组蛋白去乙酰化酶抑制剂SAHA联合ZD55-TRAIL溶瘤腺病毒对肝癌细胞杀伤作用的研究

研究组蛋白去乙酰化抑制剂SAHA(suberoylanilide hydroxamic acid)联合ZD55-TRAIL对人肝癌细胞株的体外杀伤效果,实验采用MTT法检测SAHA、ZD55-TRAIL以及二者联合对肝癌细胞株Huh-7、Bel-7404、Hep3B以及人正常肝细胞株QSG-7701增殖的抑制作用；利用Hoechst33342染色对联合处理的细胞进行凋亡形态学观察；通过结晶紫实验进一步检测联合用药对肿瘤细胞的杀伤情况；通过Western blot实验在蛋白水平上检测一些凋亡相关蛋白表达情况的变化.实验结果表明:SAHA联合ZD55-TRAIL处理3d后对肝癌细胞株Huh-7的增殖抑制率达到50％,对Hep3B、Bel-7404也达到近40％的杀伤效率,并且联合治疗后对人正常细胞株QSG-7701未见明显的毒副作用.同时结晶紫实验也证明联合治疗对肿瘤细胞的杀伤力强于用SAHA或ZD55-TRAIL单独使用.Western blot实验证明了药物和病毒的联合作用使促凋亡蛋白Bcl-2,Bcl-xL的表达量明显减少,caspase-8和caspase-9蛋白也有相应剪切现象的发生.流式细胞仪同样检测出SAHA和ZD55-TRAIL联合作用对肝癌细胞有很好的杀伤作用.

Study on Targeting Oncolytic Adenovirus that ZD55-TRAIL Linked Suberoylanilide Hydroxamic Acid (SAHA) for Liver Tumor Cells Therapy

The experiment is aimed at studying the anti-tumor effect in liver tumor cells by combining ZD55-TRAIL with a small molecule Suberoylanilide hydroxamic acid(SAHA).First,MTT assay is used to test the growth inhibition effects of single or combination therapy on tumor cell lines Huh7,Bel-7404,Hep3B and human normal cell line QSG-7701.Second,Hoechst33342 is added after administration in tumor cells Huh7,Bel-7404,Hep3B,and 48 hours later cells were observed under fluorescent microscope.Cytopathic effect assay then is conducted to further demonstrate the efficacy of the combination therapy.Results show that the anti-tumor efficacy of ZD55-TRAIL is significantly enhanced in combination with Suberoylanilide hydroxamic acid,while shows no overlapping toxicity against normal cell line QSG-7701.Crystal violet staining also demonstrates that the combination therapy shows better efficacy than single therapy with SAHA or ZD55-TRAIL,respectively.Western blot experiment showed that the protein of pro-apoptosis and the cleavage of both Caspase-8 and Caspase-9 increases by combination therapy on tumor cell lines.Flow cytometry also demonstrates that combination therapy shows better effect.