摘要
目的：探讨RNAi沉默Rac1基因对结肠癌细胞增殖及裸鼠成瘤的影响。方法①细胞实验：体外培养结肠癌细胞SW480，随机分为空白对照组、阴性对照组、shRNA-Rac1组，阴性对照组、shRNA-Rac1组分别转染空载质粒慢病毒、shRNA-Rac1慢病毒。采用RT-PCR技术、Western blotting法检测各组Rac1 mRNA及其蛋白表达；采用MTT法检测各组转染24、48、72、96 h的细胞增殖情况。②动物实验：取shRNA-Rac1组、阴性对照组转染后细胞分别接种于裸鼠皮下，建立结肠癌裸鼠移植瘤模型（分别记为观察组、对照组），观察裸鼠成瘤及移植瘤生长情况，喂养35天脱臼处死，完整剥离肿瘤组织，称取瘤体质量并测量肿瘤体积。同时检测瘤体 Rac1蛋白表达情况。结果①细胞实验：与阴性对照组比较，shRNA-Rac1组Rac1 mRNA及其蛋白表达明显降低（P均＜0．05）；与阴性对照组比较，shRNA-Rac1组各时间点细胞增殖速度明显降低（P均＜0．05）。②动物实验：对照组癌细胞接种4～8天均可见瘤体形成，观察组接种35天仅2只成瘤，且瘤体形成时间较对照组晚10天。接种35天，观察组、对照组瘤体体积分别为（32．54±43．13）、（948．13±523．50）mm3，瘤体质量分别为（0．023±0．031）、（0．873±0．372）g，两组比较P均＜0．01。与对照组比较，观察组瘤体组织Rac1蛋白阳性表达率明显降低（ P＜0．05）。结论抑制Rac1表达能降低结肠癌细胞增殖速度，抑制裸鼠移植瘤生长。
Objective To investigate the effects of Rac1 gene silenced by RNA interference （RNAi） on cell prolifera-tion and growth of subcutaneous tumor of colon cancer SW480 cells in nude mice.Methods ① Cell experiment： colon cancer SW480 cells cultured in vitro were randomly divided into the blank control group, negative control group, and shRNA-Rac1 group.Recombinant lentivirus virus vector （ NC-shRNA and Rac1-shRNA） were constructed and were respectively transfected into negative control group and shRNA-Rac1 group.The expression levels of Rac1 mRNA and protein were meas-ured by quantitative real-time polymerase chain reaction （ qRT-PCR） , and Western blotting, respectively.Cell proliferation at 24, 48, 72, and 96 h after transfection was determined by MTT.②Animal experiment：cells from shRNA-Rac1 group and negative control group were planted into nude mice to establish mice models of colon cancer （ observation group and control group） .Formulation and growth of the tumor was observed.After the mice were sacrificed by cervical dislocation, tumor tis-sues were removed completely and we measured its weight and size.Meanwhile, Rac1expression was also detected by immu-nohistocytochemistry.Results ①cell experiment：Compared with the blank control group and negative control group, the expression of Rac1 mRNA and protein in the shRNA-Rac1 group decreased significantly （all P〈0.05）.The proliferation a-bility of colon cancer cells was significantly inhibited in the shRNA-Rac1 group at all detecting time points as compared with that of the control group （all P〈0.05）.②Animal experiment：tumor was observed in all cases of the control group 4-8 days＆nbsp;after the transplantation.While in the observation group, tumor was only observed in 2 cases 35 days after the transplanta-tion, and it took 10 days longer to form a tumor than that in the control group.On the 35th day of transplantation, average volume of the tumor in the observation group and control group was （948.13 ±523.50） and （
出处
《山东医药》
CAS
北大核心
2016年第40期1-4,I0001,共5页
Shandong Medical Journal
基金
国家自然科学基金资助项目(81260321)
关键词
结肠癌
RAC1
RNA干扰
裸鼠成瘤
细胞增殖
colon cancer
Rac1
RNA interference
tumor growth in nude mice
cell proliferation
