GSK-3抑制剂增强携带TRAIL的溶瘤腺病毒对肝癌细胞的杀伤作用

Enhanced Killing Effects on Hepatoma Cells by TRAIL Armed Oncolytic Adenovirus and Glycogen Synthase Kinase-3 Inhibitors

研究携带TRAIL基因的溶瘤腺病毒联合化疗药物GSK-3抑制剂(氯化锂和SB-415286)对人肝癌细胞的体外杀伤作用。采用MTT法检测病毒ZD55-TRAIL联合化疗药物氯化锂和SB-415286对肝癌细胞株HepG-2、BEL-7404以及正常细胞株L02、QSG-7701增殖的抑制作用,并通过结晶紫实验检测进一步证明联合用药的杀伤作用和安全性;利用Hoechst33342染色对肝癌细胞株BEL-7404的细胞凋亡进行形态学观察;通过Western blot检测肝癌细胞HepG-2细胞凋亡信号通路的变化。结果表明:低剂量的氯化锂和SB-415286能显著提高ZD55-TRAIL对肝癌细胞株HepG-2、BEL-7404的杀伤作用,对人体肝正常细胞株L02、QSG-7701无明显的抑制作用。说明GSK-3抑制剂能够解除肝癌细胞对TRAIL的抗性,增强携带TRAIL基因的溶瘤腺病毒对肝癌细胞的杀伤。

The study explores the combinational killing effects of oncolytic adenovirus-mediated TRAIL and chemotherapeutic agent GSK-3 inhibitors（LiCl and SB-415286）against human hepatoma cells in vitro.First of all,MTT assay tests the growth inhibition effects of combinational therapy with LiCl/SB-415286 and ZD55-TRAIL on hepatocellular cell lines HepG-2,BEL-7404 and normal cell lines L02,QSG-7701;Further,the cytopathic effect assay demonstrates the efficacy and the safety of the combination therapy and the apoptosis of BEL-7404 induced by the viro-chemotherapy is observed through Hoechst 33342 staining under fluorescent microscope.Finally,western blot analysis detects the changes in apoptosis signaling pathway of hepatocellular cell line HepG-2.The results show that the killing effects of ZD55-TRAIL on hepatocellular cell lines HepG-2,BEL-7404 are significantly improved when in combination with low dose of LiCl and SB-415286,while the toxicity against normal cell lines L02,QSG-7701 is not apparent.In conclusion,the data demonstrated that the resistence to TRAIL on hepatoma cells can be greatly alleviated by GSK-3 inhibitors,and the killing effects of oncolyitc adenovirus-mediated TRAIL are obviously enhanced.