广东医学
廣東醫學
엄동의학
GUNAGDONG MEDICAL JOURNAL
2015年
9期
1309-1312,1313
,共5页
孙翔%赵建江%贾搏%韩久松%邱小玲%褚洪星%郑相淮
孫翔%趙建江%賈搏%韓久鬆%邱小玲%褚洪星%鄭相淮
손상%조건강%가박%한구송%구소령%저홍성%정상회
人骨髓间充质干细胞%成骨分化%长链非编码RNA
人骨髓間充質榦細胞%成骨分化%長鏈非編碼RNA
인골수간충질간세포%성골분화%장련비편마RNA
hMSCs%osteogenesis%lncRNA
目的：观察人骨髓间充质干细胞（hMSCs）成骨分化过程中长链非编码 RNA（lncRNA）的表达谱变化。方法以 hMSCs 及成骨诱导分化7、14、21 d 后的细胞为研究对象，利用 Agilent lncRNA 芯片技术筛选出成骨诱导分化前后2倍变化幅度的差异 lncRNA，并通过实时荧光定量 PCR（RT - PCR）对芯片结果进行验证。选取筛选结果中表达差异较大的 lncRNA 利用 UCSC Genome Browser 并结合芯片筛选结果分析 lncRNA 邻近蛋白编码基因。结果 hMSCs 经成骨诱导分化7、14、21 d 后，均具有成骨细胞特性；芯片筛选结果表明，与成骨诱导分化前的hMSCs 相比，成骨诱导分化7、14、21 d 后持续表达上调超过2倍的 lncRNA 有433条，下调超过2倍的有232条；选取其中部分 lncRNA，RT - PCR 验证与芯片结果相符合；对 lncRNA 邻近蛋白编码基因分析提示某些 lncRNA（如lncRNA ENST00000585537.1和 lncRNA eHIT00001595）可能在 hMSCs 成骨分化过程中发挥重要作用。结论 hM-SCs 成骨诱导分化前后，lncRNA 表达谱发生显著变化，提示差异表达的 lncRNA 可能与 hMSCs 成骨分化密切相关，由此可进一步解释 hMSCs 成骨分化机制。
目的：觀察人骨髓間充質榦細胞（hMSCs）成骨分化過程中長鏈非編碼 RNA（lncRNA）的錶達譜變化。方法以 hMSCs 及成骨誘導分化7、14、21 d 後的細胞為研究對象，利用 Agilent lncRNA 芯片技術篩選齣成骨誘導分化前後2倍變化幅度的差異 lncRNA，併通過實時熒光定量 PCR（RT - PCR）對芯片結果進行驗證。選取篩選結果中錶達差異較大的 lncRNA 利用 UCSC Genome Browser 併結閤芯片篩選結果分析 lncRNA 鄰近蛋白編碼基因。結果 hMSCs 經成骨誘導分化7、14、21 d 後，均具有成骨細胞特性；芯片篩選結果錶明，與成骨誘導分化前的hMSCs 相比，成骨誘導分化7、14、21 d 後持續錶達上調超過2倍的 lncRNA 有433條，下調超過2倍的有232條；選取其中部分 lncRNA，RT - PCR 驗證與芯片結果相符閤；對 lncRNA 鄰近蛋白編碼基因分析提示某些 lncRNA（如lncRNA ENST00000585537.1和 lncRNA eHIT00001595）可能在 hMSCs 成骨分化過程中髮揮重要作用。結論 hM-SCs 成骨誘導分化前後，lncRNA 錶達譜髮生顯著變化，提示差異錶達的 lncRNA 可能與 hMSCs 成骨分化密切相關，由此可進一步解釋 hMSCs 成骨分化機製。
목적：관찰인골수간충질간세포（hMSCs）성골분화과정중장련비편마 RNA（lncRNA）적표체보변화。방법이 hMSCs 급성골유도분화7、14、21 d 후적세포위연구대상，이용 Agilent lncRNA 심편기술사선출성골유도분화전후2배변화폭도적차이 lncRNA，병통과실시형광정량 PCR（RT - PCR）대심편결과진행험증。선취사선결과중표체차이교대적 lncRNA 이용 UCSC Genome Browser 병결합심편사선결과분석 lncRNA 린근단백편마기인。결과 hMSCs 경성골유도분화7、14、21 d 후，균구유성골세포특성；심편사선결과표명，여성골유도분화전적hMSCs 상비，성골유도분화7、14、21 d 후지속표체상조초과2배적 lncRNA 유433조，하조초과2배적유232조；선취기중부분 lncRNA，RT - PCR 험증여심편결과상부합；대 lncRNA 린근단백편마기인분석제시모사 lncRNA（여lncRNA ENST00000585537.1화 lncRNA eHIT00001595）가능재 hMSCs 성골분화과정중발휘중요작용。결론 hM-SCs 성골유도분화전후，lncRNA 표체보발생현저변화，제시차이표체적 lncRNA 가능여 hMSCs 성골분화밀절상관，유차가진일보해석 hMSCs 성골분화궤제。
Objective To analyze the expression profile variation of long non - coding RNA(lncRNA)during osteogenesis of human marrow - derived mesenchymal stem cells(hMSCs). Methods Osteoblast - induced differentia-tion was perfomed on hMSCs. Using Agilent lncRNA microarray,samples of hMSCs and differentiated hMSCs on the 7th , 14th and 21st day were assayed to identify the target lncRNA,which was altered more than 2 times during osteogenesis of hMSCs. The expression of lncRNA was verified by real time quantification - polymerase chain reaction(RT - PCR). Spe-cific lncRNAs related protein - coding genes using the UCSC Genome Browser combined with microarray results. Results Osteogenic characteristics were observed in induced hMSCs on the 7th ,14th and 21st day. According to microarray analy-sis,433 lncRNAs were constantly significantly up - regulated more than 2 times and 232 lncRNAs constantly significantly down - regulated more than 2 times during the osteogenesis of hMSCs. The results of RT - PCR were consistent with miRNA microarray. Analysis of nearby protein - coding genes prompted that some lncRNAs,such as lncRNA ENST00000585537. 1 and lncRNA eHIT00001595,may play important roles in osteogenesis of hMSCs. Conclusion During osteogenesis of hMSCs,lncRNA expression profiles have significantly changed,these different lncRNAs maybe closely related to the oste-ogenesis of hMSCs and these findings can also further explain the mechanism of osteogenesis of hMSCs.
