摘要
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 photodynamic effect of photosensitizer PsD-007 on human hepatocellular HepG2 cells in vitro and its mechanism. Methods： HepG2 ceils were divided into three photodynamic treatment groups, which were incubated with 10, 20 and 30 μg/mL PsD-007 respectively, and were all irradiated with visible light at 630 nm （energy density was 6.0 J/cm^2 and exposure time was 2 rain） during incubation.
Meanwhile： three control groups were used, namely, the blank control group, irradiation alone group and dark cytotoxic group （treated with 30 μg/mL PsD-007 alone without irradiation）. Twenty-four hours after experiment, the ceil survival rates of each group were detected by MTT assa）5 and their apoptosis cells were observed by using apoptosis detection kit and fluorescence microscopy. The gene and protein expression of caspase-3, caspase-8 and P53 in cells of the two photodynarnic treatment groups （20 and 30 μg/mL PsD-007） and three control groups were determined by Real-time PCR and Western blot method, respectively.
Results： Compared with the blank control group, the cell survival rates of the three photodynamic treatment groups significantly decreased in a concentration-dependent manner （all P〈0.05）, while those of the irradiation alone group and dark cytotoxic group showed no obvious change （P〉0.05）. The fluorescence microscopic observation revealed that the late apoptotic and necrotic cells remarkably increased in each photodynamic treatment group compared with the three control groups. The Real-time PCR and Western blot results showed that both gene and protein expression levels of caspase-3, caspase-8 and P53 in the ceils of the two photodynamic treatment groups were significantly higher than those of the three control groups.
Conclusion： PsD-007 has photodynamic cytotoxic effect on HepG2 cells in vitro, and the mechanism is probably related to the regulation ofcaspase and p53 expression that thereby induces the cell apoptosis and necrosis.
出处
《中国普通外科杂志》
CAS
CSCD
北大核心
2012年第8期963-967,共5页
China Journal of General Surgery
关键词
肝肿瘤
光化学疗法
癌光啉
细胞凋亡
Liver Neoplasms
Photochemotherapy
Photocarcinorin
Apoptosis
