[1]韩 康),马珊珊),朱相展),等.慢病毒介导的RNA干扰下调p16的表达对hUC-MSCs衰老的影响韩 康,马珊珊,朱相展,孟 楠,王欣欣,[J].郑州大学学报(医学版),2016,(04):446-450.
 HAN Kang),MA Shanshan),ZHU Xiangzhan),et al.Effect of lentivirus-mediated RNAi targeting p16 on aging of hUC-MSCs[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2016,(04):446-450.
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慢病毒介导的RNA干扰下调p16的表达对hUC-MSCs衰老的影响韩 康,马珊珊,朱相展,孟 楠,王欣欣,()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2016年04期
页码:
446-450
栏目:
论著
出版日期:
2016-07-20

文章信息/Info

Title:
Effect of lentivirus-mediated RNAi targeting p16 on aging of hUC-MSCs
作者:
韩 康1)马珊珊1)朱相展1)孟 楠2)王欣欣2)邢 衢1)黄团结1)韩 莉2)石振庆1)杨 波2)关方霞12)#
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
Author(s):
HAN Kang1) MA Shanshan1) ZHU Xiangzhan1) MENG Nan2)WANG Xinxin2) XING Qu1) HUANG Tuanjie1)HAN Li2) SHI Zhenqing1) YANG Bo2) GUAN Fangxia12)
1)Stem Cell Laboratory, School of Life Sciences,Zhengzhou University, Zhengzhou 450001;
2)Stem Cell Laboratory, the First Affiliated Hospital, Zhengzhou University, Zhengzhou 450052
关键词:
siRNA p16 慢病毒 人脐带间充质干细胞 衰老
Keywords:
iRNA p16 lentivirus human umbilical cord derived mesenchymal stem cell aging
分类号:
Q291
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(hUC-MSCs)中p16基因的表达,研究p16表达下调与hUC-MSCs衰老的关系。方法:实验分为第3代(P3)hUC-MSCs组(P3组)、P15 hUC-MSCs组(P15组)、LV-sip16感染P15 hUC-MSCs组(LV-sip16组)和LV-NC感染P15 hUC-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、pCNA、sirt1、sirt2、CyclinD1、CDK4 mRNA的表达。结果:LV-sip16感染能促进P15 hUC-MSCs增殖,降低衰老细胞率,促使细胞周期进入S期,抑制hUC-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pCNA、sirt1、sirt2、CyclinD1、CDK4 mRNA表达量升高(P均<0.05)。结论:下调P15 hUC-MSCs中p16基因的表达能够延缓和改善hUC-MSCs的衰老。
Abstract:
Aim: To study the effect of lentivirus(LV)-mediated RNAi targeting p16 on the aging of hUC-MSCs.Methods: There were 4 groups,the 3rd generation hUC-MSCs group(P3 group), the 15th generation hUC-MSCs group(P15 group), P15 hUC-MSCs infected by LV-sip16 group(LV-sip16 group)and P15 hUC-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,pCNA,sirt1,sirt2,CyclinD1 and CDK4 mRNAwere detected by qRT-PCR.Results: LV-sip16 infection could successfully promote the proliferation of P15 hUC-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 pCNA, sirt1, sirt2, CyclinD1 and CDK4 were increased(P<0.05).Conclusion: Downregulating the expression of p16 in P15 hUC-MSCs could delay and improve the aging of hUC-MSCs.

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备注/Memo

备注/Memo:
*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
#通信作者,女,1969年2月生,博士,教授,研究方向:干细胞与再生医学, E-mail:guanfangxia@126.com
更新日期/Last Update: 2016-07-20