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光敏剂MPPa介导光动力诱导人乳腺癌细胞MCF-7的内质网应激途径凋亡

, PP. 2115-2120

Keywords: 内质网应激,凋亡,光动力治疗,MPPa

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Abstract:

目的探讨光敏剂MPPa(pyropheophorbide-amethylester)介导的光动力作用于人乳腺癌细胞MCF-7,诱导内质网应激途径凋亡的机制。方法给予不同浓度光敏剂MPPa(0、0.5、1、2、4μmol/L),以及不同能量光照(0、0.9、1.8、2.7、3.6J/cm2)处理MCF-7细胞,用CCK-8法检测细胞活力。确定2μmol/LMPPa及1.8J/cm2光能量为实验剂量。激光共聚焦显微镜观察光敏剂MPPa(2μmol/L)内质网亚细胞器定位。将MCF-7细胞进行完全随机化分组,分为MPPa-PDT组(给予2μmol/LMPPa及1.8J/cm2光能量)、空白对照组、单纯MPPa组(2μmol/LMPPa)和单纯光照组(1.8J/cm2光能量)。应用AnnexinV-FITC/PI双染流式细胞仪检测细胞的凋亡率。Flou-3/MA流式细胞仪检测细胞内Ca2+浓度变化。Westernblot检测内质网应激标志蛋白BIP和内质网应激途径相关凋亡蛋白caspase-12、CHOP的表达。结果MPPa介导光动力对MCF-7细胞有显著杀伤作用,抑制其生长,且抑制作用呈明显的剂量效应关系(P<0.05);MPPa可富集在内质网上;MPPa-PDT组凋亡率显著高于各对照组(P<0.05);MPPa-PDT组细胞内Ca2+荧光强度较对照组增高(P<0.05);Westernblot检测发现BIP、caspase-12、CHOP蛋白在MPPa-PDT组中各时间点表达升高,而在各对照组中表达较低。结论MPPa介导的光动力可诱导MCF-7细胞发生内质网应激途径凋亡。

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