论文部分内容阅读
【目的】检测自杀基因系统单纯疱疹病毒Ⅰ型胸苷激酶/丙氧鸟苷(HSV1-tk/GCV)对小鼠黑色素瘤的杀伤和抑制效应。【方法】将B16/tk(tk+)细胞和未经修饰的B16(tk-)细胞按tk+细胞占总细胞数的0%、10%、20%、40%、80%比例分别进行混合,各组细胞于96孔板中培养,加入丙氧鸟苷(GCV)至15.7μmol/L,采用四甲基偶氮唑盐(MTT)法检测不同tk+细胞比例的HSV1-tk/GCV系统在体外对肿瘤细胞的杀伤率。各组混合细胞分别接种于小鼠腋下,以GCV注射液按100 mg.kg-1.d-1进行治疗,检测瘤体体积、湿质量,测定抑瘤效应。【结果】tk+比例10%以上治疗组,HSV1-tk/GCV系统对黑色素瘤B16细胞均有明显杀伤效应,但旁杀伤作用不明显;从移植瘤体积看,20%以上tk/GCV治疗组肿瘤呈现明显生长抑制状态(P<0.05),以80%tk/GCV治疗组作用最显著(P<0.01);从移植瘤质量看,80%tk/GCV治疗组肿瘤呈现明显生长抑制状态(P<0.01),其他组虽随着tk+细胞比例的增高瘤体平均质量下降,但差异无显著性意义。【结论】HSV1-tk/GCV系统对小鼠黑色素瘤的杀伤和抑制效应均随着B16/tk细胞比例的增高而增强,已建立显示出体外杀伤效应和体内抑瘤活性的小鼠黑色素瘤自杀基因系统。
【Objective】 To detect the killing effect and inhibitory effect of suicide gene HSV1-tk / GCV on mouse melanoma. 【Method】 B16 / tk (tk +) cells and unmodified B16 (tk-) cells were respectively mixed according to the proportion of tk + cells in 0%, 10%, 20%, 40% and 80% Groups of cells were cultured in 96-well plates, GCV was added to 15.7μmol / L, HSV1-tk / GCV system with different tk + cell ratio was detected by MTT assay in vitro Tumor cell killing rate. Each group of mixed cells were inoculated in the armpit of mice respectively, treated with GCV injection 100 mg.kg-1.d-1, volume and wet mass of the tumor were measured, and the anti-tumor effect was determined. 【Results】 The results showed that both HSV1-tk / GCV system and tk + 10% treatment group had obvious killing effect on melanoma B16 cells, but the para-killing effect was not obvious. According to the volume of tumor, more than 20% (P <0.05), and the most significant effect was at 80% tk / GCV treatment group (P <0.01). From the quality of tumor, the tumor of 80% tk / GCV group showed obvious growth inhibition (P < 0.01). Although the average mass of tumor decreased with the increase of tk + cell ratio in other groups, the difference was not significant. 【Conclusion】 Both HSV1-tk / GCV system can kill and suppress melanoma in mice with the increase of the proportion of B16 / tk cells. Murine melanoma mice that have been shown to have anti-tumor activity in vitro and in vivo have been established to kill Gene system.