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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 investigate the cytotoxic mechanism of singlet oxygen and oxide in Photofrin-Diomed 630-PDT. Methods Human immortalized esophageal epithelial cell line SHEE and its malignant cell line SHEEC were randomly divided into 20 groups. After 24 h inoculation, M199 complete medium was replaced with 100 μL of serum free 30 μg / mL Photofrin-Ⅱ solution and incubated After 150 minutes, M199 solution with different concentrations of sodium azide (NaN3) and superoxide dismutase (SOD) was rapidly replaced in the immediate intervention group and irradiated with light within 15 min. The M199 complete medium was replaced with light and the culture was continued 24 h. The M199 complete medium was no longer replaced by the continuous intervention group until 24 h, then the CCK-8 method was used to test the survival rate of the two cell lines. Results The immediate antagonism of Photofrin-Diomed 630-PDT induced by NaN3 on SHEE and SHEEC cell lines was enhanced with the increase of NaN3 concentration. The continuous intervention of NaN3 on SHEEC and SHEEC cells by Photofrin-Diomed 630-PDT showed cytotoxicity. There was no antagonism of SOD on Photofrin-Diomed 630-PDT in cell lines SHEE and SHEEC, and only cytotoxicity was observed. Conclusions The immediate intervention of NaN3 on Photofrin-Diomed 630-PDT in human immortalized esophageal epithelial cell line SHEE and SHEEC suggests that the mechanism of PDT cell damage may mainly be the instantaneous killing effect of singlet oxygen, but not obvious with other oxidants.