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目的:探讨光敏剂癌光啉(PsD-007)对肝癌细胞HepG2的体外光动力效应及其机制。方法:将HepG2细胞分为3个光动力处理组,分别以10,20,30μg/mL PsD-007孵育细胞,期间均用630 nm波长的可见光照射(光照能量为6.0 J/cm2,时间2.0 min);同时设3个对照组,分别为空白对照,单纯光照组和暗毒组(30μg/mL PsD-007,未光照)。各组细胞于实验24 h后用MTT法检测存活率,并用凋亡试剂盒结合荧光显微镜观察凋亡情况。应用Real-time PCR和Western blot方法检测20,30μg/mL PsD-007 2个光动力处理组和3个对照组细胞caspase-3,caspase-8和p53的基因、蛋白表达水平。结果:与空白对照组比较,3个光动力处理组HepG2细胞存活率均明显降低,且呈浓度依赖性(P<0.05),而单纯光照组和暗毒组细胞存活率均无明显改变(P>0.05)。荧光显微镜观察结果显示,各光动力处理组坏死或晚期的凋亡细胞较3个对照组显著增多。Real-time PCR和Western blot结果显示,2个光动力处理组细胞caspase-3,caspase-8和p53基因、蛋白表达水平较3个对照组明显上调。结论:PsD-007在体外对HepG2细胞具有光动力杀伤作用,其机制可能与调控caspase蛋白酶和p53的表达从而诱导细胞凋亡和坏死有关。
Objective: To investigate the in vitro photodynamic effects of photosensitizer cancer photoprin (PsD-007) on HepG2 cells and its mechanism. Methods: HepG2 cells were divided into three groups: PDT group (10,20,30μg / mL PsD-007), irradiation with visible light of 630 nm wavelength (light energy 6.0 J / cm2, time 2.0 min ). At the same time, three control groups were set up, which were blank control group, simple light group and dark poison group (30μg / mL PsD-007, no light). The survival rate of each group of cells was measured by MTT method 24 h after the experiment, and the apoptosis was observed by the apoptosis kit combined with fluorescence microscope. Real-time PCR and Western blot were used to detect the expression of caspase-3, caspase-8 and p53 mRNA and protein in 20, 30μg / mL PsD-007 2 photodynamic therapy groups and 3 control groups. Results: Compared with the blank control group, the survival rates of HepG2 cells in the three photodynamic therapy groups were significantly decreased (P <0.05), while the cell survival rates of the two groups were not significantly different (P <0.05) > 0.05). Fluorescence microscopy results showed that each photodynamic therapy group necrosis or late apoptotic cells significantly increased compared with the 3 control group. The results of Real-time PCR and Western blot showed that the expression of caspase-3, caspase-8 and p53 genes and proteins in the two photodynamic therapy groups were significantly up-regulated compared with the three control groups. Conclusion: PsD-007 has a photodynamic killing effect on HepG2 cells in vitro. The mechanism of PsD-007 may be related to the regulation of the expression of caspase and p53 to induce apoptosis and necrosis.