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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 effect and mechanism of photodynamic therapy (PDT) on murine skin melanoma cell line (B16F10) by using domestic photosensitizer hematoporphyrin monomethyl ether (HMME). Design experimental research. Research object B16F10 cell line. Methods HMME was used as a photosensitizer, B16F10 cells were irradiated by He-Ne laser with a wavelength of 630 nm (power density 5 mW / cm 2), 2.5, 5.0, 10.0, 20.0, 40.0, The killing rate of PDT to B16F10 was determined by MTT method after exposed to laser energy of 2.4, 3.6, 4.8, 6.0, 7.2 J / cm2 with HMG of 0 μg / ml. The Anxin-V / PI double staining flow cytometry B16F10 apoptosis rate. The main indicators of absorbance (OD value), apoptosis rate. Results With the increase of photosensitizer concentration and illumination dose, the relative killing rate of photodynamic action on cells gradually increased, and then increased at low photodynamic dose, then increased slowly. The killing curve showed “S” shape. The cell death mechanism is predominantly apoptotic at a PDT dose of about 70% cytotoxicity, and the cells may be over-necrotic at overdose doses of about 99% cytotoxicity. Conclusion PDT with HMME has a clear killing effect on melanoma cells cultured in vitro and has a significant dose-effect relationship with cell killing rate. The cell death mechanism is dominated by apoptosis in a certain dose range of PDT.