RNA干扰Slug基因对HCT116细胞裸鼠移植瘤生长及转移的影响

Effect of Slug knock-down on tumor formation and metastasis of HCT116 cells in nude mice

目的:探讨通过靶向RNA干扰(RNA interference,RNAi)抑制Slug基因的表达,观测对荷瘤裸鼠结直肠癌生长及转移的影响。方法:利用结肠癌细胞株HCT116对24只5周龄裸鼠皮下种植,建立结肠癌裸鼠皮下移植瘤模型,设立空白对照组、阴性对照组及实验组三组,每组8只。分别注射生理盐水、阴性对照质粒及慢病毒载体,观察肿瘤生长情况,绘制肿瘤生长曲线,观察各组间肿瘤生长及淋巴结转移的变化,应用免疫组化、qRT-PCR和Western blot检测Slug基因和蛋白表达情况。结果:Slug基因shRNA实验组与空白对照组和阴性对照组相比,瘤体生长减缓,移植瘤质量明显减小(3.08±0.31 vs 7.37±1.18,7.46±1.16,P<0.01),Slug蛋白表达明显降低(P<0.05);实验组淋巴结阳性率为36.3%(4/11),与阴性对照组77.8%(14/18)及空白对照组68.4%(13/19)相比较(P<0.01)。结论:靶向Slug的RNA干扰可以显著抑制结肠癌裸鼠模型的生长、淋巴结转移以及癌组织中Slug基因蛋白的表达,可能成为结肠癌基因治疗的潜在分子靶点。

Objective：To explore the influence of RNAi mediated Slug silencing on the growth and metastasis of colon cancer in nude mice. Methods： HCT116 colon cancer cells use for 24 five-week-old nude mice implanted subcutaneously,established colon cancer xenograft model in nude mice,all divided into blank control group,negative control group and the experimental group,each group had eight nude mices. All group were injected with saline,negative plasmid and lentivirus vectors respectively. Tumor growth was observed and draw tumor curved growth,changes in tumor growth and lymph node metastasis between the groups were observed,Slug gene and protein expression were detected by immunohistochemistry,qRT-PCR and Western blot analysis. Results： Slug gene shRNA intervention group compared with the control group and negative control group,tumor grew slower,tumor mass was significantly reduced（3. 08 ±0. 31 vs 7. 37 ± 1. 18,7. 46 ± 1. 16,P〈0. 01）,experimental group of lymph node-positive rate was 36. 3%（ 4 /11）,compared to the negative control group 77. 8%（14 /18） and the control group was 68. 4%（13 /19）（P〈0. 01）. Conclusion： Targeted Slug RNA interference can significantly inhibit the growth of colon cancer in nude mice,lymph node metastasis and the expression of the gene protein in cancer tissue,Slug may be a potential molecular target for colon cancer gene therapy.