低氧预处理人胎盘绒毛膜间充质干细胞环状RNA筛选(1)
[摘要] 目的 探讨环状RNA在人胎盘绒毛膜间充质干细胞低氧预处理和常氧培养中表达谱的差异。 方法 利用芯片技术检测3例低氧预处理人胎盘绒毛膜间充质干细胞及其相应对照的常氧培养细胞的环状RNA表达，分析两者差异表达的环状RNA及其结合miRNA。 结果 在检测的13 617条环状RNA中，有分析结果的环状RNA 12 114条，低氧和常氧培养人胎盘间充质干细胞差异表达环状RNA共102条，其中低氧中表达上调的85条，下调的17条(倍数变化>1.5倍且P < 0.05)，而表达差异倍数变化大于2倍以上的环状RNA有27条，且均为上调表达。每个环状RNA预测到5个结合miRNA。 结论 低氧预处理使得人胎盘绒毛膜间充质干细胞的环状RNA表达谱发生了显著变化，这些环状RNA可能和低氧预处理有关。
[关键词] 低氧；人胎盘绒毛膜间充质干细胞；环状RNA；表达谱
[中图分类号] R394.2 [文献标识码] A [文章编号] 1673-7210(2018)06(c)-0009-03
[Abstract] Objective To analyze the expression profile variation of circular RNA (circRNA) between hypoxia preconditioned human placenta chorionic mesenchymal stem cells (H-hpcMSCs) and normoxic preconditioned hpcMSCs (N-hpcMSCs). Methods Human circRNA microarray was performed to detect the difference of circRNA expression between 3 paired specimens of H-hpcMSCs and its N-hpcMSCs. The circRNA differental expression and the predicted miRNA of differentially expressed circRNA were analyzed. Results Of the 13 617 circRNAs， 12 114 circRNAs with analytical results. In H-hpcMSCs， a total of 102 circRNAs showed differentially expressed which have more than 1.5 times variation and significant difference (P < 0.05). Among them， 85 increased and change′s fold more than 1.5 times， while 17 reduced and change′s fold more than 1.5 times， a total of 27 circRNAs showed a greater than 2-fold change and all were up-regulated. Five miRNAs of each with differentially expressed circRNA were predicted according to specific base pairing. Conclusion The circRNA expression profile of H-hpcMSCs changed significantly in comparison with N-hpcMSCs， the circRNA may be associated with the hypoxia precondition.
[Key words] Hypoxia precondition； Human placenta chorionic mesenchymal stem cells； Circular RNA； Expression profile
环状RNA是一类特殊的内源性RNA分子，大量存在于真核生物细胞中，是RNA领域最新的研究热点之一[1-3]。已有研究表明，circRNA参与了多种疾病、衰老等生理病理现象的发生发展过程[4]。目前，circRNA参与调控间充质干细胞生理过程的研究报道较少，而低氧预处理对间充质干细胞中circRNA表达的影响未见报道。因此，本研究预期筛选出低氧预处理人胎盘绒毛膜间充质干细胞差异表达的circRNA，为circRNA参与间充质干细胞的生物学功能提供实验数据。
1 材料与方法
1.1 试剂与仪器
TRIzol试剂(Invitrogen life technologies)，无RNA酶的水(Invitrogen life technologies)，RNase R(Epicentre)，人circRNA芯片v2.0(Arraystar)，Arraystar Supper RNA Labeling试剂盒(Arraystar)，二氧化氮培养箱(Labotect)，基因芯片扫描仪(Agilent)，计算机图像分析系统(Leica)，基因芯片杂交仪(Illumina)
1.2 标本收集和分组
人胎盘绒毛膜间充质干细胞按照已有实验室方法提取并验證为合格的间充质干细胞[5]。本研究中，复温3例不同孕妇胎盘来源的P3代人胎盘绒毛膜间充质干细胞，接种培养至80%融合时消化传代，1∶2分离培养，一瓶置于1%体积分数CO2培养箱中培养，为本次研究的实验组；一瓶置于21%体积分数CO2培养箱中培养，为本次研究的对照组。, 百拇医药(孙逊沙 吴洁莹 陈劲松 陆琰 喻秋霞 李)
[关键词] 低氧；人胎盘绒毛膜间充质干细胞；环状RNA；表达谱
[中图分类号] R394.2 [文献标识码] A [文章编号] 1673-7210(2018)06(c)-0009-03
[Abstract] Objective To analyze the expression profile variation of circular RNA (circRNA) between hypoxia preconditioned human placenta chorionic mesenchymal stem cells (H-hpcMSCs) and normoxic preconditioned hpcMSCs (N-hpcMSCs). Methods Human circRNA microarray was performed to detect the difference of circRNA expression between 3 paired specimens of H-hpcMSCs and its N-hpcMSCs. The circRNA differental expression and the predicted miRNA of differentially expressed circRNA were analyzed. Results Of the 13 617 circRNAs， 12 114 circRNAs with analytical results. In H-hpcMSCs， a total of 102 circRNAs showed differentially expressed which have more than 1.5 times variation and significant difference (P < 0.05). Among them， 85 increased and change′s fold more than 1.5 times， while 17 reduced and change′s fold more than 1.5 times， a total of 27 circRNAs showed a greater than 2-fold change and all were up-regulated. Five miRNAs of each with differentially expressed circRNA were predicted according to specific base pairing. Conclusion The circRNA expression profile of H-hpcMSCs changed significantly in comparison with N-hpcMSCs， the circRNA may be associated with the hypoxia precondition.
[Key words] Hypoxia precondition； Human placenta chorionic mesenchymal stem cells； Circular RNA； Expression profile
环状RNA是一类特殊的内源性RNA分子，大量存在于真核生物细胞中，是RNA领域最新的研究热点之一[1-3]。已有研究表明，circRNA参与了多种疾病、衰老等生理病理现象的发生发展过程[4]。目前，circRNA参与调控间充质干细胞生理过程的研究报道较少，而低氧预处理对间充质干细胞中circRNA表达的影响未见报道。因此，本研究预期筛选出低氧预处理人胎盘绒毛膜间充质干细胞差异表达的circRNA，为circRNA参与间充质干细胞的生物学功能提供实验数据。
1 材料与方法
1.1 试剂与仪器
TRIzol试剂(Invitrogen life technologies)，无RNA酶的水(Invitrogen life technologies)，RNase R(Epicentre)，人circRNA芯片v2.0(Arraystar)，Arraystar Supper RNA Labeling试剂盒(Arraystar)，二氧化氮培养箱(Labotect)，基因芯片扫描仪(Agilent)，计算机图像分析系统(Leica)，基因芯片杂交仪(Illumina)
1.2 标本收集和分组
人胎盘绒毛膜间充质干细胞按照已有实验室方法提取并验證为合格的间充质干细胞[5]。本研究中，复温3例不同孕妇胎盘来源的P3代人胎盘绒毛膜间充质干细胞，接种培养至80%融合时消化传代，1∶2分离培养，一瓶置于1%体积分数CO2培养箱中培养，为本次研究的实验组；一瓶置于21%体积分数CO2培养箱中培养，为本次研究的对照组。, 百拇医药(孙逊沙 吴洁莹 陈劲松 陆琰 喻秋霞 李)