CAJ | 학술논문

背景：最近研究发现众多长链非编码RNA调控干细胞多潜能性和分化。长链非编码RNA AK089560在干细胞多向分化中的表达变化及作用，目前尚不清楚。目的：观察间质干细胞C3H10T1/2成骨分化与成脂分化过程中AK089560的表达。方法：重组人骨形态发生蛋白2诱导间质干细胞C3H10T1/2成骨分化，碱性磷酸酶染色鉴定早期成骨分化。地塞米松、吲哚美辛和胰岛素三因子联合诱导C3H10T1/2成脂分化，油红O染色鉴定成脂分化结果。采用qRT-PCR检测诱导前后不同时间点AK089560表达的变化。采用RNAfold软件预测AK089560二级结构， UCSC基因组浏览器分析AK089560邻近编码蛋白基因，fancyGENE在线软件构件长链非编码RNA与编码基因关系图。结果与结论：C3H10T1/2经成骨诱导后，70%以上细胞碱性磷酸酶染色阳性；成脂分化诱导后，80%以上细胞油红O染色阳性。qRT-PCR结果显示，长链非编码RNA AK089560在成骨分化与成脂分化第2，4，6天表达均明显下降，成骨分化与成脂分化第2，4，6天与相应时间未分化组相比均差异有显著性意义(P<0.05)。生物信息学分析表明，LncRNA AK089560具有复杂茎环结构，与编码基因Sema3a形成sense overlap关系。结果表明AK089560在间质干细胞成骨与成脂分化中下调表达，提示其可能参与调控间质干细胞多向分化。
배경：최근연구발현음다장련비편마RNA조공간세포다잠능성화분화。장련비편마RNA AK089560재간세포다향분화중적표체변화급작용，목전상불청초。목적：관찰간질간세포C3H10T1/2성골분화여성지분화과정중AK089560적표체。방법：중조인골형태발생단백2유도간질간세포C3H10T1/2성골분화，감성린산매염색감정조기성골분화。지새미송、신타미신화이도소삼인자연합유도C3H10T1/2성지분화，유홍O염색감정성지분화결과。채용qRT-PCR검측유도전후불동시간점AK089560표체적변화。채용RNAfold연건예측AK089560이급결구， UCSC기인조류람기분석AK089560린근편마단백기인，fancyGENE재선연건구건장련비편마RNA여편마기인관계도。결과여결론：C3H10T1/2경성골유도후，70%이상세포감성린산매염색양성；성지분화유도후，80%이상세포유홍O염색양성。qRT-PCR결과현시，장련비편마RNA AK089560재성골분화여성지분화제2，4，6천표체균명현하강，성골분화여성지분화제2，4，6천여상응시간미분화조상비균차이유현저성의의(P<0.05)。생물신식학분석표명，LncRNA AK089560구유복잡경배결구，여편마기인Sema3a형성sense overlap관계。결과표명AK089560재간질간세포성골여성지분화중하조표체，제시기가능삼여조공간질간세포다향분화。
BACKGROUND:Recent studies have found that stem cellpluripotency and differentiation is regulated by many long non-coding RNAs (LncRNAs). The expression and effect of LncRNA AK089560 during differentiation of stem cells is unclear. OBJECTIVE:To investigate the expression of LncRNA AK089560 in mesenchymal stem cells C3H10T1/2 undergoing osteogenic and adipogenic differentiation. METHODS:Osteogenic differentiation of mesenchymal stem cells C3H10T1/2 was induced by recombinant human bone morphogenetic protein-2 and evaluated using alkaline phosphatase staining. The adipogenic differentiation of mesenchymal stem cells C3H10T1/2 was induced by three factors (dexamethasone, indomethacin and insulin) and evaluated by oil red O staining. The dynamical expression of LncRNA AK089560 was detected by qRT-PCR assay. The AK089560 secondary structure was predicted using RNAfold software. The relationship between AK089560 and neighboring protein-coding genes was analyzed using UCSC genome browser and visualized by fancyGENE online software. RESULTS AND CONCLUSION:Over 70%of C3H10T1/2 cells were positive for alkaline phosphatase after osteogenic induction and more than 80%of the cells positive for oil red O staining after adipogenic induction. qRT-PCR results showed that the expression of LncRNA AK089560 at days 2, 4, 6 of both osteogenic and adipogenic differentiation was significantly decreased compared with the control group (P<0.05). Bioinformatics analysis showed that there was a stem-loop structure for AK089560 and sense overlap relationship between AK089560 and protein-encoding gene Sema3a. These findings indicate that LncRNA AK089560 expression is reduced during osteogenic differentiation and adipogenic differentiation, showing that AK089560 may be involved in regulating the multi-directional differentiation of stem cells.