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目的探讨血卟啉单甲醚(HMME)对颌下腺鳞癌细胞A253的细胞毒性,同时研究HMME介导的光动力疗法(PDT)对颌下腺鳞癌细胞A253的杀伤效果。方法通过荧光光谱检测不同孵育时间下细胞对HMME的摄取量。设定不同的光照时间对A253细胞进行PDT,使用四甲基偶氮唑蓝(MTT)法检测不同质量浓度的HMME对A253的细胞毒性及PDT的作用效果。荧光显微镜定性检测PDT作用后细胞内活性氧(ROS)含量的改变。结果孵育时间为2 h时,不同浓度的HMME在细胞内蓄积量均达到峰值。HMME质量浓度≤10μg/m L时,细胞毒性较小,存活率在92%以上。应用光密度值为80 m W/cm2的630 nm激光对A253细胞进行PDT,随着光照时间的延长,细胞存活率降低;在30 s时,细胞存活率降低到71.2%,与空白对照组、单纯光照组、单纯光敏剂组相比,差异有统计学意义(P<0.05)。荧光染色结果表明,PDT作用后细胞内ROS含量明显增加。结论 10μg/m L的HMME介导的PDT对颌下腺鳞癌细胞A253有显著的杀伤效果。
Objective To investigate the cytotoxicity of hematoporphyrin monomethyl ether (HMME) on submandibular adenosquamous carcinoma cell line A253 and to study the killing effect of HMME mediated photodynamic therapy (PDT) on submandibular adenosquamous carcinoma cell line A253. Methods Fluorescence spectra were used to detect the uptake of HMME by cells under different incubation time. A253 cells were exposed to different doses of PDT. MTT assay was used to determine the cytotoxicity of AML3 and the effect of PDT. Fluorescence microscopy was used to qualitatively detect the change of intracellular reactive oxygen species (ROS) after PDT treatment. Results When the incubation time was 2 h, different concentrations of HMME peaked in the cytoplasm. HMME mass concentration ≤ 10μg / m L, less cytotoxic, the survival rate of 92% or more. PDT was performed on A253 cells using a 630 nm laser with optical density of 80 mW / cm2. The cell survival rate decreased with the increase of illumination time. The cell viability decreased to 71.2% at 30 s. Compared with the blank control group, Compared with pure light group and simple photosensitizer group, the difference was statistically significant (P <0.05). Fluorescence staining results showed that the intracellular ROS content increased significantly after PDT treatment. Conclusion HMME-mediated PDT at 10 μg / mL has a significant killing effect on A253 submandibular adenocarcinoma cells.