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目的 :探讨融合自杀基因 pCEAcd tk/前药体系的旁观者效应的机理。 方法 :质粒 pCEAcd tk提取后 ,脂质体介导的方法转染到SPCA 1细胞中 ,与未转染的SPCA 1细胞混合 (2∶8)后接种到 96孔培养板中 (每孔 3× 10 3 ) ,加前体药物 5 氟胞嘧啶 [5 Fluorocytesine ,5 FC(5 μmol/L) ]和 [丙氧鸟苷 (Ganciclovir ,GCV(10 μmol/L) ],培养 5天 ,MTT法测定细胞存活率而观察旁观者效应。利用细胞种植密度的稀密和取转导了 pCEAcd tk融合基因的SPCA 1细胞 (处理细胞 )加前体药物的培养液和细胞裂解液对未转染细胞SPCA 1的毒性探讨旁观者效应的传递方式。结果 :融合基因pCEAcd tk/前药体系的旁观者效应在细胞密度较低时更明显 ,处理细胞的细胞培养液具有细胞毒性。结论 :融合自杀基因 pCEAcd tk/前药体系的旁观者效应的传递方式之一是通过细胞之间的某些介质传递。
Objective: To investigate the mechanism of bystander effect of suicide gene pCEAcd tk / prodrug system. METHODS: Plasmid pCEAcd tk was extracted and transfected into SPCA 1 cells by liposome-mediated method and mixed with untransfected SPCA 1 cells (2: 8) and seeded into 96-well culture plates (3 × 10 3) was added and the prodrug 5Fluorocytesine (5 μmol / L) and Ganciclovir (GCV (10 μmol / L) Cell viability and observed bystander effect.Using the density of cell density and the SPCA 1 cells (treated cells) plus the prodrug and the cell lysate transduced with pCEAcd tk fusion gene, 1 was used to investigate the bystander effect delivery.Results: The bystander effect of the fusion gene pCEAcd tk / prodrug system was more pronounced at lower cell densities and cytotoxicity was observed in the cell culture medium treated with pCEAcd tk / prodrug.Conclusion: The fusion suicide gene pCEAcd One of the ways in which the bystander effect of the tk / prodrug system is delivered is through some of the media between the cells.