siRNA沉默COMMD1后对HEK293细胞衰老相关β-半乳糖苷酶活性的影响  

Effect of silencing COMMD1 by si RNA on the activity of β-Galactosidase related to aging of HEK293 cells

作　　者：蒋智文[1,2,3] 陈维春[1,2,4] 郑慧玲[1,2,3] 袁源[1,2,3] 刘新光[1,2,4] 

机构地区：[1]广东医学院衰老研究所,广东东莞523808 [2]广东省医学分子诊断重点实验室,广东东莞523808 [3]广东医学院东莞科研中心,广东东莞523808 [4]广东医学院生物化学与分子生物学研究所,广东湛江524023

出　　处：《广东医学院学报》2015年第4期367-371,共5页Journal of Guangdong Medical College

基　　金：国家自然科学基金(No.30672205;30900739;81170327);广东省自然科学基金(No.7301506;9252402301000002;9152402301000007);广东省高等学校自然科学研究重点项目(No.06Z015);东莞市科技计划项目(No.200810810 1062;2012108102022);广东医学院建博科技创新团队项目(No.STIF201102);广东医学院青年基金项目(No.2XQ11009;2XK14030)

摘　　要：目的在HEK293细胞中筛选能有效沉默铜离子代谢结构域包含体1(COMMD1)基因表达的si RNA片段,并研究si RNA沉默COMMD1后对HEK293细胞衰老的影响。方法用RT-PCR初步筛选能够沉默COMMD1的si RNA干扰片段,用定量PCR和免疫印迹法进一步确认si RNA的干扰效果,然后用衰老相关β-半乳糖苷酶染色试剂盒检测si RNA沉默COMMD1后对HEK293细胞衰老的影响。结果si COMMD1-1对HEK293细胞中COMMD1的沉默效率最高;但si COMMD1-1沉默COMMD1后,未见HEK293细胞的衰老相关β-半乳糖苷酶活性出现显著性改变。结论 si COMMD1-1沉默HEK293细胞中COMMD1的表达后,未能提高HEK293细胞的衰老相关β-半乳糖苷酶活性。Objective To screen the si RNA fragments that can effectively silence the gene expression of Copper Metabolism（Murr1） Domain-Containing 1（COMMD1）, and to study the effect of silencing COMMD1 by si RNA on the aging of HEK293. Methods si RNA interference fragments that can silence COMMD1 were preliminarily screened by RT-PCR, and interfering effect of si RNA was further determined by q PCR and western blotting. Then, cellular senescence-associated β-Galactosidase Staining Kit was used to determine the effect of silencing COMMD1 by si RNA on the aging of HEK293. Results si COMMD1-1 fragment showed the highest silencing efficiency in HEK293 cells; However, after COMMD1 was silenced by si COMMD1-1, the activity of β-Galactosidase related to aging of HEK293 cells showed no significant change. Conclusion si COMMD1-1 fails to improve the activity of β-Galactosidase related to aging of HEK293 cells after silencing the expression of COMMD1 in HEK293 cells.

关 键 词：COMMD1 SIRNA RNA干扰 细胞衰老 

分 类 号：Q786[生物学—分子生物学]