自杀基因治疗神经母细胞瘤的实验研究  

Experimental study on suicide gene therapy of neuroblastoma

作　　者：毕迅[1] 王新娟[2] 张金哲[3] 叶蓁蓁[1] 马继东[1] 郝春生[1] 李跃荣 叶辉[1] 

机构地区：[1]首都儿科研究所附属医院外科,北京100020 [2]北京医科大学肿瘤实验室 [3]北京儿童医院外科

出　　处：《中华小儿外科杂志》2000年第3期139-142,共4页Chinese Journal of Pediatric Surgery

摘　　要：目的　探讨自杀基因疗法对神经母细胞瘤的作用 ,为临床应用打下基础。方法　利用含胸苷激酶基因 (tk)与潮霉素磷酸转移酶基因 (hy)的融合基因的真核表达载体及仅含hy的空载体 ,以脂质体为介导 ,将这两种质粒分别转染神经母细胞瘤细胞系SH SY5Y。利用潮霉素药物敏感实验筛选SH SY5Y ,用不同浓度的丙氧鸟苷 (GCV)分别作用于SH SY5Y、SH SY5Y hytk及SH SY5Y hy。 结果　利用潮霉素药物敏感实验筛选SH SY5Y ,12d时SH SY5Y全部死亡的最低浓度为6 0 μg/ml,30 μm丙氧鸟苷可最大限度杀死SH SY5Y hytk。杀伤效应为 96h死亡率 >80 %。仅含有18%的SH SY5Y hytk就可使周围的 2 0 %的SH SY5Y细胞死亡 ,总死亡率为 38%。 结论　 6 0 μg/ml潮霉素是筛选SH SY5Y hytk及SH SY5Y hy阳性克隆的最佳剂量。 30 μM丙氧鸟苷是作用于SH SY5Y hytk的最佳剂量。tk/GCV系统大于 18%SH SY5Y hytk具有旁观者效应 ,可极大地提高疗效。Objective In order to understand the effect of tk suicide gene therapy on neuroblas- toma. Methods Hygromycin phosphotransferase tk fusion gene and hygromycin phosphotransferase carrier were transduced separately into two groups of neuroblastoma cell line. Different ganciclovir (GCV) dosage was given to SH-SY5Y, SH-SY5Y-hy and SH-SY5Y-hytk separately. Results The cytotoxic effects of GCV on SH-SY5Y-hy-tk cells was obvious (IC 50 = 0.03 *!μM) and killed over 80% of cells after 96 hours with the dosage of 30*!μM GCV. The lowest concentration of hygromycin was 60μg/ml, There was no difference in the growth curve of SH-SY5Y, SH-SY5Y-hy and SH-SY5Y-hytk. Conclusions The best dosage of hygromycin is 60*!μg/ml for selecting the SH-SY5Y-hytk. The best dosage of GCV to kill SH-SY5Y-hytk is 30*!μM . SH-SY5Y-hytk has the bystander effect, if it is greater than 18% in the mixture cells of SH-SY5Y-hytk in the tk/GCV system.

关 键 词：神经母细胞瘤 胸苷激酶 基因治疗 自杀基因 

分 类 号：R730.5[医药卫生—肿瘤]