5-氨基乙酰丙酸光动力杀伤人鼻咽癌耐药/非耐药细胞株的实验研究  

Effect of 5-aminolevulinic acid mediated photodynamic therapy on human nasopharyngeal carcinoma cell line CNE2 and CNE2/DDP in vitro

作　　者：杨小民[1] 罗荣城[1] 马红京[2] 李黎波[1] 丁雪梅[1] 严晓[1] 吕成伟[1] 

机构地区：[1]南方医科大学南方医院肿瘤治疗中心,广州510515 [2]解放军534医院肿瘤科,洛阳471003

出　　处：《第三军医大学学报》2007年第11期1046-1049,共4页Journal of Third Military Medical University

基　　金：中国临床肿瘤学科学基金(Y-2005-0013)~~

摘　　要：目的观察5-氨基乙酰丙酸(5-aminolevulinic acid,5-ALA)光动力学效应对体外培养人鼻咽癌细胞株CNE2及其顺铂耐药株CNE2/DDP的生物作用,探讨光动力治疗多药耐药鼻咽癌的可行性。方法对CNE2与CNE2/DDP分别加入不同浓度的5-ALA(0.01、0.05、0.10、0.25、0.5、1.0、2.0、4.0mmol/L)孵育4h,予波长为630nm的半导体激光进行照射,能量密度分别为2、5、10、20J/cm2,继续孵育24h,用四唑盐比色法测定细胞存活率;两株细胞与2.0mmol/L浓度的5-ALA溶液共孵育4h后,用流式细胞仪检测细胞内生性光敏剂原卟啉Ⅸ的荧光强度。结果5-ALA光动力对体外培养的CNE2和CNE2/DDP细胞均有杀伤作用,表现为对5-ALA浓度、激光剂量的量-效依赖关系。激光能量为20J/cm2时,5-ALA对CNE2和CNE2/DDP细胞的半数杀伤浓度分别为0.07mmol/L与0.375mmol/L,二者比较差异显著(P<0.01);不同5-ALA浓度、不同激光剂量条件下CNE2细胞存活率显著低于CNE2/DDP(P<0.05);CNE2和CNE2/DDP的细胞内荧光强度分别为(197.33±1.45)、(106.36±2.31),相差非常显著(P<0.01)。结论5-ALA-PDT对CNE2和CNE2/DDP均有杀伤作用,但CNE2/DDP对其敏感性低于CNE2,表明CNE2/DDP对5-ALA-PDT存在一定程度的交叉抗性,增加5-ALA浓度和/或增大激光剂量可以在一定程度上克服这种抵抗。Objective To investigate the potential use of 5-aminolevulinic acid （5-ALA） for photodynamic therapy （PDT） of nasopharyngeal carcinoma （NPC） cell line CNE2 and CNE2/DDP cultured in vitro. Methods CNE2 and CNE2/DDP cells were incubated in a medium containing 5-ALA （0.01, 0.05, 0.10, 0.25, 0.5, 1.0, 2.0, 4.0 mmol/L） for 4 h and illuminated with different light doses （2, 5, 10, 20 J/cm^2） using a semiconductor laser at 630 nm. After 24-hour incubation with 5-ALA-PDT, the survival rate of CNE2 and CNE2/DDP cells were analyzed by MTY assay. The intracellular fluorescence of PpIX generated by 5-ALA of each cell was determined by flow cytometry after incubation with 2.0 mmol/L 5-ALA for 4 h. Results 5-ALA-PDT could effectively kill CNE2 and CNE2/DDP cells cultured in vitro. The killing degree was positively correlated with the concentration of 5-ALA and the dosage of radiation. Exposed to 20 J/cm^2, the IC50 of CNE2 and CNE2/DDP was 0.07 mmol/L and 0. 375 mmol/L respectively （P 〈0.01 ）. The survival rate of CNE2/DDP was higher than that of CNE2 under the same 5-ALA concentration and radiation dosage （P 〈 0.05）. The intracellular fluorescence of PpIX in CNE2 and CNE2/DDP was （ 197.33 ± 1.45 ） and （ 106.36 ± 2.31 ） respectively （ P 〈 0.01 ）. Conclusion 5-ALA-PDT can effectively kill CNE2 and CNE2/DDP cells cultured in vitro, though CNE2/DDP cells are less sensitive to 5-ALA-PDT than CNE2 ceils. The cross-resistance of CNE2/DDP might be overcome through increasing 5-ALA concentration and radiation dosage.

关 键 词：鼻咽癌 5-氨基乙酰丙酸 光动力学疗法 CNE2 CNE2/DDP 多药耐药 

分 类 号：R73-36[医药卫生—肿瘤] R739.6[医药卫生—临床医学]