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目的 研究痘苗病毒介导白细胞介素 2 (rVV IL 2 )基因转染C6胶质瘤细胞体外生物学特性。方法 扩增痘苗病毒载体 ,按重复感染度 (MOI) =0 .2 5∶1、0 .5∶1、1∶1、5∶1、10∶1、2 0∶1转染C6胶质瘤细胞 ,分别检测转染后 2、4、6、8、12、2 4h白细胞介素 2 (IL 2 )的表达 ;应用四甲基偶氮唑盐 (MTT)方法对照检测转染细胞 (MOI =1∶1、5∶1、10∶1)及未转染细胞增殖能力的变化。结果 扩增后病毒滴度为 1.0× 10 9pfu/ml;rVV IL 2转染细胞后 2h可有IL 2明显表达 (MOI =1∶1) ,8h后可达 2 0 0U /ml ,MOI <1∶1时IL 2表达量随接种病毒量的增加而提高 ,MOI >1∶1时增加病毒细胞比IL 2表达量无明显增加 ;病毒转染对体外C6胶质瘤细胞增殖能力无影响。结论 rVV IL 2可体外转染C6 胶质瘤细胞并高效表达。
Objective To study the in vitro biological characteristics of vaccinia virus-mediated interleukin 2 (rVV IL 2) transfection of C6 glioma cells. METHODS: The vaccinia virus vector was amplified, and C6 gliomas were transfected with MOI=0.25:1, 0.5:1, 1:1, 5:1, 10:1, and 20:1. The cells were assayed for IL-2 expression at 2, 4, 6, 8, 12, and 24 h after transfection; MTT assay was used to detect transfected cells (MOI = 1:1, 5:1, 10:1) and changes in proliferation of untransfected cells. Results The titer of the virus after amplification was 1.0×10 9 pfu/ml. IL 2 was significantly expressed at 2 h after transfection with rVV IL 2 (MOI =1:1) and reached 200 U/ml after 8 h. MOI <1 The expression of IL-2 increased with the increase of virus inoculation at the time of 1:1. MOI>1:1 did not increase the expression of IL-2 in the virus cells. The virus transfection had no effect on the proliferation of C6 glioma cells in vitro. Conclusion rVV IL 2 can be transfected into C6 glioma cells in vitro and expressed efficiently.