunction may be due to its antioxidation and regulative effect of vasoactive substances. It is proposed that hemin, inducer of HO-1, could be a potential therapeutic option for vascular dysfunction in IR states.展开更多
文摘Background In a previous study, we have verified that CXCR4 expression is correlated with tumor aggressive progression and poor prognosis in patients with epithelial ovarian cancer. The aim of this study was to explore the effect of CXCL12-CXCR4 axis on the metastasis of human ovarian cancer. Methods The expressions of CXCR4 and CXCL12 mRNA and protein in human ovarian cancer cell line CAOV-3 was detected by RT-PCR and immunocytochemistry. Methythiazolyltetraz.olium （MTT） was used to analyze the effect of different concentrations of CXCL12 on the proliferation of CAOV-3 cells. Transwell invasion chamber and matrigel were used to evaluate the effect of various concentrations of CXCL12 and ascites on the migration and invasion of CAOV-3 cells. The expressions of integrin β1 and vascular endothelial growth factor-C （VEGF-C） mRNA were detected by RT-PCR. Data were analyzed using ANOVA by SAS 6.12. Results Under serum-free suboptimal culture conditions, CXCL12 （100 ng/ml） significantly enhanced the proliferation of CAOV-3 cells compared with the control and 10 ng/ml CXCL12 groups （0.428 ± 0.051 vs. 0.325 ±0.045 and 0.328±0.039, P〈0.05）. This enhancing effect of CXCL12 was significantly inhibited by 10 μg/ml neutralizing CXCR4 antibody or 1 μg/ml CXCR4 antagonist AMD3100. However, 10 μg/ml neutralizing CXCR4 antibody could not inhibit cell proliferation without CXCL12. The levels of migration and invasion of the CAOV-3 cells treated with 100 ng/ml CXCL12 were significantly higher than those in the control （migration： 523.3± 25.2 vs 108.0 ± 7.2; invasion： 39.3 ± 4.0 vs. 4.0 ± 1.0）. The enhancing effect of CXCL12 on cell migration and invasion increased with the concentration of CXCL12 （100 ng/ml vs 10 ng/ml： migration, 523.3 ± 25.2 vs 211.7 ± 24.7; invasion, 39.3 ± 4.0 vs 15.7 ± 3.1, P〈0.05）, and was strongly inhibited by 10 μg/ml neutralizing CXCR4 antibody or 1 μg/ml AMD3100. The number of migrated and invading cells in the CAOV-3 added with ascites was significantly higher than those in the 100 ng/ml CXCL12 group （migration： 706.6 ± 30.6 vs 523.3 ± 25.2, invasion： 61.7 ± 7.6 vs 39.3 ± 4.0, P〈0.05）. The level of integrin β1 mRNA was greatly increased at 3 hours after being treated with CXCL12 （0.53±0.10 vs. 1.53±0.16, P〈0.05）, and VEGF-C mRNA displayed significant augment at 24 hours after being treated with CXCL12 （0.52 ± 0.09 vs 1.11 ± 0.15, P〈0.05）. Conclusions CXCL12 and its receptor CXCR4 can promote the proliferation, migration, invasion of ovarian cancer cell line CAOV-3 and enhance its secretion of integrin β1 and VEGF-C. These effects can be inhibited by neutralizing CXCR4 antibody or AMD3100. CXCL12-CXCR4 axis plays an important role in ovarian cancer growth and metastasis.
文摘Background Under an insulin resistance （IR） state, overproduction of reactive oxygen species （ROS） may be playing a major role in the pathogenesis of endothelial dysfunction, hypertension and atherosclerosis. Recently, increasing attention has been drawn to the beneficial effects of heme oxygenase-1 （HO-1） in the cardiovascular system. This study aimed to investigate the effects of HO-1 on vascular function of thoracic aorta in IR rats and demonstrate the probable mechanisms of HO-1 against endothelial dysfunction in IR states. Methods Sprague-Dawley （SD） rats fed with high-fat diet for 6 weeks and the IR models were validated with hyperinsulinemic-euglycemic clamp test. Then the IR rat models （n=-44） were further randomized into 3 subgroups, namely, the IR control group （n=26, in which 12 were sacrificed immediately and evaluated for all study measures）, a hemin treated IR group （n=-10） and a zinc protoporphyrin-Ⅸ （ZnPP-Ⅸ） treated IR group （n=-8） that were fed with a high-fat diet. Rats with standardized chow diet were used as the normal control group （n=-12）. The rats in IR control group, hemin treated IR group and ZnPP-Ⅸ treated IR group were subsequently treated every other day with an intraperitoneal injection of normal saline, hemin （inducer of HO-1, 30 μmol/kg） or ZnPP-Ⅸ （inhibitor of HO-1, 10 μmol/kg） for 4 weeks. Rats in the normal control group remained on a standardized chow diet and were treated with intraperitoneal injections of normal saline every other day for 4 weeks. Systolic arterial blood pressure （SABP） was measured by tail-cuffed microphotoelectric plethysmography. The blood carbon monoxide （CO） was measured by blood gas analysis. The levels of nitdc oxide （NO）, inducible nitric oxide synthase （iNOS）, endothelial nitric oxide synthase （eNOS）, blood glucose （BG）, insulin, total cholesterol （TC） and triglyceride （TG） in serum, and the levels of total antioxidant capacity （TAOC）, malondialdehyde （MDA） and superoxide dismutase （SOD） in the aorta were measured. The expression of HO-1 mRNA and HO-1 protein in aortal tissue were detected by semi-quantitative RT-PCR and Western blot. The vasoreacUve tensometry was performed with thoracic aortic rings （TARs）. Results Compared with the normal control group, the levels of SABP, BG, insulin, TC, TG, NO, iNOS and MDA were higher, while the levels of CO, TAOC, SOD and eNOS were lower in IR control rats. After treatment of IR rats for 4 weeks a more intensive expression of HO-1 mRNA and HO-1 protein were observed in hemin treated IR group compared with the normal control group. And compared with 4-week IR control rats, the levels of CO, TAOC, SOD and eNOS were increased, while the levels of SABP and iNOS activity were lower in the hemin treated IR group. Administration of hemin in IR rats appeared to improve the disordered vasorelaxation of TARs to acetylcholine （ACh）. Alternatively, the reverse results of SABP, CO, TAOC, SOD, iNOS and vasorelaxation responses to ACh were observed in IR rats with administration of ZnPP-Ⅸ. Conclusions The endothelial dysfunction in the aorta is present in the IR state. The protective effects of HO-1 against aortic endothelial dysfunction may be due to its antioxidation and regulative effect of vasoactive substances. It is proposed that hemin, inducer of HO-1, could be a potential therapeutic option for vascular dysfunction in IR states.
