HMME介导的光动力疗法诱导鼠肝癌MM45T-Li细胞凋亡的实验研究

目的 探讨光动力疗法对鼠肝癌细胞MM45T-Li凋亡的影响.方法 以血卟啉单甲醚(HMME)为光敏剂,630 nm激光为激发光源的光动力疗法作用于鼠肝癌MM45T-Li细胞.细胞分别与2.5、5、10、20.μg/ml的光敏剂孵育4h后,用不同能量密度的激光照射；MTr法检测HMME的暗毒性以及光动力疗法作用24h后的细胞活性；Hoechst细胞核荧光染色法观察HMME介导的光动力疗法对细胞凋亡的影响.结果 在不照光的情况下各组浓度的HMME对细胞活性无明显抑制作用.当HMME浓度为10、20 μg/ml时,PDT对细胞活性的抑制率随能量密度的增加而增加.Hoechst染色观察PDT作用12h后部分细胞出现染色质凝集、核固缩、核碎裂等形态学改变,5.4 J/cm2及7.2 J/cm2组的凋亡率高于对照组(P＜0.05).结论 HMME介导的光动力疗法可以有效地诱导鼠肝癌MM45T-Li细胞凋亡.

Apoptosis induced by HMME mediated photodynamic therapy in murine hepatoma cell line MM45T-Li

Objective To investigate the effects of HMME mediated photodynamic therapy （PDT） on apoptosis of murine hepatoma cell line MM45T-Li. Methods The photodynamic treatment on MM45T-Li cells was performed in vitro using HMME as photosensitizer and 630 nm laser as light source. The cells were incubated with HMME of 2.5, 5, 10 and 20 ug/ml for 4 h respectively,and then irradiated with laser at different dosages. The dark toxicity of HMME and the activity of cells after PDT were evaluated by MTT assay.The effect of PDT on apoptosis were observed by Hoechst fluorescent staining analysis. Results MTT assay showed that there were no significant effects on cell growth in each concentration group of HMME when the cells were not exposed to light. The inhibition ratio of cells increased with laser dosage at HMME concentration of 10 and 20 ug/ml. Chromatin condensation, nuclear pyknosis and karyorrhexis were observed after PDT by Hoechst fluorescent staining analysis. The apoptosis rate increased in groups of 5.4 J/cm2 and 7.2 J/cm2 compared with that of control （P〈0.05）. Conclusion HMME mediated PDT can efficiently induce the apoptosis of MM45T-Li ceils.