Angiotensin II (Ang II) is the main mediator of the Renin-Angiotensin-System acting on AT<sub>1</sub> and other AT receptors. It is regarded as a pleiotropic agent that induces many actions, including func...Angiotensin II (Ang II) is the main mediator of the Renin-Angiotensin-System acting on AT<sub>1</sub> and other AT receptors. It is regarded as a pleiotropic agent that induces many actions, including functioning as a growth factor, and as a contractile hormone, among others. The aim of this work was to examine the impact of Ang II on the expression and function of α<sub>1</sub>-adrenergic receptors (α<sub>1</sub>-ARs) in cultured rat aorta, and aorta-derived smooth muscle cells. Isolated Wistar rat aorta was incubated for 24 h in DMEM at 37˚C, then subjected to isometric tension and to the action of added norepinephrine, in concentration-response curves. Ang II was added (1 × 10<sup>−5</sup> M), and in some experiments, 5-Methylurapidil (α<sub>1A</sub>-AR antagonist), AH11110A (α<sub>1B</sub>-AR antagonist), or BMY-7378 (α<sub>1D</sub>-AR antagonist), were used to identify the α<sub>1</sub>-AR involved in the response. Desensitization of the contractile response to norepinephrine was observed due to incubation time, and by the Ang II action. α<sub>1D</sub>-AR was protected from desensitization by BMY-7378;while RS-100329 and prazosin partially mitigated desensitization. In another set of experiments, isolated aorta-derived smooth muscle cells were exposed to Ang II and α<sub>1</sub>-ARs proteins were evaluated. α<sub>1D</sub>-AR increased at 30 and 60 min post Ang II exposure, the α<sub>1A</sub>-AR diminished from 1 to 4 h, while α<sub>1B</sub>-AR remained unchanged over 24 h of Ang II exposure. Ang II induced an increase of α<sub>1D</sub>-AR at short times, and BMY-7378 protected α<sub>1D</sub>-AR from desensitization.展开更多
目的探讨癌症和帕金森病相关蛋白DJ-1(parkinson disease protein 7,oncogene DJ-1,DJ-1)下调磷酸酯酶与张力蛋白同源物(phosphatase and tensin homolog,PTEN)诱导雄激素受体(androgen receptor,AR)表达促进非激素依赖性乳腺癌转移的...目的探讨癌症和帕金森病相关蛋白DJ-1(parkinson disease protein 7,oncogene DJ-1,DJ-1)下调磷酸酯酶与张力蛋白同源物(phosphatase and tensin homolog,PTEN)诱导雄激素受体(androgen receptor,AR)表达促进非激素依赖性乳腺癌转移的机制。方法构建人绿色荧光蛋白表达载体pEGFP/DJ-1;Western blotting对比分析过表达DJ-1基因后MDA-MB-231细胞DJ-1、PTEN、雄激素受体(androgen receptor,AR)的表达;体外迁移与侵袭实验检测过表达DJ-1基因后对MDA-MB-231细胞的迁移与侵袭的影响。收集转移的非激素依赖性乳腺癌和未出现转移的非激素依赖性乳腺癌手术切除标本各40例,采用免疫组织化学检测DJ-1蛋白的表达。结果 Western blotting发现经过转染人DJ-1基因的绿色荧光蛋白表达载体pEGFP/DJ-1后,MDA-MB-231细胞DJ-1的表达水平升高,PTEN的表达水平下降,AR水平升高。体外迁移与侵袭实验发现,转染pEGFP/DJ-1促进SWO-38细胞的迁移和侵袭,与转染空白对照组比较,差异有统计学意义(P<0.05)。发生转移非激素依赖性乳腺癌组织中DJ-1阳性表达率(85.0%)明显高于未发生转移乳腺癌组织(55.0%)(P<0.05)。结论 DJ-1能促进非激素依赖性乳腺癌的迁移与侵袭。这可能与DJ-1抑制PTEN表达,提高AR水平从而促进肿瘤细胞的迁移侵袭能力有关。展开更多
文摘Angiotensin II (Ang II) is the main mediator of the Renin-Angiotensin-System acting on AT<sub>1</sub> and other AT receptors. It is regarded as a pleiotropic agent that induces many actions, including functioning as a growth factor, and as a contractile hormone, among others. The aim of this work was to examine the impact of Ang II on the expression and function of α<sub>1</sub>-adrenergic receptors (α<sub>1</sub>-ARs) in cultured rat aorta, and aorta-derived smooth muscle cells. Isolated Wistar rat aorta was incubated for 24 h in DMEM at 37˚C, then subjected to isometric tension and to the action of added norepinephrine, in concentration-response curves. Ang II was added (1 × 10<sup>−5</sup> M), and in some experiments, 5-Methylurapidil (α<sub>1A</sub>-AR antagonist), AH11110A (α<sub>1B</sub>-AR antagonist), or BMY-7378 (α<sub>1D</sub>-AR antagonist), were used to identify the α<sub>1</sub>-AR involved in the response. Desensitization of the contractile response to norepinephrine was observed due to incubation time, and by the Ang II action. α<sub>1D</sub>-AR was protected from desensitization by BMY-7378;while RS-100329 and prazosin partially mitigated desensitization. In another set of experiments, isolated aorta-derived smooth muscle cells were exposed to Ang II and α<sub>1</sub>-ARs proteins were evaluated. α<sub>1D</sub>-AR increased at 30 and 60 min post Ang II exposure, the α<sub>1A</sub>-AR diminished from 1 to 4 h, while α<sub>1B</sub>-AR remained unchanged over 24 h of Ang II exposure. Ang II induced an increase of α<sub>1D</sub>-AR at short times, and BMY-7378 protected α<sub>1D</sub>-AR from desensitization.
文摘目的探讨癌症和帕金森病相关蛋白DJ-1(parkinson disease protein 7,oncogene DJ-1,DJ-1)下调磷酸酯酶与张力蛋白同源物(phosphatase and tensin homolog,PTEN)诱导雄激素受体(androgen receptor,AR)表达促进非激素依赖性乳腺癌转移的机制。方法构建人绿色荧光蛋白表达载体pEGFP/DJ-1;Western blotting对比分析过表达DJ-1基因后MDA-MB-231细胞DJ-1、PTEN、雄激素受体(androgen receptor,AR)的表达;体外迁移与侵袭实验检测过表达DJ-1基因后对MDA-MB-231细胞的迁移与侵袭的影响。收集转移的非激素依赖性乳腺癌和未出现转移的非激素依赖性乳腺癌手术切除标本各40例,采用免疫组织化学检测DJ-1蛋白的表达。结果 Western blotting发现经过转染人DJ-1基因的绿色荧光蛋白表达载体pEGFP/DJ-1后,MDA-MB-231细胞DJ-1的表达水平升高,PTEN的表达水平下降,AR水平升高。体外迁移与侵袭实验发现,转染pEGFP/DJ-1促进SWO-38细胞的迁移和侵袭,与转染空白对照组比较,差异有统计学意义(P<0.05)。发生转移非激素依赖性乳腺癌组织中DJ-1阳性表达率(85.0%)明显高于未发生转移乳腺癌组织(55.0%)(P<0.05)。结论 DJ-1能促进非激素依赖性乳腺癌的迁移与侵袭。这可能与DJ-1抑制PTEN表达,提高AR水平从而促进肿瘤细胞的迁移侵袭能力有关。
