慢病毒介导的RNA干扰下调p16的表达对hUC-MSCs衰老的影响  

Effect of lentivirus-mediated RNAi targeting p16 on aging of hUC-MSCs

作　　者：韩康[1] 马珊珊[1] 朱相展 孟楠[2] 王欣欣[2] 邢衢 黄团结 韩莉[2] 石振庆 杨波[2] 关方霞[1,2] 

机构地区：[1]郑州大学生命科学学院干细胞研究室,郑州450001 [2]郑州大学第一附属医院干细胞研究室,郑州450052

出　　处：《郑州大学学报（医学版）》2016年第4期446-450,共5页Journal of Zhengzhou University(Medical Sciences)

基　　金：国家自然科学基金资助项目81471306;U1404313;河南省科技创新人才计划154200510008;河南省高校科技创新团队支持计划15IRTSTHN022;河南省产学研合作项目142107000008

摘　　要：目的:利用慢病毒(LV)介导的RNA干扰下调老龄人脐带间充质干细胞(h UC-MSCs)中p16基因的表达,研究p16表达下调与h UC-MSCs衰老的关系。方法:实验分为第3代(P3)h UC-MSCs组(P3组)、P15 h UC-MSCs组(P15组)、LV-sip16感染P15 h UC-MSCs组(LV-sip16组)和LV-NC感染P15 h UC-MSCs组(空病毒组)。采用CCK-8法检测细胞增殖,β-半乳糖苷酶染色计算衰老细胞率,流式细胞术检测细胞周期及凋亡,qRT-PCR检测衰老相关基因p16、p21、p53、p CNA、sirt1、sirt2、Cyclin D1、CDK4 mRNA的表达。结果:LV-sip16感染能促进P15 h UCMSCs增殖,降低衰老细胞率,促使细胞周期进入S期,抑制h UC-MSCs的早期凋亡(P均<0.05);同时,LV-sip16组细胞中p16、p21、p53 mRNA表达量较P15组降低,而p CNA、sirt1、sirt2、Cyclin D1、CDK4 mRNA表达量升高(P均<0.05)。结论:下调P15 h UC-MSCs中p16基因的表达能够延缓和改善h UC-MSCs的衰老。Aim： To study the effect of lentivirus（ LV）-mediated RNAi targeting p16 on the aging of h UC-MSCs.Methods： There were 4 groups,the 3rd generation h UC-MSCs group（ P3 group）,the 15 th generation h UC-MSCs group（ P15 group）,P15 h UC-MSCs infected by LV-sip16 group（ LV-sip16 group） and P15 h UC-MSCs infected by LV-NC group（ negative control group）. CCK-8 assay was performed to detect cell proliferation,β-galactosidase staining was used to detect aging cells,flow cytometry was used to detect cell cycle and apoptosis,and the expressions of p16,p21,p53,p CNA,sirt1,sirt2,Cyclin D1 and CDK4 mRNA were detected by qRT-PCR. Results： LV-sip16 infection could successfully promote the proliferation of P15 h UC-MSCs,reduce aging cell rate,induce S phase cells,and inhibit the early cell apoptosis（ P 0. 05）; meanwhile,the mRNA expressions of p16,p21,p53 were decreased,while the mRNA expressions of p CNA,sirt1,sirt2,Cyclin D1 and CDK4 were increased（ P 0. 05）. Conclusion： Downregulating the expression of p16 in P15 h UC-MSCs could delay and improve the aging of h UC-MSCs.

关 键 词：siRNA P16 慢病毒 人脐带间充质干细胞 衰老 

分 类 号：Q291[生物学—细胞生物学]