融合自杀基因对鼻咽癌细胞的体内杀伤作用

Killing Effect of Fusion Suicide Genes on Nasopharyngeal Carcinoma Cells in vivo

[目的]研究融合自杀基因对鼻咽癌细胞的体内杀伤作用。[方法]培养鼻咽癌CNE-2细胞,建立裸鼠荷瘤模型。随机分为6组,每组5只,A组:脂质体包裹质粒瘤内注射瘤内组;B组:脂质体包裹质粒瘤内注射+肿瘤局部照射γ射线(总剂量9Gy)组;C组:脂质体包裹质粒瘤内注射+腹腔注射5氟胞嘧啶(5-Fc)组;D组:脂质体包裹质粒瘤内注射+肿瘤局部照射γ射线(总剂量9Gy)+腹腔注射5-Fc组;E组:肿瘤局部照射γ射线(总剂量9Gy)+腹腔注射5-Fc,以及空白对照组。记录肿瘤体积变化;计算各组肿瘤抑制率,绘制肿瘤生长曲线;取肿瘤组织进行病理观察;鉴定肿瘤组织中CD/UPRT.UL49基因。肿瘤体积变化的比较采用单向方差分析,组间多重比较采用LSD法。[结果]成功建立鼻咽癌裸鼠荷瘤模型,生长曲线显示:5组瘤体积差异有统计学意义(F=720.684,P<0.01),测序结果证实,除E组外,其余各处理组均检测到目的基因,且Western blot检测结果示:B和D组肿瘤组织内自杀基因的表达高于其余各组。病理切片证实肿块为低分化鳞癌,其中D组肿瘤组织在干预作用下出现明显坏死。[结论]E6.BARF1p.CD/UPRT.UL49自杀基因载体联合前药5-Fc在放射线的作用下,可明显抑制鼻咽癌移植瘤的生长,转染该融合自杀基因可增强放疗敏感性,为鼻咽癌的基因—放射治疗开辟了新思路。

Purpose To explore the killing effect of fusion suicide genes in human nasopharyngeal carcinoma （NPC） in vivo. Methods NPC CNE-2 cell line was hosted to establish mice model with tumor which were randomly divided into 6 groups,where there were 5 mice with different interventions. The volumes of tumor were measured and compared,the tumor growth inhibition rates were calculated. The tumor tissue was taken out for analysis by Hematoxylin-Eosin staining,and the existence of CD/UPRT.UL49 was detected in tumor tissue. Analysis of variance was used for the comparison of tumor volume,and the LSD method was used for the multiple comparison among groups. Results Tumor bearing mice models with NPC were successfully established. The tumor sizes of five groups showed significantly difference （F=720.684,P0.01）. The sequencing testing proved the existence of suicide gene in all the treatment groups except group E,and Western blot showed the expression of suicide genes in group B and D was higher than that in other groups. The pathologic section showed low differentiated squamous cell carcinoma in group B,and tumor tissues showed obvious necrosis in group D. Conclusion Under the radiation,suicide genes of E6.BARF1p.CD/UPRT.UL49 combined with 5-Fc obviously suppressed the growth of transplant tumor,and it was proved fusion suicide genes play a role in increasing sensitivity of tumor cells to radiation,which set up new ideas for the gene-radiation therapy for NPC.