海鞘素简易结构类似物GJ7-1和GJ7-2的抗肿瘤活性及分子靶向研究

目的:研究依照特有DNA靶向性的抗肿瘤海洋天然产物海鞘素(Ecteinascidins,ETs)合成的简易结构类似物GJ7-1和GJ7-2的抗肿瘤活性及分子靶向。方法:MTT法检测0.012 5¹.6μmol/L的GJ7-1、GJ7-2作用72 h对10种体外培养的肿瘤细胞的增殖抑制作用;荧光结合竞争法测定0.011.6μmol/L的GJ7-1、GJ7-2作用72 h对10种体外培养的肿瘤细胞的增殖抑制作用;荧光结合竞争法测定0.01¹00μmol/L的GJ7-1、GJ7-2与DNA的结合情况;流式细胞仪检测0.01、0.1、1μmol/L的GJ7-1、GJ7-2对A549细胞周期(作用24、48 h)的影响;Hochest33342染色和AnnexinⅤ/PI双染检测0.01、0.1、1μmol/L的GJ7-1、GJ7-2对A549细胞凋亡(作用24、48、72 h)的影响。结果:GJ7-1和GJ7-2对10种肿瘤细胞均有增殖抑制作用,但无明显肿瘤类型选择性;浓度增加到100μmol/L时也与DNA无明显结合;仅1μmol/L的GJ7-1、GJ7-2对A549细胞周期有一定的影响,但均不能诱导其明显凋亡。结论:GJ7-1和GJ7-2虽有一定的抗肿瘤活性,但较ETs大幅降低,部分原因是其完全丧失了与DNA的结合活性。

Anti-tumor Activity and Molecular Targeting of Simplified Ecteinascidins Analogues GJT-1 and GJT-2

OBJECTIVE： To study anti-tumor activity and molecular targeting of simplified analogues GJ7-1 and GJ7-2 synthe-sized with anti-tumor marine products Ecteinascidins （ETs） which target to DNA. METHODS： The inhibitory effects of GJ7-1 or GJ7-2 （0.012 5-1.6 μmol/L, incubating for 72 h） on the proliferation of 10 kinds of tumor cells were tested by MTT assay. GJ7-1 or GJ7-2 （0.01-100 μmol/L） binding to DNA was determined by the fluorescent probe competition assay. The effects of 0.01, 0.1 and 1μmol/L GJ7-1 and GJ7-2 on A549 cell cycle （incubating for 24, 48 h） were observed with flow cytometry. The effects of 0.01, 0.1 and 1 μmol/L GJ7-1 and GJT-2 on A549 cell apoptosis （incubating for 24, 48, 72 h） were observed with Hochest33342 staining and Annexin V/PI staining. RESULTS： Both GJ7-1 and GJ7-2 showed anti-proliferation effect on 10 kinds of tumor cells cultured in vitro without obvious tumor type selectivity. 100 μmol/L GJ7-1 and GJ7-2 did not compete for fluorescent probe binding to DNA. 1 μmol/L GJ7-1 and GJ7-2 influenced cell cycle of A549 cells to some extent and didn＇t induce obvious apoptisis. CONCLUSIONS： GJT-1 and GJ7-2 show anti-tumor activity but ETs has decreased, partly because GJ7-1 and GJ7-2 lose their ability of targeting to DNA completely.