PSD-007对人乳腺癌MCF-7细胞的光动力杀伤效应  

Study on the photodynamic effect of PSD-007 on human breast neoplasm cell line MCF-7

作　　者：张洪梅[1,2] 王宏[2] 陈洪丽[2] 李迎新[2] 徐应军[1] 关维俊[1] 顾立超 陈乃耀[3] 

机构地区：[1]河北联合大学公共卫生学院流行病与卫生统计学教研室,河北唐山063000 [2]中国医学科学院生物医学工程研究所,天津300192 [3]河北联合大学附属医院血液科,河北唐山063000

出　　处：《山东大学学报（医学版）》2014年第2期52-57,共6页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(31271434);河北省自然科学基金(H2013209253)

摘　　要：目的探讨癌光啉(PSD-007)对人乳腺癌MCF-7细胞体外光动力杀伤效应。方法噻唑蓝(MTT)比色法检测不同浓度PSD-007、不同能量光照MCF-7细胞的光密度(OD)值;以不同能量(1.2、2.4、4.8 J/cm2)激光照射(分别为3个实验组)10μg/mL PSD-007孵育的细胞,彗星实验、DAPI染色观察细胞形态学改变,流式细胞术(FCM)分析细胞周期及细胞凋亡率,免疫印记(Western blotting)检测B细胞淋巴瘤/白血病-2蛋白(Bcl-2)、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3蛋白(Caspase-3)表达量,同时设空白对照、单纯光照和单纯光敏剂组为对照组。结果光动力治疗对MCF-7细胞有明显的杀伤效应。彗星图像可见明显慧尾存在。DAPI染色观察到细胞呈现典型的凋亡形态特征。FCM分析结果显示,细胞凋亡率与空白对照组比较差异有统计学意义(P<0.05),凋亡率随光照能量增大而增高,G0/G1期细胞比例明显增加,细胞周期进程减慢。Western blotting结果显示,相比对照组,实验组Caspase-3和Bax表达水平明显上调,而Bcl-2表达水平下调。结论 PSD-007对人乳腺癌细胞系MCF-7细胞具有光动力杀伤效应,其杀伤效应与Bcl-2、Bax、Caspase-3密切相关。Objective To investigate the photodynamic effect of PSD-007 on human breast neoplasm cell line MCF-7 in vitro. Methods MTT assay was performed to examine MCF-7 cells treated with different concentrations of PSD-007 and different intensities of laser. Different intensities of laser （ 1.2, 2.4, 4.8 J/cm2 ） were applied to treat MCF-7 cells with 10μg/mL of PSD-007. The morphological changes of MCF-7cells were observed with comet assay and DAPI stai- ning. The cellular apoptosis rate and cell cycle phase distribution of MCF-7 cells were measured with flow cytometry （FCM）. The relative expressions of Bcl-2, Bax, and Caspase-3 proteins were detected with Western blotting. The blank control group, pure laser group and pure photosensitizer group were established as controls. Results Significant differences in the inhibitory effects were observed （ P 〈 0.05 ）. The comet figure significantly observed the comet tail. The morphological study found that multinucleate giant cells existed in the apoptotic cells in PSD-007 PDT. The FCM assay showed that the apoptosis rates were obvious and the killing effect improved with the intensity of laser （P 〈 0.05）. The results of cell cycle analysis revealed a significant increase of the number of cells in G0/G1 phase. Com- pared with the control groups, Caspase-3 and Bax expressions were markedly up-regulated in the experiment groups,while Bcl-2 expression was significantly down-regulated. Conclusion PSD-007 has a photodynamic effect on human breast carcinoma cell line MCF-7 in vitro, which is closely associated with Caspase-3, Bcl-2 and Bax.

关 键 词：光动力疗法 乳腺肿瘤 MCF-7细胞 癌光啉 

分 类 号：R318.51[医药卫生—生物医学工程]