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目的 研究逆转录病毒介导的HSV tk/GCV系统对人喉癌细胞的体外杀伤作用。方法 构建携带HSV tk基因的逆转录病毒载体 ,并以该重组逆转录病毒感染人喉癌细胞Hep 2。以G4 18筛选的抗性克隆 (命名为Hep 2 /tk) ,用MTT比色法观察GCV在体外对Hep 2 /tk细胞的杀伤作用。结果 Hep 2 /tk细胞与未转染tk基因的Hep 2 /0、Hep 2细胞相比较 ,三者的生长速度无明显差别。Hep 2 /tk细胞对GCV高度敏感 ,即低浓度GCV(0～ 1mg/L)处理Hep 2 /tk细胞 7d,可将其大部分细胞杀死 ,增加GCV的浓度 (>1mg/L)不能显著增强其对Hep 2 /tk细胞的杀伤作用。 结论 人喉癌细胞Hep 2对HSV tk/GCV系统高度敏感 ,可望为喉癌的治疗提供一种有效的途径
Objective To investigate the in vitro killing effect of retrovirus-mediated HSV tk / GCV system on human laryngeal carcinoma cells. Methods The retroviral vector carrying HSV tk gene was constructed and Hep 2 was infected with the recombinant retrovirus. The resistance of GCV to Hep 2 / tk cells in vitro was observed by MTT colorimetric assay using G4 18 resistant clone (named as Hep 2 / tk). Results There was no significant difference in the growth rate between Hep 2 / tk cells and Hep 2/0 and untransfected tk genes. Hep 2 / tk cells were highly sensitive to GCV, most of them could be killed by Hep 2 / tk cells treated with low concentration of GCV (0 ~ 1 mg / L) for 7 days. The increase of GCV concentration (> 1 mg / L) Enhance its killing effect on Hep 2 / tk cells. Conclusion Human laryngeal carcinoma cell line Hep 2 is highly sensitive to the HSV tk / GCV system and is expected to provide an effective approach for the treatment of laryngeal carcinoma