靶向沉默ADAM10基因对心肌细胞N-cadherin加工的影响

Silencing of ADAM10 gene by siRNA inhibits the ADAMs cleavage of N-cadherin in cardiac myocytes

目的:研究心肌细胞中解整合素金属蛋白酶家族(ADAMs)成员ADAM10在神经型钙黏蛋白(N-cadherin)底物加工过程中的重要性,为探求该加工途径在维持心肌组织结构形态和完整性中的作用打下基础。方法:将质粒sc-270165 ADAM10 siRNA(r)转染入大鼠心肌细胞(H9c2),建立ADAM10稳定沉默的心肌细胞株。通过Western blotting检测心肌细胞N-cadherin及其C末端片段(CTF)的蛋白表达,流式细胞术检测心肌细胞表面N-cadherin的表达。利用黏附实验检测其对细胞黏附能力的影响。结果:与阴性对照组相比,ADAM10基因沉默组心肌细胞N-cadherin蛋白表达提高,CTF蛋白表达减少;细胞表面N-cadherin的表达增多,心肌细胞黏附能力提高。结论:心肌细胞中ADAM10在N-cadherin的加工水解过程中可能发挥了作用。

AIM：To study the role of a disintegrin and metalloproteinase domain 10 protein（ADAM10） in shedding of neural cadherin（N-cadherin）. METHODS：The siRNA expression vector sc-270165 ADAM10 siRNA（r） targeting the mRNA of ADAM10 was transfected into H9c2 cells. The expression of N-cadherin and its C-terminal fragment（CTF） was detected by Western blotting and flow cytometry. The adhesive ability of the transfected cells was analyzed. RESULTS：The protein level of N-cadherin was increased and the CTF was decreased after ADAM10 siRNA transfection. The accumulation of N-cadherin on the surface of the transfected cells was observed. The adhesive ability of the transfected cells was also significantly increased as compared with control group. CONCLUSION：ADAM10 play a crucial role in shedding of N-cadherin, which may contribute to structural cardiac remodeling.