灭活CVB1在离体骨骼肌细胞中基因转移的研究

Gene Transfer Efficiency Enhancement by Inactivated Coxsakie B1 Virus in Skeletal Muscle Cells in Vitro

【目的】研究灭活后的柯萨奇病毒B1在原代培养骨骼肌细胞中对报道基因 pCDNA3 LacZ转移效率的影响。【方法】①用聚乙二醇沉淀和蔗糖低温超速离心法提纯并用 β 丙内酯灭活病毒 ;②分别用不同浓度的转铁蛋白聚赖氨酸 (TfpL) +LacZ、灭活CVB1+LacZ以及灭活CVB1+TfpL +LacZ在肌肉细胞进行基因转移 ,以报告基因产物 β 半乳糖苷酶染色阳性的细胞数作为基因转移效率指标。【结果】灭活CVB1+TfpL +LacZ在肌肉细胞中使基因转移效率由TfpL +LacZ组的不到1%提高到 15 %～ 2 0 %。【结论】灭活CVB1也能象腺病毒一样促进基因转移 ,CVB1的嗜肌肉特性为在肌肉组织中研究靶向明确的新载体提供了实验依据。

Objective To study the effect of inactivated Coxsackie virus B1, which was considered to have high affinity to skeletal muscle cells, on gene transfer efficiency of reporter gene, pCDNA3-LacZ, in primary muscle cells. Methods ① Virus was purified by polyethylene sedimentation followed by sucrose supercentrifugation, ② Transferrin-polylysine (TfpL)+LacZ, inactivated CVB1+LacZ and inactivated CVB1+TfpL+LacZ were designed in different concentrations for the use of gene transfer, then, positive cells contained β-galactosidase were calculated as gene transfer efficiency. Results Inactivated CVB1 enhanced gene transfer efficiency from less than 1% in TfpL+LacZ group to 15%～20% per cent in CVB1+ TfpL+LacZ group. Conclusions Like adenovirus, inactivated CVB1 can also increase gene transfer efficiency in muscle cells, which provides experimental basis for constructing new vector targeting to muscle cells.