慢病毒介导的RNA干扰下调p16的表达对hUC-MSCs衰老的影响

发布时间：2018-04-22 12:33

本文选题：siRNA + p　；参考：《郑州大学学报(医学版)》2016年04期

【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。
[Abstract]:Aim: to down-regulate the expression of p16 gene in human umbilical cord mesenchymal stem cells (hUC-MSCs) by lentivirus-mediated RNA interference and study the relationship between down-regulation of p16 expression and h UC-MSCs senescence. Methods: the experiment was divided into three groups: the 3rd generation of P3H UC-MSCs group, the P3 group of P15 h UC-MSCs group, the P15 group of P15 UC-MSCs group infected with P15 h, and the LV-NC group of P15 h UC-MSCs group (empty virus group). Cell proliferation was detected by CCK-8 assay, senescence cell rate was calculated by 尾 -galactosidase staining, cell cycle was detected by flow cytometry, and the expression of CDK4 mRNA of senescence related gene p16p21p53pCNAsirt1sirt1siclin D1 + CDK4 was detected by flow cytometry. Results the expression of p16 p21 and p53 mRNA in LV-sip 16 group was lower than that in P15 group, but the expression of pCNAsirt1 sirt2Cyclin D1 + CDK4 mRNA was higher than that of P15 group (P < 0.05). Conclusion: down-regulating the expression of p16 gene in P15 h UC-MSCs can delay and improve the aging of h UC-MSCs.
【作者单位】：郑州大学生命科学学院干细胞研究室;郑州大学第一附属医院干细胞研究室;
【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
【分类号】：R329.2

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