蛋白表达-第三军医大学学报.docVIP

腺病毒介导SCL转基因对ICC缺失模型鼠结肠c-kit蛋白表达的影响 张 林童卫东刘宝华李春穴 (第三军医大学大坪医院野战外科研究所普通外科，重庆400042)摘　要　目的　构建含有SCL 基因的重组腺病毒载体观察其ICC缺失模型鼠结肠c-kit蛋白表达的影响。方法　取ICC缺失模型BALB/c小鼠，经尾静脉注射1.6×109 PFU的重组腺病毒Ad-SCL，Western检测注射后27、15、30d结肠肌条c-kit蛋白的表达结果　Western blot检测显示，在体应用重组腺病毒Ad-SCL后，可以ICC缺失模型鼠结肠肌条的c-kit蛋白的表达7d最明显15d下降、30d恢复正常水平。结论　制备的重组腺病毒Ad-SCL可以上调ICC缺失模型鼠结肠肌条内c-kit蛋白的表达。关键词　腺病毒载体；基因,SCL； The effects of adenovirus-mediated stem cell leukemia gene transfer on the expression of c-kit protein in mice with ICC loss* ZHANG Lin; TONG Wei-dong; LIU Bao-hua** et al. (Department of General Surgery, Institute of Surgery Research, Daping Hospital, Third Military University, Chongqing 400042, China) *Supported by the National Natural Science Foundation of China (NO. **Corresponding author Abstract: Objective To investigate the effects of adenovirus-mediated stem cell leukemia gene transfer on the expression of c-kit protein in mice with ICC loss. Methods 1.6×109 PFU of recombinant adenovirus containing SCL gene was injected into Balb/c mice with ICC loss via the tail vein. Then the protein expression of c-kit were determined using Western blot analysis at 2d、7d、15d and 30d. Results By western blot analysis, expression of c-kit protein in colon of Balb/c mice with ICC loss was detected at 2d after infected with recombinant Ad-SCL, highest at 7d and return to normal levels at 30d.. Conclusion The recombinant adenovirus up-regulated the expression of c-kit protein in colon of Balb/c mice model with ICC loss in vivo. Key words: adenovirus vector; SCL gene; c-kit gene; protein expression 干细胞白血病（stem cell leukemia, SCL）基因最早发现于T淋巴细胞急性白血病细胞内，编码一种螺旋－环－螺旋类转录因子。研究证明SCL是c-kit上游的重要调控基因，具有强烈的促c-kit表达作用。本实验SCL转基因，观察其对c-kit蛋白表达的影响。为转染Cajal间质细胞,观察其对Cajal间质细胞的影响奠定基础。 重组腺病毒Ad-SCL由本课题组。对照阳性腺病毒Ad-Wnt，由重庆医科大学季业伟博士惠赠。B/c小鼠，出生后15d～20d内，雌雄不限，清洁级，由大坪医院野战外科研究所大鼠抗小鼠c-kit单克隆中和抗体ebioscience，HRP大鼠重组腺病毒Ad-SCL介导的SCL转基因ICC缺失模型鼠c-kit蛋白表达 1.2.1 动物模型h内的Balb/c小鼠腹腔内隔天注射c-kit抗体(ACK2)100μg，共5次，于第10d开始试验。采用建立的ICC缺失模型鼠，随机分为3组：组、阳性对