LincRNA-p21 promotes mesenchymal stem cell migration capacity and survival through hypoxic preconditioning.
LincRNA-p21 promotes mesenchymal stem cell migration capacity and survival through hypoxic preconditioning.
复制标题
LincRNA-p21通过低氧预处理促进间充质干细胞迁移能力和存活
DOI：
10.1186/s13287-018-1031-x
复制
发表时间：
2018-10-25
影响因子：
7.5
通讯作者：
Guo FM
中科院分区：
文献类型：
--
作者：
Meng SS;Xu XP;Chang W;Lu ZH;Huang LL;Xu JY;Liu L;Qiu HB;Yang Y;Guo FM
背景骨髓间充质干细胞(MSCs)在治疗急性呼吸窘迫综合征(ARDS)中具有强大的稳定作用。然而，骨髓间充质干细胞归巢到受损肺组织的效率和存活率仍然有待解决。因此，本研究旨在探讨大基因间质非编码RNA(LincRNA)-p21能否通过体外低氧预适应促进MSC迁移和存活。方法将MSCs分为常氧培养组(20%O2)和缺氧培养组(1%O2)。为明确其作用和机制，引入慢病毒载体介导的LincRNA-p21基因敲除MSCs和缺氧诱导因子-1α抑制剂KC7F2。此外，还通过划痕实验和井间迁移实验分析了MSC的迁移情况。用细胞计数试剂盒8和台盼蓝染色检测MSC的增殖情况。Annexin V-PE/7-AAD染色流式细胞仪检测细胞凋亡。用逆转录-聚合酶链式反应检测LincRNA-p21和HIF-1α的表达，用免疫印迹和酶联免疫吸附试验检测HIF-1α和CXCR4/7蛋白的表达。WB法检测细胞凋亡蛋白caspase-3和裂解caspase-3的表达。结果低氧预适应α较常氧培养组具有更强的迁移能力和存活能力。低氧预适应诱导的骨髓间充质干细胞表达LincRNA-p21、HIF-1α和CXCR4/7(均为趋化因子基质衍生因子-1受体)。相反，shRNA和低氧诱导因子-1α抑制剂KC7F2阻断LincRNA-p21可抑制低氧预适应诱导的CXCR4/7表达上调、细胞迁移和存活。免疫共沉淀法结果显示，低氧预适应通过增加HIF-1α的表达来分离VHL和HIF-1α蛋白。结论低氧预适应可促进间充质干细胞迁移和存活。在体外低氧预适应下，lincRNA-p21通过HIF-1、α/CXCR4和CXCR7途径促进骨髓间充质干细胞迁移和存活。
BackgroundMesenchymal stem cells (MSCs) derived from bone marrow have potent stabilizing effects for the treatment of acute respiratory distress syndrome (ARDS). However, low efficiency and survival in MSC homing to injured lung tissue remains to be solved. Therefore, the aim of this study was to assess whether large intergenic noncoding RNA (LincRNA)-p21 promote MSC migration and survival capacity through hypoxic preconditioning in vitro.MethodsMSCs were cultured and divided into the normoxia culture group (20% O2) and hypoxia culture group (1% O2). To determine roles and mechanisms, lentivirus vector-mediated LincRNA-p21 knockdown of MSCs and hypoxia-inducible factor (HIF-1α) inhibitor KC7F2 were introduced. Additionally, MSC migration was analyzed by scratch test and transwell migration assays. MSC proliferation was tested by cell counting kit-8 and trypan blue dye. Apoptosis was detected by Annexin V-PE/7-AAD stained flow cytometry. Moreover, LincRNA-p21 and HIF-1α mRNA was measured by reverse transcription-polymerase chain reaction, and HIF-1α and CXCR4/7 protein were assayed by western blot (WB) or enzyme-linked immunosorbent assay (ELISA). Apoptosis protein caspase-3 and cleaved-caspase-3 were investigated by WB analysis. Considering interactions between VHL and HIF-1α under LincRNA-p21 effect, co-immunoprecipitation was detected.ResultsHypoxic preconditioning MSC promoted migration capacity and MSC survival than normoxia culture group. MSCs induced by hypoxic preconditioning evoked an increase in expression of LincRNA-p21, HIF-1α, and CXCR4/7(both were chemokine stromal-derived factor-1(SDF-1) receptors). Contrarily, blockade of LincRNA-p21 by shRNA and HIF-1α inhibitor KC7F2 abrogated upregulation of hypoxic preconditioning induced CXCR4/7 in MSCs, cell migration, and survival. Furthermore, co-immunoprecipitation assay revealed that hypoxic preconditioning isolated VHL and HIF-1α protein by increasing HIF-1α expression.ConclusionsHypoxic preconditioning was identified as a promoting factor of MSC migration and survival capacity. LincRNA-p21 promotes MSC migration and survival capacity through HIF-1α/CXCR4 and CXCR7 pathway under hypoxic preconditioning in vitro.
登录
查看更多内容
影响因子：
56.9
作者：
Jaakkola, P;Mole, DR;Ratcliffe, PJ
通讯作者：
Ratcliffe, PJ
影响因子：
64.5
作者：
Huarte M;Guttman M;Feldser D;Garber M;Koziol MJ;Kenzelmann-Broz D;Khalil AM;Zuk O;Amit I;Rabani M;Attardi LD;Regev A;Lander ES;Jacks T;Rinn JL
通讯作者：
Rinn JL
影响因子：
82.9
作者：
Ceradini, DJ;Kulkarni, AR;Gurtner, GC
通讯作者：
Gurtner, GC
影响因子：
11
作者：
Muscari C;Giordano E;Bonafè F;Govoni M;Pasini A;Guarnieri C
通讯作者：
Guarnieri C
RNA在意外的地方：长的非编码RNA在不同的细胞环境中起作用。
DOI：
10.1038/nrm3679
发表时间：
2013-11
期刊：
Nature reviews. Molecular cell biology
影响因子：
--
作者：
Geisler S;Coller J
通讯作者：
Coller J