激光照射下硫化铜纳米颗粒对Hela细胞的杀伤作用  

KILLING EFFECT OF COPPER SULFIDE NANOPARTICLES ON HELA CELLS UNDER LASER RADIATION

作　　者：葛南 刘树亮 高宏志[2] 张亚萍 王礼兴[2] 崔莲花[1] GE Nan;LIU Shuliang;GAO Hongzhi;ZHANG Yaping;WANG Lixing;CUI Lianhua(School of Public Health, Qingdao University, Qingdao 2GG021, China)

机构地区：[1]青岛大学公共卫生学院,山东青岛266021 [2]福建医科大学附属第二医院中心实验室

出　　处：《青岛大学学报（医学版）》2018年第1期17-22,共6页Journal of Qingdao University(Medical Sciences)

基　　金：山东省自然科学基金资助项目(ZR2011HM031)

摘　　要：目的探讨新型光热剂硫化铜纳米颗粒(BSA-CuS NPs)对HeLa细胞的光热杀伤作用及机制。方法以牛血清清蛋白(BSA)为生物模板合成BSA-CuS NPs,对其进行物理表征和光热转换实验。将HeLa细胞分为对照组、硫化铜组、光照组和联合组。采用CCK-8法检测BSA-CuS NPs的细胞毒性,TUNEL实验检测细胞的凋亡情况,酶标法检测各组细胞内超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)的活力,免疫蛋白印迹分析细胞色素C(Cyt-c)蛋白的表达。结果 BSA-CuS NPs为直径(26.25±1.86)nm的片状结构,紫外线可见吸收峰位于980nm。升温曲线说明BSA-CuS NPs较去离子水溶液升温效果更加明显,呈现光照强度依赖性。CCK-8实验结果表明,在100mg/L浓度范围内培养24h,BSA-CuS NPs对HeLa细胞的生长无明显抑制。TUNEL实验结果表明,联合组细胞的凋亡情况较其余3组明显增强。氧化应激指标测定显示,硫化铜组、光照组的SOD、GSH-Px较对照组降低,差异有统计学意义(F=223.36、920.00,P<0.01);联合组与硫化铜组比较、联合组与光照组比较,SOD、GSH-Px均降低,差异有统计学意义(t=5.28~33.84,P<0.01)。免疫蛋白印迹分析结果表明,联合组的Cytc蛋白表达量较硫化铜组、光照组和对照组增加,差异有统计学意义(F=19.85,P<0.01)。结论 BSA-CuS NPs对HeLa细胞具有光热杀伤效应,机制可能与BSA-CuS NPs诱导大量自由基产生,进而启动凋亡基因程序有关。Objective To investigate the photothermal killing effect of copper sulfide nanoparticles（BSA-CuS NPs）,a new type of photothermal agent,on HeLa cells and its mechanism. Methods BSA-CuS NPs were synthesized using bovine serum albumin（BSA）as the biological template,and physical characterization and photothermal conversion experiment were performed.HeLa cells were divided into control group,copper sulfide group,radiation group,and combination group.CCK-8 assay was used to measure the cytotoxicity of BSA-CuS NPs;terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling（TUNEL）was used to measure cell apoptosis;ELISA was used to measure the activity of superoxide dismutase（SOD）and glutathione peroxidase（GSH-Px）;Western blotting was used to measure the expression of cytochrome C. Results BSA-CuS NPs had a patchy structure with a diameter of（26.25±1.86）nm,and the ultraviolet visible absorption peak was around 980 nm.The heating curves illustrated that the temperature of BSA-CuS NPs rose faster than that of deionized water solution,which was dependent on laser intensity.The results of the CCK-8 assay showed that BSA-CuS NPs had no marked inhibitory effect on the growth of HeLa cells after culture for 24 hat a concentration of 100 mg/L.The results of TUNEL showed that the combination group had significantly greater apoptosis than the other three groups.According to the measurement of oxidative stress markers,the copper sulfide group and the radiation group had significantly lower SOD and GSH-Px than the control group（F=223.36 and920.00,P〈0.01）;the combination group had significantly lower SOD and GSH-Px than the copper sulfide group and the radiation group（t=5.28-33.84,P〈0.01）.Western blotting showed that the combination group had significantly higher protein expression of cytochrome C than the other three groups（F=19.85,P〈0.01）. Conclusion BSA-CuS NPs exert a strong photothermal killing effect on HeLa cells,possibly by inducing the production of a large number of free

关 键 词：硫化铜 纳米颗粒 激光疗法 细胞凋亡 超氧化物类 

分 类 号：R730.5[医药卫生—肿瘤] R616.6[医药卫生—临床医学]