靶向诱骗受体3的小干扰RNA抑制人结肠癌裸鼠皮下移植瘤的实验研究  

Inhibitory Effect of DcR3-siRNA Mediated by Liposome on Nude Mice Model of Colorectal Carcinoma

作　　者：张胜行[1] 兰小鹏[1] 易浔飞[1] 

机构地区：[1]南京军区福州总医院检验科,全军检验医学研究所,福建福州350025

出　　处：《生物技术通讯》2009年第3期339-342,共4页Letters in Biotechnology

基　　金：福建省科技厅国际合作重点项目(2004I012)

摘　　要：目的:以脂质体为载体,探讨靶向诱骗受体3(DcR3)的小干扰RNA(siRNA)对人结肠癌SW480细胞裸鼠皮下移植瘤生长的抑制作用。方法:裸鼠背部皮下种植结肠癌SW480细胞,建立结肠癌SW480细胞裸鼠皮下移植瘤模型;针对靶标DcR3基因,利用Dharmacon公司的软件设计合成siRNA序列,通过瘤体局部多点注射脂质体与DcR3siRNA混合物,将DcR3siRNA转染到裸鼠背部皮下移植瘤内,同时设立阴性对照组和空白对照组;观察肿瘤治疗前后体积变化,评价DcR3siRNA对肿瘤的抑制作用;比较肿瘤治疗前后的细胞形态学改变;免疫组织化学及RT-PCR检测DcR3基因表达;原位细胞凋亡检测肿瘤的凋亡。结果:治疗组肿瘤体积明显小于对照组,肿瘤组织内坏死面积增大,细胞凋亡率显著增高,DcR3蛋白表达水平降低;治疗过程中裸鼠生长良好,无明显毒性反应。结论:脂质体介导的DcR3siRNA对裸鼠皮下结肠癌SW480细胞移植瘤有明显的抑制、杀伤作用,细胞凋亡是DcR3siRNA致肿瘤细胞死亡的重要形式。Objective： To investigate the effect of decoy receptor 3（DcR3） small interfering RNA（siRNA） mediated by liposome vector in inhibiting the growth of human cultured cancer cell SW480. Methods： Athymic mice were subcutaneously transplanted with human colon carcinoma SW480 cells to establish transplanted tumor model. To aim directly at target gene DcR3, siRNA sequence was synthesized by Dharmacon company software, then transfected into subcut transplantation tumor in athymic mice, and the negative and blank control group were established. The suppression effect of DeR3 siRNA to tumor was estimated and the change of morphocytology was compared by observing the tumor volume change on pretherapy and posttherapy. The genetic expression of DcR3 was detected by immunohistochemistry and reverse transcriptase polymerase chain reaction, the tumor apoptosis was detected by TUMEL method. Results： DcR3 siRNA can inhibit the growth of human liver cancer cell SW480. The expression of DcR3 mRNA in human cultured cancer cell SW480 was decreased significantly in the gene transfected group as compared to that of control group. Conclusion： DcR3 siRNA mediated by liposome distinctly inhibits and kills subcut transplantation tumor with human colon carcinoma SW480 cells in athymic mice.

关 键 词：结肠癌 基因治疗 SW480细胞 诱骗受体3 小干扰RNA 裸鼠 

分 类 号：R730.2[医药卫生—肿瘤] Q78[医药卫生—临床医学]