Jin Xin, Chen Zhongmei, Xiang Li, Luo Qing, Guo Zhenghua, Ding Xionghui, Jin Xianqing
Ministry of Education Key Laboratory of Child Development and Disorders, Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.
General Hospital of Chongqing Iron and Steel Group, Chongqing 400081, P.R. China.
Mol Med Rep. 2014 Jul;10(1):33-8. doi: 10.3892/mmr.2014.2216. Epub 2014 May 7.
Previous studies have shown that disruption of the bone morphogenetic protein (BMP) signaling pathway is an important cause of intestinal cancer in human and animal models. Thus, the purpose of this study was to construct a Balb/C model of colorectal polyps. Pregnant mice at 9.5 days gestation were injected via the tail vein with the pSES-Si BMP4 plasmid bearing a fluorochrome (DsRed) reporter, in order to silence the BMP4 gene in the first generation (F1); this group of mice was named the pSES-BMP4 group Intestinal fluorescence was detected at 1-, 4- and 8-week‑old F1 mice, and reverse transcription-polymerase chain reaction (RT-PCR) and western-blotting assays were used to determine changes in the expression of BMP4. A dissecting microscope and hematoxylin and eosin (H&E) staining were used to observe the cell morphology and appearance of the polyps. DsRed fluorescence was observed in the intestines of 1-week-old F1 mice of the pSES-BMP4 group. BMP4 expression at the mRNA and protein level was reduced in 1-, 4- and 8-week-old F1 mice (P<0.05). However, the level of Smad4 mRNA was only reduced in 8-week-old F1 mice (P<0.05). Multiple hyperplasic polyps emerged in the colon and rectum of the intestines of 4-week-old F1 mice in the pSES-BMP4 group. The size of colorectal polyps increased at 8 weeks, when vessels and polyp pedicles became apparent. In conclusion, silencing of the BMP4 gene using transplacental RNAi injection can induce formation of colorectal polyps in mice.
先前的研究表明,骨形态发生蛋白(BMP)信号通路的破坏是人类和动物模型中肠道癌的重要原因。因此,本研究的目的是构建一个Balb/C结直肠息肉模型。在妊娠9.5天的怀孕小鼠尾静脉注射携带荧光染料(DsRed)报告基因的pSES-Si BMP4质粒,以便在第一代(F1)中沉默BMP4基因;这组小鼠被命名为pSES-BMP4组。在1周龄、4周龄和8周龄的F1小鼠中检测肠道荧光,并使用逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹分析来确定BMP4表达的变化。使用解剖显微镜和苏木精-伊红(H&E)染色观察息肉的细胞形态和外观。在pSES-BMP4组1周龄的F1小鼠肠道中观察到DsRed荧光。在1周龄、4周龄和8周龄的F1小鼠中,BMP4在mRNA和蛋白质水平的表达均降低(P<0.05)。然而,Smad4 mRNA水平仅在8周龄的F1小鼠中降低(P<0.05)。在pSES-BMP4组4周龄的F1小鼠肠道的结肠和直肠中出现了多个增生性息肉。在8周时,结直肠息肉的大小增加,此时血管和息肉蒂变得明显。总之,通过经胎盘RNAi注射沉默BMP4基因可诱导小鼠结直肠息肉的形成。