应用血啉甲醚对体外培养鼠皮肤黑色素瘤细胞PDT治疗的实验研究  

Experimental study of melanoma cell following photodynamic therapy using hematoporphyrin monomethyl ether

作　　者：洪洁[1] 张风[1] 杨庆松[1] 贾力蕴[1] 

机构地区：[1]首都医科大学附属北京同仁医院眼科中心,100730

出　　处：《眼科》2005年第6期405-408,共4页Ophthalmology in China

摘　　要：目的 探讨应用国产光敏剂血啉甲醚(hematoporphyrin monomethyl ether,HMME)对鼠皮肤黑色素瘤细胞株 (B16F10细胞株)行光动力疗法(photodynamic therapy,PDT)的效应和机制。设计 实验性研究。研究对象 B16F10细胞 株。 方法 采用HMME作光敏剂,对B16F10细胞用630nm波长的氦氖激光机作光源(功率密度5mW／cm2),以2．5、5．0、 10．0、20．0、40．0、80．0μg／ml的HMME经2．4、3．6、4．8、6．0、7．2J／cm2的激光能量照射后,MTT法测定PDT对B16F10的杀伤率,An- nexin-V／PI双染流式细胞仪测定B16F10的凋亡率。主要指标 吸光度(OD值)、凋亡率。 结果 随光敏剂浓度升高和照光剂 量增加,光动力作用对细胞的相对杀伤率逐渐增大,在低光动力剂量下明显上升,随后上升渐趋缓慢,杀伤曲线呈“S”形。在对 细胞杀伤率约70％的PDT剂量时,细胞死亡机制以凋亡为主,在对细胞杀伤率约99％的过量PDT剂量时,细胞可能损伤过重 发生坏死。结论 应用HMME行PDT治疗对体外培养鼠皮肤黑色素瘤细胞具有明确的杀伤效应,其对细胞的杀伤率具有显著 的剂量-效应关系;在一定PDT剂量范围内细胞死亡机制主要以凋亡为主。Objective To investigate the phototoxic effect and cell death modes of melanoma cell （B16FIO） following photodynamic therapy（PDT） using hematoporphyrin monomethyl ether（HMME）. Design Experimental study. Participants B16F10 cell strain. Methods Melanoma cell （B16F10） was studied to investigate the phototoxic efficacy and cell death modes following PDT with HMME as photosensitizer and 630nm wavelength Helium-Neon laser as light source. Cultured B16F10 was incubated in a medium containing serial concentrations （2.5, 5.0, 10.0, 20.0, 40.0 and 80.0 ug/ml） of HMME and irradiated with different light dosages （2.4, 3.6, 4.8, 6.0 and 7.2 J/cm^2）. Phototoxic effect was analyzed by MTF assay and cell death modes were investigated by flow cytometer using Annexin-V/PI doubling-staining method. Main Outcome Measures Optical density and the rate of apoptosis. Result The phototoxic effect elevated with increasing of concentration of HMME and dosage of light. In the lower dosage of PDT, the photo toxicity increased rapidly and then gradually slowed down to reach a plateau. The curve of phototoxic effect looked like the shape of ＂S＂. Mode of most cell death appeared apoptosis with the dosage of PDT that induced about 70% cell death. In contrast most of them appeared necrosis with overdosage that induced about 99% cell death. Conclusion PDT using HMME has a significant phototoxic effect on melanoma cell in vitro and its killing efficacy appears to be correlated to dosage of photosensitizer and light irradiation. The major cell death mode is apoptosis.

关 键 词：黑色素瘤/治疗 血啉甲醚 光动力疗法 

分 类 号：R739.5[医药卫生—肿瘤]