中国实用口腔科杂志

中国实用口腔科杂志

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光动力疗法对颌下腺鳞癌细胞A253杀伤效果研究

王月萍，庄德舒，刘    旭，毕良佳   

  1. 哈尔滨医科大学附属第四医院，哈尔滨  150001 
  • 出版日期:2015-12-15 发布日期:2016-01-06
  • 基金资助:

    高等学校博士学科点专项科研基金联合资助课题（20132307110018）

  • Online:2015-12-15 Published:2016-01-06

摘要：

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≤10 μg/mL时，细胞毒性较小，存活率在92%以上。应用光密度值为80 mW/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    10 μg/mL的HMME介导的PDT对颌下腺鳞癌细胞A253有显著的杀伤效果。

关键词: 光动力疗法, 血卟啉单甲醚, 颌下腺鳞癌, 活性氧

Abstract:

Objective    To invesigate the cytotoxicity effect of hematoporphyrin monomethyl ether （HMME）on the human submandibular gland squamous cell carcinoma A253 cells， and the killing effect of HMME-mediated photodynamic therapy（PDT）  on A253 cells cultured in vitro. Methods    The intracellular concentration changes of HMME were evaluated by fluorescence spectrometer. With the changing of irradiation time， the toxicity of HMME on A253 cells in different concentrations and the killing effect of PDT on A253 cells were detected by MTT assay.  Fluorescence microscopy was used to observe the change of reactive oxygen species（ROS） content after PDT. Results    The intracellular HMME in different concentrations increased and peaked at 2 hours. When the concentration of HMME was less than 10 μg/mL，cell toxicity was low and the survival rate was over 92%. The 630 nm laser of 80 mW/cm2 optical density value was used to irradiate the A253 cells， and with the increasing of illumination time， the percentage of cell survival rate was decreased. At 30 seconds， the cell survival rate was reduced to 71.2%. There was a significant difference between the 30 s-treated group and the control group. ROS fluorescent staining showed that the content of ROS in the cells increased obviously after PDT. Conclusion    Our study show that 10 μg/mL HMME-mediated PDT can significantly restrain the growth of A253 cells.

Key words: photodynamic therapy；hematoporphyrin monomethyl ether；submandibular gland squamous cell carcinoma；reactive oxygen species, ROS