光动力疗法对人肝癌细胞SSMC-7721凋亡及suivivin表达的影响

Effect of Photodynamic Therapy with Hematoporphyrin Derivative in the Apoptosis of Human Hepatocellular Carcinoma Cell line SMMC-7721 and the Expression of Survivin

目的:探讨血卟啉衍生物(hematoporphyrin derivative,HPD)光动力作用(photodynamic therapy,PDT)诱导人肝癌细胞SMMC-7721凋亡的作用及其机制。方法:采用MTT法测定PDT对人肝癌细胞的相对抑制率,并筛选最佳实验参数。应用流式细胞术检测PDT后细胞凋亡率。并用细胞免疫组化法检测survivin及caspase-3蛋白表达水平。结果:当HPD浓度为4mg/L,光照剂量为10J/cm2时,实验效果最佳,PDT光动力组人肝癌细胞SMMC-7721相对抑制率为(64.28±2.73)%,凋亡率达(35.28±1.52)%,与对照组相比,差异有显著意义(P<0.01);PDT后survivin蛋白表达水平显著低于对照组(P<0.05),而caspase-3却高于对照组。结论:血卟啉衍生物光动力作用具有诱导人肝癌细胞SMMC-7721凋亡的生物学效应,其作用机制可能与光动力作用抑制凋亡调控蛋白survivin表达,并直接或间接促进caspase-3的表达,从而促进细胞凋亡。

Objective:To evaluate the effect of photodynamic therapy with hematoporphyrin derivative in the apoptosis of human hepatocellular carcinoma cell line SMMC-7721 and the mathion. Methods: the surviving rate of SMMC-7721 was analyzed by MTT assay after photodynamic therapy.The apoptosis was detected by flow cytometry with AnnexinV-FITC and Propidium Iodide staining. The expression of survivin and caspase-3 of SMMC-7721 cell was determined by immunocytochemistry and RT-PCR assay respectively. Results:The apoptotic rate of human hepatocellular carcinoma cell line SMMC-7721 after photodynamic therapy with hematoporphyrin was significantly increased , which was higher than that in the controls (P<0.05).The expression of survivin protein in SMMC-7721 matoporphyrin group was lower than that in the controls ( P<0.05) and no difference between the controls while the expression of caspase-3 SMMC-7721 matoporphyrin group was higher than the controls.Conclusion: Photodynamic therapy with hematoporphyrin might obviously induce apoptosis of SMMC-7721 cells by inhibiting the translational level of survivin gene while promoting caspase-3.