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肺癌死亡率居世界肿瘤首位,5年生存率不足15%,顺铂耐药是死亡率居高不下的重要原因,本文将探讨光敏剂MPPa介导的光动力疗法诱导人非小细胞肺癌顺铂耐药株A549/DDP发生凋亡的现象及其机制。课题组分别给予不同浓度光敏剂MPPa(0、1、2、4、8、16μmol/L)、不同能量光照(0、0.6、1.2、2.4、3.6、4.8J/cm2)处理细胞,CCK-8法检测细胞活性。将细胞分为对照组、MPPa组(2μmol/L MPPa)、光照组(2.4J/cm2光能量密度)和MPPa介导光动力组(PDT组)(2μmol/L MPPa、2.4J/cm2光能量密度)。应用Anne-V/PI双染流式细胞技术检测细胞凋亡;DCFH-DA染色观察细胞内活性氧产生;蛋白质印迹法(Western blot)检测B淋巴细胞瘤-2(Bcl-2)蛋白和Bcl-2相关X蛋白(Bax)的表达。实验结果显示,单纯光照及MPPa对细胞生长无抑制作用;MPPa介导光动力却对人肺癌耐顺铂细胞A549/DDP有显著杀伤作用,其杀伤作用呈明显的剂量效应关系(P<0.05);PDT组发生凋亡率均高于各对照组(P<0.05);DCFH-DA染色发现PDT组细胞内活性氧明显高于各对照组;Western blot检测发现PDT组Bax蛋白表达升高,Bcl-2蛋白表达降低。实验证实MPPa介导的光动力疗法可抑制肺癌顺铂耐药细胞A549/DDP活性,并诱导其凋亡。
Lung cancer mortality rate ranks first in the world, the 5-year survival rate of less than 15%, cisplatin resistance is an important reason for the high mortality rate, this paper will explore the photosensitizer MPPa-mediated photodynamic therapy induced non-small cell lung cancer cisplatin The Phenotype and Mechanism of Apoptosis in A549 / DDP Cells. The groups were given different concentrations of photosensitizer MPPa (0,1,2,4,8,16μmol / L), different energy light (0,0.6,1.2,2.4,3.6,4.8 J / cm2) treatment of cells, CCK-8 Method to detect cell activity. The cells were divided into control group, MPPa group (2μmol / L MPPa), light group (2.4J / cm2 light energy density) and MPPa-mediated PDT group (2μmol / L MPPa, 2.4J / density). Apoptosis was detected by Anne-V / PI double staining flow cytometry; reactive oxygen species (ROS) production was observed by DCFH-DA staining; Bcl-2 protein and Bcl- -2 related X protein (Bax) expression. The experimental results showed that light and MPPa had no inhibitory effect on cell growth. MPPa-mediated photodynamic therapy had a significant killing effect on human lung cancer cell line A549 / DDP. The killing effect was dose-dependent (P <0.05) ; The apoptosis rate in PDT group was higher than that in each control group (P <0.05); DCFH-DA staining found that intracellular reactive oxygen species in PDT group was significantly higher than that in each control group; Western blot showed that Bax protein expression increased, -2 protein expression decreased. Experiments confirmed that MPPa-mediated photodynamic therapy can inhibit lung cancer cisplatin-resistant cell A549 / DDP activity and induce apoptosis.