CAJ | 학술논문

目的探讨一种新型光敏药物对人胃癌细胞MGC803的光动力学治疗(PDT)作用及其机制.方法采用bleaching光漂白法评价中国医学科学院生物医学工程研究所纳米与分子设计实验室合成的新型光敏药物(药物A)的光稳定性；MTT法检测药物A对MGC803细胞的杀伤作用；共聚焦激光显微镜(LSCM)观察药物在MGC803细胞内定位情况；Hoechst染色和流式细胞术(Annexin V/PI双染)检测PDT后细胞死亡方式.结果光照实验表明,药物A对光比较稳定；MTT结果显示,药物A自身对MGC803细胞无暗毒性作用(P＞0.05),单纯光照本身对细胞生长亦无任何影响(P＞0.05),但光照后药物A对MGC803细胞有强烈杀伤作用(P＜0.05),半数致死剂量LD50值为1.74 μmol/L,当药物A浓度为3.12 μmol/L时,细胞存活率为(17.3±1.8)％,浓度继续增加药效不再增加;LSCM观察显示,药物定位于MGC803细胞溶酶体内；Hoechst染色和Annexin V/PI双染表明,细胞死亡方式主要为坏死.结论药物A是一种能够有效杀伤胃癌MGC803细胞的新型光敏药物.
목적 탐토일충신형광민약물대인위암세포MGC803적광동역학치료(PDT)작용급기궤제.방법 채용bleaching광표백법평개중국의학과학원생물의학공정연구소납미여분자설계실험실합성적신형광민약물(약물A)적광은정성；MTT법검측약물A대MGC803세포적살상작용；공취초격광현미경(LSCM)관찰약물재MGC803세포내정위정황；Hoechst염색화류식세포술(Annexin V/PI쌍염)검측PDT후세포사망방식.결과 광조실험표명,약물A대광비교은정；MTT결과현시,약물A자신대MGC803세포무암독성작용(P＞0.05),단순광조본신대세포생장역무임하영향(P＞0.05),단광조후약물A대MGC803세포유강렬살상작용(P＜0.05),반수치사제량LD50치위1.74 μmol/L,당약물A농도위3.12 μmol/L시,세포존활솔위(17.3±1.8)％,농도계속증가약효불재증가;LSCM관찰현시,약물정위우MGC803세포용매체내；Hoechst염색화Annexin V/PI쌍염표명,세포사망방식주요위배사.결론 약물A시일충능구유효살상위암MGC803세포적신형광민약물.
Objective To investigate the treatment efficiency of a new photodynamic therapeutic (PDT) drug synthesized by our laboratory toward MGC803 cells and related mechanisms.Methods Bleaching method was used to evaluate the photostability of drug upon repetitive illumination.MTT assay was used to determine the ability of new drug killing MGC803 cells after PDT.Laser scanning confocal microscopy (LSCM) was applied to investigate the subcellular localization of drug in MGC803 cells (mitochondria and/or lysosomes).Hoechst staining and flow cytometry (Annexin V/PI double-staining)were performed to detect the death mode of MGC803 cells after PDT.Results This new PDT drug had good stability to light irradiation after repetitive illumination.MTT assay showed no cytotoxicity towards MGC803 cells only by drug or only by irradiation(P＞0.05),but intense lethal effect was observed with drug and light combination (P＜0.05).The phototoxicity of medicine increased with the elevation of concentration,the LD50 was 1.74 μmol/L,and reaching plateau at the concentration of 3.12 μmol/L,even incr easing the concentration.LSCM found that drug localized in lysosomes of MGC803 cells.Hoechst staining showed that the death mode of cells was mainly necrosis and Annexin V/PI doublestaining proved this result further.Conclusion This new PDT drug is an effective PDT sensitizer for MGC803 cells and the death mode of cells is mainly necrosis.