人永生化食管上皮细胞系SHEE及其癌变细胞系SHEEC的Photofrin-Diomed630-PDT损伤机制  

Mechanism of photodynamic damage on human immortalization esophageal epithelial cell line SHEE and the malignant transformation cell line SHEEC by Photofrin-Diomed 630-PDT

作　　者：高社干[1,2] 潘利咸[1] 王立东[2] 冯笑山[1,2] 曲智锋[1] 单探幽[1] 谢萱虎[1] 贾瑞诺[1] 宋娜莎[1] 

机构地区：[1]河南科技大学第一附属医院肿瘤科,河南科技大学肿瘤研究所,河南洛阳471003 [2]郑州大学基础医学院癌症研究室,河南省食管癌重点开放实验室,河南郑州450052

出　　处：《西安交通大学学报（医学版）》2011年第3期290-294,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.30670956);河南省2007年科技发展计划项目(No.072102310086)~~

摘　　要：目的探讨单态氧与氧化物在Photofrin-Diomed 630-PDT中的细胞毒作用机制。方法将人永生化食管上皮细胞系SHEE及其癌变细胞系SHEEC分别随机分成20组,接种24 h贴壁后,更换M199完全培养液为不含血清的30μg/mL的Photofrin-Ⅱ溶液100μL,孵育150 min后,即时干预组迅速更换不同浓度叠氮化钠(NaN3)和超氧化物歧化酶(SOD)的M199液,在15 min内进行光照处理,光照处理后更换M199完全培养液,继续培养24 h;持续干预组光照处理后不再更换M199完全培养液,直至24 h检测,然后应用CCK-8法检测两个细胞系各组的存活率。结果 NaN3对细胞系SHEE和SHEEC的Photofrin-Diomed 630-PDT即时干预随NaN3浓度的增加其拮抗作用也增强;NaN3对细胞系SHEE和SHEEC的Photofrin-Diomed 630-PDT持续干预表现为细胞毒副作用;SOD对细胞系SHEE和SHEEC的Photofrin-Diomed 630-PDT干预无拮抗作用,仅表现细胞毒作用。结论 NaN3对人永生化食管上皮细胞系SHEE和SHEEC的Photofrin-Diomed 630-PDT的即时干预,提示肿瘤PDT细胞损伤的机制可能主要为单态氧的瞬间杀伤作用,而与其他氧化剂关系不明显。Objective To explore the cytotoxic effect of singlet oxygen and oxidants on Photofrin-Diomed 630-PDT.Methods The human immortalization esophageal epithelial cell line SHEE and the malignant transformation cell line SHEEC were divided into 20 groups randomly.After 24-hour inoculation,the cells accreted on paries of culture capsule completely and we replaced M199 complete culture solution with 100 μL Photofrin-Ⅱ solution（30 μg/mL）.After 150 minutes’ incubation,we rapidly changed M199 with different concentrations of NaN3 and SOD solution in the instantaneous intervention group and treated it with PDT for 15 min,then removed the NaN3 and SOD solution,and replaced it with M199 complete culture solution.We examined the inhibitory effect on cell line SHEE and SHEEC under photodynamic therapy with CCK-8 method after 24-hour cultivation;but in the instantaneous intervention group,NaN3 and SOD solution was retained until the samples were examined after 24 h.Results The antagonism of NaN3 to Photofrin-Diomed 630-PDT on cell line SHEE and SHEEC increased with the increase of NaN3 concentration in the instantaneous intervention group.However,NaN3 showed a cytotoxic effect Supported by the National Natural Science Foundation of China（No.30670956） and the Sci-tech Research Development Project of Henan Province（No.072102310086）.on PDT with the increase of NaN3 concentration in the continuous intervention group.SOD showed a cytotoxic effect on cell line SHEE and SHEEC with Photofrin-Diomed 630-PDT,and no antagonism was observed.Conclusion The antagonism of NaN3 to Photofrin-Diomed 630-PDT on human immortalization esophageal epithelial cell line SHEE indicates that instantaneous lethal effect by singlet oxygen may play a very important role in the mechanism of PDT,but other oxidants may have little to do with PDT

关 键 词：细胞系 SHEE SHEEC 光动力治疗 Photofrin-Ⅱ 单态氧 叠氮化钠 超氧化物歧化酶 

分 类 号：R730.5[医药卫生—肿瘤]