论文部分内容阅读
目的：应用原代培养的胶质瘤细胞作不研究对象，观察ＡＣＶ对转染了ＨＳＶ┐ｔｋ基因的肿瘤细胞的杀伤作用。方法：表达单纯疱疹病毒胸苷激酶（ＨＳＶ┐ｔｋ）重组逆转录病毒载体ＳＴＫ，经包装细胞ＰＡ３１７包装成重组逆转录病毒，ＮＩＨ３Ｔ３细胞测定病毒滴度，用此病毒感染原代培养的人胶质瘤细胞，以Ｇ４１８选择后获得ＴＫ＋肿瘤细胞。ＭＴＴ法及活细胞动态观察ＡＣＶ对转基因细胞的毒性作用。结果：ＡＣＶ对ｔｋ＋胶质瘤细胞的杀伤作用明显高于ｔｋ－细胞（ｔｋ＋与ｔｋ－细胞存活率差值ｔ检验Ｐ＜０．００２），随着ＡＣＶ浓度的增高，ｔｋ＋胶质瘤细胞的死亡率明显高于ｔｋ－细胞（两直线斜率比较ｔ检验Ｐ＜０．００１）。结论：逆转录病毒载体转染了ＨＳＶ┐ｔｋ基因的原代胶质瘤细胞可选择性的被ＡＣＶ杀死。
OBJECTIVE: To observe the killing effects of ACV on tumor cells transfected with HSV┐tk gene using primary cultured glioma cells as non-research subjects. Methods: Recombinant retroviral vector STK expressing herpes simplex virus thymidine kinase (HSV┐tk) was packaged into recombinant retroviruses by packaging cells PA317, virus titers were measured in NIH3T3 cells, and primary cultured human glia were infected with this virus. The tumor cells were selected with G418 to obtain TK+ tumor cells. MTT assay and live cells were used to observe the toxic effects of ACV on transgenic cells. Results: The killing effect of ACV on tk+ glioma cells was significantly higher than that of tk-cells (tk+ and tk-cell viability difference t test P<0.002). With the increase of ACV concentration, the tk+ glioma cells Mortality was significantly higher than tk-cells (P<0.001 for both straight-line slopes compared to t-test). CONCLUSION: Primary glioma cells transfected with the retroviral vector HSV┐tk gene can be selectively killed by ACV.