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ANTP-SmacN7融合肽对H460细胞的辐射增敏作用
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ANTP-SmacN7融合肽对H460细胞的辐射增敏作用
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Journal Article
ANTP-SmacN7融合肽对H460细胞的辐射增敏作用
2016
Overview
背景与目的肿瘤的辐射耐受制约了放疗疗效,第二个线粒体衍生的半胱氨酸蛋白酶激活剂（Second mitochondria-derived activator of caspase,Smac）蛋白类似物可明显提高辐射诱导的肿瘤细胞凋亡,有望成为新型肿瘤辐射增敏药物。本研究旨在探讨新型Smac蛋白类似物ANTP-SmacN7融合肽对肺癌细胞系H460的辐射增敏作用。方法合成ANTP-SmacN7融合肽,连接荧光素FITC以观察融合肽能否进入细胞。对数生长期H460细胞分为空白对照组、单纯照射组、ANTP-SmacN7组和照射联合ANTP-SmacN7组,单纯照射组给予0 Gy、2 Gy、4 Gy、6 Gy照射,照射联合ANTP-SmacN7组中ANTP-SmacN7的浓度为20μmol/L,WST-1测定H460细胞的增殖。流式细胞仪测定细胞处理后24h和48 h的细胞凋亡率。Western blot实验检测caspase3和cleaved caspase3的表达水平。结果 ANTP-SmacN7融合能够顺利进入细胞,且能够增强H460细胞的辐射敏感性（F=25.1,P〈0.01,增敏比为1.86）,照射联合ANTP-SmacN7可明显降低H460细胞的克隆形成率（χ^2=45.2,P〈0.01;χ^2=40.3,P〈0.01）,提高cleaved caspase3的表达量,促进caspase3的活化,增加辐射诱导的细胞凋亡率。结论 ANTP-SmacN7融合肽可明显提高H460细胞的辐射敏感性,作为一种新的Smac蛋白类似物有望用于肿瘤的辐射增敏治疗。
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