酪氨酸激酶受体KIT在小鼠结肠黏膜上皮的表达及其功能  

Expression and function of tyrosine kinase receptor KIT in mouse colonic mucosa epithelium

作　　者：康倩[1] 薛红[1] 孙海梅[1] 李小双[1] 张国权[1] 梁元晶[1] 周德山[1] 

机构地区：[1]首都医科大学基础医学院组织学胚胎学教研室,北京100069

出　　处：《解剖学报》2013年第3期393-396,共4页Acta Anatomica Sinica

基　　金：北京市教育委员会科技计划重点资助项目(KZ201311025001)

摘　　要：目的探讨酪氨酸激酶受体KIT在小鼠结肠黏膜上皮的表达及作用。方法应用Western blotting和RT-PCR技术检测c-kit基因及KIT蛋白在野生型C57BL/6小鼠结肠黏膜上皮的表达;用免疫荧光染色显示野生型C57BL/6小鼠以及乙基亚硝酸钠(ENU)诱变的c-kit基因点突变纯合子小鼠(Wadsm/m)结肠黏膜上皮KIT阳性细胞的部位;腹腔注射5-溴脱氧尿嘧啶核苷(BrdU)(30 mg/kg)观察对比野生型以及Wadsm/m小鼠结肠黏膜上皮BrdU掺入情况及动态变化,分析KIT在结肠黏膜上皮更新过程的作用。结果 RT-PCR检测结果表明,在野生型小鼠结肠黏膜组织表达c-kit基因,Western blotting证明在肠黏膜组织存在KIT蛋白;免疫荧光染色显示,KIT阳性细胞位于结肠黏膜上皮,主要位于肠腺隐窝基底部,但是Wadsm/m小鼠KIT阳性细胞数以及KIT蛋白表达量较野生型小鼠明显减少;BrdU掺入实验发现,Wadsm/m小鼠远端结肠黏膜上皮BrdU摄入和更新速度较野生型小鼠明显减慢。结论结肠黏膜上皮表达KIT蛋白与结肠黏膜上皮的更新密切相关。Objective To investigate the expression and function of KIT in mouse colonic mucosa. Methods Western blotting and RT-PCR techniques were used to detect the expression of c-kit gene and KIT in colonic mucosa in C57BL/6 mice （wild-type）. Immunofluorescence staining was used to examine the distribution of KIT-positive cells in colonic mucosa epithelium of C57BL/6 mice （wild-type） and N-ethyl-N-nitrosourea （ENU）-induced c-kit point mutant homozygous mice （Wads^m/m）. Intraperitoneal injection of 5-bromo-2-deoxyuridin （BrdU, 30 mg/kg） was used to observe and compare the BrdU-positive ceils in distal colonic mucosa epithelium in wild-type and Wads^m/m mice, and to evaluate the effect of KIT on the renewal of colonic mucosa epithelium. Results The result of RT-PCR indicated that c-kit gene was expressed in colonic mucosa. Western blotting showed that KIT was expressed in colonic mucosa. Immunofluorescence staining showed that KIT-positive cells existed in the epithelium of colonic mucosa and were mainly located in the lower part of crypts. BrdU incorporation indicated that the renewal of the epithelium of colonic mucosa in homozygous mice was significantly slower than that in wild-type mice. Conclusion C-kit gene and KIT protein are wildely expressed in the colonic mucosa epithelium and might be involved in the renewal of the colonic epithelium.

关 键 词：KIT 肠上皮更新 结肠黏膜 免疫印迹法 小鼠 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]