中文摘要：

为探讨双依他尼酰乙二胺（EDEA）对人耐顺铂卵巢癌细胞（COC1/DDP）的抗肿瘤活性,本实验用CCK-8细胞增殖实验检测顺铂（DDP）及双依他尼酰乙二胺对细胞生长的抑制作用,用流式细胞术检测细胞凋亡率,Western Blot检测细胞内Bcl-2家族凋亡相关蛋白表达,瑞氏染色观察细胞形态。结果显示,EDEA对人正常细胞HEK293、LO2的低毒剂量接近1.2μmol/L,DDP对上述细胞低毒剂量约1.0μmol/L。分别作用COC1/DDP细胞24 h、48 h、72 h后,EDEA的半抑制浓度IC50均为0.67μmol/L,但DDP对应IC50分别为52.9μmol/L、18.1μmol/L和15.2μmol/L。在浓度均为1.0μmol/L时,EDEA作用COC1/DDP细胞48 h后其凋亡率接近44%,而DDP作用此细胞48h后凋亡率仅约4%。与单用1.0μmol/L DDP处理相比,1.0μmol/L EDEA作用48 h后Bcl-2等抗凋亡蛋白下调,Bax等促凋亡蛋白上调,且p-Akt表达被抑制,细胞形态发生凋亡样改变。结论,EDEA对人正常细胞低毒剂量与DDP相当但对COC1/DDP有显著生长抑制作用,且药效远强于相同浓度DDP;EDEA对耐顺铂卵巢癌细胞COC1的生长抑制作用与p-Akt表达抑制和细胞凋亡增强相关。

英文摘要：

The potential inhibition of the growth of cisplatin-resistant ovarian cancer cell line COC1/DDP by ethylenediamine-di-（ethacrynic amide）（EDEA） was investigated.Cell viability was checkedby CCK- 8 assay,apoptosis rate of cells was detected with flow cytometry;expression of Bcl- 2 family proteins was detected with western blotting;morphology of COC1/DDP cells was observed after Wright staining.EDEA showed low toxicity to normal cells with the maximum level of about 1.2 μmol/L on both HEK293 and LO2 cells;DDP showed low toxicity to normal cells with the maximum level of about 1.0 μmol/L.On COC1/DDP,EDEA showed IC50 of 0.67 μmol/L while DDP showed IC50of nearly 52.9 μmol/L,18.1 μmol/L and 15.2 μmol/L after treatment for 24 h,48 h,72 h,respectively.At treatment for48 h,apoptosis rate of COC1/DDP reached 44%when final 1.0 μmol/L EDEA was used,but just about 4%when final1.0 μmol/L DDP was used.After treatment with final 1.0 μmol/L EDEA for 48 h,Bcl- 2- associated anti-apoptotic proteins were down-regulated,while Bax-associated apoptotic proteins were up-regulated;the action of EDEA was associated with the inhibition of p-Akt activation and morphological alterations of cells.Hence,on COC1 /DDP,EDEA has anti-tumor activity much higher than DDP,potentially through induction of apoptosis and the inhibition of p-Akt activation.