光动力治疗促进反义bcr-abl寡核苷酸K562细胞核内转染的实验观察  

Photodynamic gene transfection for bcr-abl antisense oligonucleotides on K562 cells in vitro

作　　者：张林[1] 虞乐华[2] 吴南顺[3] 许川山[2] 

机构地区：[1]解放军第三军医大学新桥医院康复理疗科,重庆市400038 [2]重庆医科大学附属第二医院康复理疗科 [3]重庆第九人民医院妇产科,重庆市400070

出　　处：《中国临床康复》2004年第35期8024-8025,i004,共3页Chinese Journal of Clinical Rehabilitation

基　　金：2001年第三军医大学科研基金;全军青年科研基金(01Q102)~~

摘　　要：目的:体外研究光动力治疗(photodynamictherapy,PDT)对b3a2型反义bcr-abl寡核苷酸(assntisenseoligonucleotides,ASO)慢性髓细胞性白血病(chronicmyelogenousleukemia,CML)细胞株K562转染的影响,为PDT合并反义技术应用于CML患者治疗以及体外骨髓净化提供实验基础。方法:半导体激光,波长650nm,剂量9J/cm2垂直照射。光敏剂为血卟啉单甲醚(hematoperphyrinmonomethylether,HMME),采用体外细胞培养技术,荧光显微镜检测荧光标记的反义bcr-abl寡核苷酸在K562细胞中的分布情况。结果:PDT6h后,细胞内荧光物质主要以明亮的点状聚集在胞核和部分胞浆中,而直接转染时,胞内荧光物质则弥散分布于胞浆中。结论:PDT可以使反义bcr-abl寡核苷酸K562细胞转染主要集中于细胞核内。AIM:To investigate the effects of photodynamic therapy(PDT) in vitro on the transfection of b3a2 type antisense oligonucleotides(ASO) bcr-abl chronic myelogenous leukemia(CML) cell strain K562,and provide experimental supports for the application of the combination of PDT and antisense technology on CML patients and bone marrow purging in vitro. METHODS:Semiconductor laser, with the wave length of 650 nm and dosage of 9 J/cm2 was irradiated vertically.The photosensitizer was hematoprophyrin monomethyl ether(HMME),and cultured in vitro.Distribution of ASO bcr-abl in K562 cells labeled with fluorescence was assayed by fluorescence microscope. RESULTS:After 6-hour PDT,cellular fluorescence substances were mostly located in nucleus and partial cytoplasm,and in the presence of direct transfection,cellular fluorescence substances were diffused in cytoplasm. CONCLUSION:PDT can assemble ASO bcr-abl in K562 cells in nucleus.

关 键 词：反义 PDT 转染 寡核苷酸 K562细胞 光动力治疗 胞浆 细胞核 荧光标记 荧光物质 

分 类 号：R733[医药卫生—肿瘤] R730[医药卫生—临床医学]