PDGF B-chain, a component of platelet-derived growth factor (PDGF), is encoded by PDGF B-chain gene. Recently, the transcriptional regulation of PDGF B-chain gene has been studied extensively. Some cisacting elements ...PDGF B-chain, a component of platelet-derived growth factor (PDGF), is encoded by PDGF B-chain gene. Recently, the transcriptional regulation of PDGF B-chain gene has been studied extensively. Some cisacting elements in the gene and some relevant trans-acting factors have been identified. Some research data demonstrated that the transcriptional regulation of PDGF-B chain gene was uncommon during some common cardiovascular diseases. The molecular mechanisms underlying the pathogenic factors-mediated transcription of the gene are begining to be understood, which will open a good respect for prevention and treatment of the common cardiovascular diseases.展开更多
目的:构建甲状腺癌基因-1(TC-1)真核表达质粒,观察TC-1癌蛋白高表达对人乳腺癌细胞迁移的影响。方法:用蛋白质印迹方法检测TC-1癌蛋白在培养的人MCF-7和BT549乳腺癌细胞中表达;从MCF-7细胞中提取总RNA,通过实时PCR(RT-PCR)扩增人TC-1 c ...目的:构建甲状腺癌基因-1(TC-1)真核表达质粒,观察TC-1癌蛋白高表达对人乳腺癌细胞迁移的影响。方法:用蛋白质印迹方法检测TC-1癌蛋白在培养的人MCF-7和BT549乳腺癌细胞中表达;从MCF-7细胞中提取总RNA,通过实时PCR(RT-PCR)扩增人TC-1 c DNA全长开放读码框架(ORF),并插入真核表达载体Flag-pc DNA3.0中构建Flag-TC-1-pc DNA3.0重组质粒;经限制性内切酶和DNA测序鉴定后,该重组质粒用于转染BT549细胞,进行蛋白质印迹分析鉴定Flag-TC-1融合蛋白表达;进行细胞划痕实验,观察TC-1对BT549细胞迁移的影响。结果:TC-1蛋白在人MCF-7乳腺癌细胞中显著表达,而BT549细胞中表达极低;人MCF-7乳腺癌细胞TC-1全长ORF的RT-PCR产物为337 bp;重组质粒Flag-TC-1-pc DNA3.0经Bam HⅠ和XhoⅠ双酶切出现特征性的327 bp片段;DNA测序证实TC-1全长ORF序列正确无误,以正确的阅读框架插入Flag-pc DNA3.0载体中。蛋白质印迹结果显示该重组质粒转染的BT549细胞表达的FlagTC-1融合蛋白可被抗Flag抗体特异性识别,分子质量约为13.6 k Da,符合预期;划痕实验结果显示,高表达TC-1的BT549细胞迁移性增加。结论:内源性TC-1蛋白在人BT549乳腺癌细胞中表达极低,Flag-TC-1融合蛋白高表达促进BT549细胞迁移。展开更多
Effect of endotoxin, and endotoxin plus Radix Salviae Miltiorrhizae （RSM） on bovine aortic endothelial cells （BAECs） for the production of tissue factor （TF）, plasminogen activator （PA） and plasminogen activat...Effect of endotoxin, and endotoxin plus Radix Salviae Miltiorrhizae （RSM） on bovine aortic endothelial cells （BAECs） for the production of tissue factor （TF）, plasminogen activator （PA） and plasminogen activator inhibitor （PAI） activities as well as prostacyclin （PGI<sub>2</sub>） content were studied. Stimulation of BAECs with endotoxin （1 μg/ ml） increased the expression of cell surface TF activity, the secretions of PAI activity and PGI<sub>2</sub> content. However, PA activity in endotoxin-treated groups didn’t change significantly in comparison with that of the control. RSM could stimulate the secretion of PA activity and prevent the increase in TF and PAI activities from endotoxin-treated BAECs. However, there was no significant difference in PGI<sub>2</sub> production between endotoxin-and endotoxin+RSM-treated BAECs. These results indicated that increased procoagulant function and decreased fibrinolytic activity of endothelial cells （ECs） might be one of the mechanisms of DIC in gram-negative septicemia and RSM would be a very important thug in the prevention and the treatment of DIC in gram-negative sepsis.展开更多
文摘PDGF B-chain, a component of platelet-derived growth factor (PDGF), is encoded by PDGF B-chain gene. Recently, the transcriptional regulation of PDGF B-chain gene has been studied extensively. Some cisacting elements in the gene and some relevant trans-acting factors have been identified. Some research data demonstrated that the transcriptional regulation of PDGF-B chain gene was uncommon during some common cardiovascular diseases. The molecular mechanisms underlying the pathogenic factors-mediated transcription of the gene are begining to be understood, which will open a good respect for prevention and treatment of the common cardiovascular diseases.
文摘Effect of endotoxin, and endotoxin plus Radix Salviae Miltiorrhizae （RSM） on bovine aortic endothelial cells （BAECs） for the production of tissue factor （TF）, plasminogen activator （PA） and plasminogen activator inhibitor （PAI） activities as well as prostacyclin （PGI<sub>2</sub>） content were studied. Stimulation of BAECs with endotoxin （1 μg/ ml） increased the expression of cell surface TF activity, the secretions of PAI activity and PGI<sub>2</sub> content. However, PA activity in endotoxin-treated groups didn’t change significantly in comparison with that of the control. RSM could stimulate the secretion of PA activity and prevent the increase in TF and PAI activities from endotoxin-treated BAECs. However, there was no significant difference in PGI<sub>2</sub> production between endotoxin-and endotoxin+RSM-treated BAECs. These results indicated that increased procoagulant function and decreased fibrinolytic activity of endothelial cells （ECs） might be one of the mechanisms of DIC in gram-negative septicemia and RSM would be a very important thug in the prevention and the treatment of DIC in gram-negative sepsis.
