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首页> 外文期刊>Radiation and Environmental Biophysics >Overexpression of truncated AIF regulated by Egr1 promoter radiation-induced apoptosis on MCF-7 cells
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Overexpression of truncated AIF regulated by Egr1 promoter radiation-induced apoptosis on MCF-7 cells

机译:Egr1启动子辐射诱导的MCF-7细胞凋亡调控的截短AIF的过表达

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摘要

It has been demonstrated that gene-radiotherapy can improve the radiotherapy by selectively increasing cells' response to ionizing radiation. Apoptosis-inducing factor (AIF) is a mitochondrial flavoprotein, and its C-terminal domain is responsible for the proapoptotic activity. In the present study, we overexpressed truncated AIF on MCF-7 cells by transfection of pcDNA3.1-tAIF (pc-tAIF) and pcDNA3.1-Egr1-tAIF (pc-Egr1-tAIF) plasmids. After MCF-7-tAIF cells were exposed to X-rays, the AIF and tAIF expressions, cell proliferation, apoptosis, cell cycle invasion, cytochrome c (Cyt c) release and activation of caspase-9 were measured by using Western blot, MTT assay, flow cytometry and Matrigel transwell assay, respectively. Our results showed that tAIF expression increased on time- and dose-dependent manners. Both tAIF and radiation can synergistically enhance the apoptosis, cell proliferation inhibition, cell cycle arrest and cell-invasive inhibition. In addition, tAIF overexpression and irradiation increased Cyt c release. However, only irradiation increased caspase-9 activation. Our studies indicated that tAIF overexpression might enhance apoptosis induced by radiation in MCF-7 cells.
机译:已经证明,基因放射疗法可以通过选择性地增加细胞对电离辐射的反应来改善放射疗法。凋亡诱导因子(AIF)是线粒体黄素蛋白,其C末端结构域负责促凋亡活性。在本研究中,我们通过转染pcDNA3.1-tAIF(pc-tAIF)和pcDNA3.1-Egr1-tAIF(pc-Egr1-tAIF)质粒在MCF-7细胞上过表达了截短的AIF。 X射线照射MCF-7-tAIF细胞后,采用Western blot,MTT法检测AIF和tAIF的表达,细胞增殖,凋亡,细胞周期侵袭,细胞色素c(Cyt c)的释放和胱天蛋白酶9的活化。检测,流式细胞术和Matrigel Transwell检测。我们的结果表明,tAIF表达以时间和剂量依赖性方式增加。 tAIF和辐射均可协同增强细胞凋亡,细胞增殖抑制,细胞周期停滞和细胞侵袭抑制。另外,tAIF过表达和辐射增加了Cyt c的释放。但是,仅辐射会增加caspase-9的激活。我们的研究表明,tAIF的过度表达可能增强MCF-7细胞辐射诱导的凋亡。

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