小干扰RNA靶向沉默诱骗受体3基因抑制小鼠结肠癌转移进展的研究

Study on small interfering RNA inhibiting the tumor development of mice colon cancer metastasis by silenting decoy receptor 3 gene

目的探索小干扰RNA(siRNA)靶向沉默诱骗受体3基因对小鼠结肠癌转移进展的抑制作用,验证下调诱骗受体3表达对结肠癌体内转移模型恶性生物学性状的影响。方法选取无特定病原体级BALB/c裸鼠30只,使用人结肠腺癌细胞系SW480建立人结肠腺癌裸鼠原位种植转移模型,采用随机数字表法分为诱骗受体3 siRNA组、脂质体对照组与空白对照组,每组10只。种植成瘤后,空白对照组每2天腹腔注射0.9%氯化钠注射液,脂质体对照组每2天腹腔注射脂质体+0.9%氯化钠注射液,诱骗受体3 siRNA组每2天腹腔注射诱骗受体3 siRNA+脂质体+0.9%氯化钠注射液,3组均连续注射7次。观察肿瘤的生长和转移情况,检测肿瘤组织内血管内皮生长因子(VEGF)、基质金属蛋白酶2(MMP-2)、D2-40抗体的表达情况,观察肿瘤组织内的微血管密度及细胞凋亡情况。结果诱骗受体3 siRNA组、脂质体对照组、空白对照组瘤体体积分别为(0.0020±0.0009)、(2.2296±0.0959)、(0.3262±0.1782)cm^(3)。3组均未见肝脏及其他器官转移。诱骗受体3 siRNA组肿瘤组织中VEGF、MMP-2和D2-40抗体表达均少于脂质体对照组。透射电子显微镜观察结果显示,诱骗受体3 siRNA组肿瘤细胞细胞膜周围吞饮小泡减少,细胞核和细胞器的结构清晰,肿瘤细胞间新生血管、淋巴管及增生内皮细胞团明显减少。结论诱骗受体3 siRNA可在小鼠体内成功抑制结肠癌转移瘤的进展。

Objective To explore small interfering RNA(siRNA)inhibiting the tumor development of mice colon cancer metastasis by silenting decoy receptor 3(DcR3)gene,in order to verify the influence of down-regulating DcR3 expression on malignant biological traits in colon cancer metastasis model.Methods Totally 30 specific pathogen free BALB/c nude mice were enrolled.Human colon cancer cell line SW480 was used to establish in situ implantation metastasis model of human colon cancer in nude mice.The mice were randomly divided into DcR3-siRNA group,liposome control group and blank control group,with 10 mice in each group.After implantation and tumorigenesis,the blank control group was given 0.9%sodium chloride injection for intraperitoneal injection every 2 d a time,the liposome control group was given liposome+0.9%sodium chloride injection for intraperitoneal injection every 2 d a time,and the DcR3-siRNA group was given DcR3-siRNA+liposome+0.9%sodium chloride injection for intraperitoneal injection every 2 d a time.All the three groups were given 7 times.The growth and metastasis of the tumor were observed,expressions of vascular endothelial growth factor(VEGF),matrix metalloproteinase-2(MMP-2)and D2-40 antibody in tumor tissues were detected,and microvessel density and apoptosis in tumor tissues were observed.Results The tumor volume of DcR3-siRNA group,liposome control group and blank control group was(0.0020±0.0009),(2.2296±0.0959)and(0.3262±0.1782)cm^(3),respectively.No liver or other organ metastasis was found in all the three groups.Expressions of VEGF,MMP-2 and D2-40 antibody in tumor tissues in DcR3-siRNA group were lower than those in liposome control group.Transmission electron microscopy showed that,in DcR3-siRNA group,peritrophagocytic vesicles around cell membrane of tumor cells were significantly reduced,the structure of nuclei and organelles was clear,and the intertumor angiogenesis,lymphatic vessels and proliferative endothelial cell clusters were significantly reduced.Conclusion DcR3-siRNA can successfully inhibit the progress of colon cancer metastasis in mice.