中国组织工程研究2017，Vol.38Issue(5)：766-772,7.DOI:10.3969/j.issn.2095-4344.2017.05.019
长链非编码RNA NR_033474对C3H10T1/2间质干细胞增殖的影响
Effect of long non-coding RNA NR_033474 on proliferation of C3H10T1/2 mesenchymal stem cells
摘要
Abstract
BACKGROUND:Recent studies have found that long non-coding RNAs (lncRNAs) can regulate stem cel proliferation and differentiation. But it is unclear that how lncRNA NR_033474 regulate stem cel proliferation and cel cycle. OBJECTIVE:To investigate the effect of lncRNA NR_033474 on the proliferation and cel cycle regulation in C3H10T1/2 mesenchymal stem cel s after the NR_033474 overexpressed by lentivirus, and to study the possible regulation mechanism of NR_033474 on mesenchymal stem cel s. METHODS:LncRNA NR_033474 was cloned into a lentivirus vector. Lentivirus particles were infected into C3H10T1/2 cel s to upregulate the expression of NR_033474. The NR_033474 expression level was detected by real-time PCR. Compared with the empty lentivirus vector, the proliferation of C3H10T1/2 cel s which overexpressed NR_033474 was detected by cel counting assay and cel cycle was detected using flow cytometry. The expression of cel cycle-associated proteins such as CDK1, Cyclin B1, Cyclin D1 and P53 were detected by western blot assay. RESULTS AND CONCLUSION:Compared with the control group, lncRNA NR_033474 in C3H10T1/2 cel s which overexpressed NR_033474 was increased by about 100 times (P<0.01), and the proliferation of C3H10T1/2 cel s was significantly inhibited after NR_033474 overexpression by lentivirus (P<0.05). In addition, flow cytometry showed that C3H10T1/2 cel s overexpressing NR_033474 were arrested in G2/M phase compared to the control group. Western blot showed that the expression levels of CDK1 and Cyclin B1 were downregulated, while there were no changes in Cyclin D1 and P53 expression. To conclude, these findings suggest that the NR_033474 overexpression significantly inhibits the cel growth of C3H10T1/2 cel s, at least in part, through induction of cel cycle arrest at G2/M phase.关键词
干细胞/分化/间质干细胞/长链非编码RNA/慢病毒感染/细胞增殖/细胞周期/国家自然科学基金分类
医药卫生引用本文复制引用
潘亚琼,戴忠,左长清,汪宗桂..长链非编码RNA NR_033474对C3H10T1/2间质干细胞增殖的影响[J].中国组织工程研究,2017,38(5):766-772,7.基金项目
国家自然科学基金项目(81101357)Funding:the National Natural Science Foundation of China, No.81101357 （81101357）
