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目的 本项目研究了国产新型CaM拮抗剂EBB的抗肝癌作用。方法 体外实验表明EBB对人肝癌细胞系 74 0 2和鼠肝癌细胞系H2 2均有明显抑制作用 ,并有剂量依赖关系 ,其IC5 0分别为 3 312 μg/ml和 1 16 7μg/ml,无毒剂量 (IC10 )的EBB与 5 Fu(IC5 0 )联合应用可明显提高H2 2细胞对5 Fu的敏感性 ,5 Fu的IC5 0由 0 75 μg/ml降至 0 15 μg/ml,敏感性提高 5倍。体内实验证明 ,EBB可使腹水型肝癌小鼠动物长期健康存活 (3个月以上 ) ,长期存活率高达 6 4% ,对照组均在 18天左右死亡。EBB的无毒剂量 (5mg·kg 1·d 1)与 5 Fu (2 5mg·kg 1·d 1)联合应用 ,EBB组的长期存活率 (73% )明显高于 5 Fu组 (2 7% ) ,证明EBB对5 Fu的抗肝癌有增敏作用 ,统计学上有显著性。结果 EBB作用机制的研究显示 ,EBB组P5 3蛋白表达水平明显高于对照组 ,提示EBB对P5 3在翻译水平上有较强的上调作用 ;EBB作为拮抗剂使肝细胞内CaM含量下降 ;使增殖的细胞阻断于G2 M期 ,在G0 /G1期间 ,有二倍体峰以及超微结构可见凋亡细胞。结论 CaM拮抗剂EBB具有强抗肝癌作用 ,对 5 Fu有增敏作用 ,对肝癌细胞凋亡有诱导作用 ,它可上调P5 3蛋白表达及下调CaM含量 ,提示它是一种新型的、有前途的化疗药物 ,有进一步研究和开发价值。
Objective This study investigated the anti-hepatocellular carcinoma effect of a new domestic CaM antagonist EBB. Methods In vitro experiments showed that EBB had a significant inhibitory effect on the human hepatocellular carcinoma cell line 74 0 2 and the murine liver cancer cell line H2 2 in a dose-dependent manner with IC 50 values of 3 312 μg / ml and 1 16 7 μg / ml, respectively The combination of EBB with 5 Fu (IC50) at the IC10 dose significantly increased the sensitivity of H2 2 cells to 5 Fu, and the IC 50 of 5 Fu decreased from 0 75 μg / ml to 0 15 μg / ml. The sensitivity Increase 5 times. In vivo experiments show that EBB can make ascites hepatocellular carcinoma mice long-term healthy survival (more than 3 months), long-term survival rate of up to 64%, the control group died in about 18 days. The long-term survival rate (73%) in EBB group was significantly higher than that in 5 Fu group (27%) when combined with 5 Fu (25 mg · kg · d 1) ), EBB on the 5 Fu anti-liver cancer sensitization, statistically significant. Results The mechanism of EBB showed that the expression of P53 protein in EBB group was significantly higher than that in control group, suggesting that EBB had a strong up-regulation effect on P5 3 at translation level; EBB as antagonist decreased intrahepatic CaM content; Proliferating cells were arrested in G2 M phase. During G0 / G1, there were diploid peaks and apoptotic cells in ultrastructure. Conclusions The CaM antagonist EBB has a strong anti-hepatocarcinoma effect and sensitizes 5 Fu. It induces the apoptosis of hepatoma cells. It can up-regulate the expression of P53 and down-regulate the content of CaM, suggesting that it is a new and promising Of chemotherapy drugs, there is further research and development of value.