摘要
【目的】研究新型拓扑异构酶Ⅱ抑制剂PPO-1对多种肿瘤细胞株的抑制作用，并初步探讨其作用机制。[方法]MTY法检测PPO-1的体外抗肿瘤活性；光学显微镜观察PPO-1作用后肿瘤细胞形态结构的变化；琼脂糖凝胶电泳分析PPO-1对K562细胞染色体DNA断裂的影响；RT—PCR法检测不同浓度PPO-1（1．0，2．0，4．0μmol／L）对K562细胞caspase-3、topoⅡα／、topoⅡβmRNA表达的影响。【结果】MTT法检测发现PPO-1对多种肿瘤细胞均有较好的活性，IC50为1．41—5．96μmol／L，Giemsa染色发现其出现了典型的凋亡现象，提取总DNA进行琼脂糖凝胶电泳，呈现显著“DNA梯子”，RT—PCR结果显示PPO-1可上调caspase-3及topoⅡdmRNA的表达。【结论】与阳性对照药依托泊苷相比，PPO-1在体外能够抑制多种人源性肿瘤细胞，其抗肿瘤作用机制可能与（1）caspase途径：上调caspase-3基因表达；（2）上调topoⅡα基因表达，抑制topoⅡα催化活性有关。
[Objective]To study the anti-tumor effect in vitro of PPO-1 which is a novel podophyllotoxin derivative, and the mechanism of its anti-tumor effect. [ Methods ] The stronger anti-tumor activeness of PPO- 1 were investigated by MTY assay in vitro. Effect of PPO- 1 on the tumor cell morphology changes in the structure were observed by optical microscope. The mRNA expression of caspase- 3, topo Ⅱ α, topoⅡ β in K562 cells was semi-quantified by reverse transcription PCR （RT-PCR）. [ Results ] The results of MTY assay showed that PPO-1 had favourable activity to many adherent tumor cells and the IC50 values were from 1.41 to 5.96μmol/L. Giemsa staining showed that PPO-1 evoked typical apoptotic features. Agarose gel electrophoresis showed typical DNA fragmentation pattern （DNA ladder） and confirmed the apoptosis induced by different concentrations of PPO-1. The results showed that caspase-3 and topo Ⅱ mRNA levels increased. [ Conclusion ] Compared with the positive control drug, PPO-1 showed stronger anti-tumor activity against various human tumor cell lines. Anti- tumor mechanism of PPO- 1 may be related with the following channels： it could upregulate caspase-3; PPO-1 could inhibite topo Ⅱ α activity by upregulated topo Ⅱ α.
出处
《武警后勤学院学报（医学版）》
CAS
2015年第7期517-521,共5页
Journal of Logistics University of PAP（Medical Sciences）
基金
国家自然科学基金资助项目(30873363)
武警后勤学院博士启动金(WHTD201304-2)
