辐射诱导性自杀基因对胰腺癌体内治疗效果的实验观察  

The therapeutical effect of radio-inducible suicide gene on pancreatic cancer cell in vivo

作　　者：刘金龙[1] 刘训良[1] 钱清[1] 杜青[1] 郭仕英[2] 李朝军[2] 苗毅[1] 

机构地区：[1]南京医科大学第一附属医院普外科,江苏南京210029 [2]南京师范大学生命科学学院分子医学实验室,江苏南京210029

出　　处：《肝胆胰外科杂志》2008年第1期36-38,41,共4页Journal of Hepatopancreatobiliary Surgery

基　　金：江苏省卫生厅135重点学科基金资助(苏卫科教[2003]19号)

摘　　要：目的将PC-3胰腺癌细胞种植于balb/c裸小鼠皮下,建立移植瘤模型,观察pAdEgr-1-TK注入后在射线和前药GCV下对胰腺癌的治疗作用。方法PC-3胰腺癌细胞以1×107混以0.1 ml磷酸盐缓冲液(PBS)皮下接种于balb/c裸鼠皮下,每组6只,共4组24只,待肿瘤长至约0.6 cm时,分为4组,以PC-3组作为对照,PC-3/pAdE组注射同量混以PBS的pAdEgr-1,PC-3/pAdET(不放射)和PC-3/pAdET(放射)组将纯化腺病毒pAdEgr-1-TK以1×109 pfu/ml、0.1 ml/只直接注射到治疗组肿瘤体内,1次/d,连续3 d,同时,腹腔内每日注射GCV(25 mg/kg),连续14 d,即分两次间隔48 h行剂量为10 Gy的60Co-γ射线照射,观察各组动物肿瘤生长情况,每3天测量肿瘤体积[(长径×短径2)/2],治疗结束后,处死裸鼠,取瘤体称重,并在光镜下观察肿瘤的病理结果变化。结果4组细胞接种后,balb/c裸小鼠均成瘤,成瘤潜伏期无差异,成瘤率为100%,治疗前(60Co-γ射线照射+GCV),四组动物瘤体大小无差异(P=0.824),治疗后,观察对照组肿瘤生长迅速,治疗组肿瘤生长明显减慢,体积和质量PC-3/pAdET(放射)组均明显小于其他3组(P=0.000)。结论成功建立胰腺癌裸鼠移植瘤模型,体内实验证实Egr-1基因启动子在射线作用下可驱动下游TK自杀基因表达而对胰腺癌起治疗作用。Objective The nude mouse model of pancreatic cancer was established by implanting PC-3 cell subcutaneously into the animal, to observe the therapeutical effect of pAdEgr-1-TK with irradiation and prodrug GCV addition to the cancer cell. Methods 1×10^7 PC-3 cells together with 0.1 ml phosphate buffer solution （PBS） were implanted subcutaneously into 24 nude mice, which were divided equally into 4 groups. By the time when the tumor grew to 0.6 cm, the group PC-3/pAdE was injected with the mixture of PBS and pAdEgr-1, pAdEgr-1-TK was directly injected into the tumor of group PC-3/pAdET （non-irradiated） and group PC-3/pAdET （irradiated） qd for 3 days in the dosage of 1×10^9 pfu/ml×0.1 ml per mouse, meanwhile, GCV was injected into the abdominal cavity （25 mg/kg/day） for 14 days, when the mice would be irradiated by ^60Co-γ in dosage 10Gy and once more 48 hours later. The volumes of the tumor were measured every 3 days （longer diameter x shorter diameter 2/2）. At last, the tumors were taken out of the killed mice to observe the pathologic changes under light microscope. Results Tumor successfully grew in all of the mice without variance during the latent period. No variance existed （P=0.824） before treatment （irradiation and GCV）. After the treatment, the growth of tumor was very fast in the control group but markedly slowed down in the experimental group, while the volume and weight of tumor of group PC-3/pAdET were apparently less than that of the other three. All above-mentioned indicated that TK gene promoted by Egr-1 gene was activated to transcript by γ-rays, and TK suicide gene was highly expressed to inhibit the tumor cells significantly. Conclusion The nude mouse model of pancreatic cancer is successfully established, and the downstream TK suicide gene can be highly expressed activated by the radio-inducible Egr-1 gene promoter, which provide therapy effect in pancreatic cancer in vivo.

关 键 词：胰腺肿瘤 基因治疗 EGR-1启动子 大鼠 

分 类 号：R735.9[医药卫生—肿瘤]