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目的:探讨腺病毒介导KDR启动子驱动的双自杀基因体系对人胃癌细胞SCG7901的靶向杀伤作用.方法:应用重组腺病毒AdEasy-KDR-CDglyTK体外感染实验组SCG7901细胞株和对照组HepG2细胞株,并给予不同浓度的前药GCV(ganciclovir)和/或5-FC(5-fluorocytosine),观察该体系对SCG7901细胞杀伤效应及其旁观者效应.结果:携带双自杀基因和报告基因(GFP)的重组腺病毒载体,感染复数为100时,95%以上的受感染SCG7901和HepG2细胞中有GFP表达.已转染腺病毒的SCG7901和未转染SCG7901的细胞在细胞生长方面无显著性差异(t=0.224,P=0.823).已转染腺病毒的HepG2细胞和未转染HepG2的细胞在细胞生长方面也无显著性差异(t=0.120,P=0.904).在前药应用下,已转染腺病毒的SCG7901和HepG2细胞表现出对前药不同的敏感性,SCG7901细胞对前药具有较高的敏感性(F=109.43,P=0.000).融合基因的疗效优于单一自杀基因(F=162.22,P=0.000).将感染腺病毒细胞与未感染细胞以不同比例混和培养,观察到该体系明显的旁观者效应.结论:KDR基因启动子可以调控融合基因体系选择性地杀伤人胃癌SCG7901细胞,并存在旁观者效应.
Objective: To investigate the targeted killing effect of adenovirus-mediated dual suicide gene driven by KDR promoter on human gastric cancer cell line SCG7901.Methods: In vitro infection of HepG2 cells with SCG7901 cell line and AdEasy-KDR-CDglyTK recombinant adenovirus The cells were infected with ganciclovir and / or 5-FC (5-fluorocytosine) at different concentrations and the cytotoxicity and bystander effect of this system on SCG7901 cells were observed.Results: The double suicide gene and reporter gene (GFP ) Of recombinant adenovirus vector, GFP expression in more than 95% of infected SCG7901 and HepG2 cells at a multiplicity of 100. There was no significant difference in cell growth between cells transfected with adenovirus SCG7901 and non-transfected SCG7901 (t = 0.224, P = 0.823) .There was no significant difference in cell growth between HepG2 cells transfected with adenovirus and those without HepG2 (t = 0.120, P = 0.904) The transfected adenovirus SCG7901 and HepG2 cells showed different sensitivity to prodrug, and SCG7901 cells had higher sensitivity to prodrug (F = 109.43, P = 0.000). The efficacy of fusion gene was superior to single suicide gene (F = 162.22, P = 0.000). The infected adenovirus was fine The co-culture of cells with uninfected cells in different proportions showed obvious bystander effect.Conclusion: KDR gene promoter can regulate the fusion gene system to selectively kill human gastric cancer SCG7901 cells with bystander effect.