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目的 探讨腺病毒介导的VEGF启动子调控的单纯疱疹病毒胸苷激酶基因 (HSV tk)系统 (AdVEGF tk)对体外培养的骨肉瘤细胞的选择性杀伤作用。方法 采用 2种重组腺病毒载体AdVEGF tk和AdCMV tk(CMV启动子调控tk基因表达 )体外感染低表达VEGF的成骨细胞系MC3T3 E1和高表达VEGF的骨肉瘤细胞系LM 8,并给予不同浓度的丙氧鸟苷 (GCV)处理 ,用MTT法检测感染细胞的增殖情况。结果 未感染病毒或感染空病毒的MC3T3 E1细胞和LM8细胞对GCV不敏感 ,感染AdCMV tk的MC3T3 E1细胞和LM 8细胞对GCV敏感。与感染空病毒的MC3T3 E1细胞相比 ,感染AdVEGF tk的MC3T3 E1细胞对GCV的敏感性轻度增加 ,用 5 0 μg/mlGCV处理时 ,30 %MC3T3 E1细胞被杀死 ;感染AdVEGF tk的LM8细胞对GCV的敏感性明显增加 ,用 5 0 μg/mlGCV处理时 ,95 %LM 8细胞被杀死 ,产生的细胞毒活性表现为剂量依赖性。结论 腺病毒介导的VEGF启动子 tk/GCV系统对高表达VEGF的骨肉瘤细胞具有杀伤作用。
Objective To investigate the selective killing of osteosarcoma cells cultured in vitro by adenovirus mediated HSV tk gene (HSV tk) mediated by VEGF promoter (AdVEGF tk). Methods Two kinds of recombinant adenoviruses AdVEGF tk and AdCMV tk were used to infect osteosarcoma cell line MC3T3 E1 with low expression of VEGF and osteosarcoma cell line LM 8 with high expression of VEGF in vitro. Ganciclovir (GCV) treatment, using the MTT method to detect the proliferation of infected cells. Results MC3T3 E1 cells and LM8 cells that were not infected or infected with empty virus were insensitive to GCV. MC3T3 E1 cells infected with AdCMV tk and LM 8 cells were sensitive to GCV. MC3T3 E1 cells infected with AdVEGF tk had a slightly increased sensitivity to GCV compared to MC3T3 E1 cells infected with empty virus. 30% of MC3T3 E1 cells were killed when treated with 50 μg / ml of GCV; LM8 infected with AdVEGF tk The sensitivity of cells to GCV was significantly increased. When treated with 50 μg / ml of GCV, 95% of LM 8 cells were killed and the resulting cytotoxicity was dose-dependent. Conclusion Adenovirus-mediated VEGF promoter tk / GCV system has a cytotoxic effect on osteosarcoma cells highly expressing VEGF.