细胞之间介质传导的pCEAcd-tk/前药体系的旁观者效应

The bystander effect of suicide gene pCEAcd-tk/predrug system mediated by the intercellular medium

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 study the mechanism of bystander effect of suicide gene pCEAcd tk/predrug system Methods:plasmid pCEAcd tk was extracted and transfected into the SPCA 1 cells by cationic lipid mediated transfection The transfected cells were mixed with untransfected cells(2∶8) and cultured in 96 well plate (3×10 3cells/well) Predrugs such as 5 FC(5μmol/L) and GCV(10μmol/L) were added into the medium and cultured for 5 days Survival rate was measured with MTT assay,so that to observe bystander effect As to determine the passway of bystander effect ,the SPCA 1 cell line (transfected and untransfected, 2∶8)was cultured in low population(1×10 3cells/well)and high population(5×10 3cells/well), and the cytotoxicity to the wild type cells of the medium and lysate of trans feeted cells was analyzed Results: The bystander effect of pCEA cd tk /predrugs was obvious when the cell population was lower, and there were a toxicity to the wild type cells in the cultured medium and the cell lysate solution Conclusion: The passway of bystander effect in pCEAcd tk/predrugs system may be a intercellular medium mediated process