CAJ | 학술논문

背景：人类长链非编码RNA是现今的研究热点，已有研究报道其在肿瘤发生中的作用机制，但其在成骨分化方面的机制并不明确。 　　目的：观察人类长链非编码RNA在经骨形态发生蛋白2诱导小鼠C3H10T1/2细胞成骨分化中的作用，并探讨其分子机制。 　　方法：首先进行碱性磷酸酶染色和成骨指标基因检测。对C3H10T1/2细胞在骨形态发生蛋白2诱导下的成骨分化过程长链非编码RNA表达变化进行芯片分析。采用高通量测序比较骨形态发生蛋白2诱导组和未经骨形态发生蛋白2诱导组的表达变化，筛选出表达下降的基因。过表达相应长链非编码RNA后观察对骨形态发生蛋白2诱导成骨分化的影响。 　　结果与结论：骨形态发生蛋白2诱导C3H10T1/2细胞导致碱性磷酸酶活性增加。骨形态发生蛋白2诱导72 h后，碱性磷酸酶、Id1、骨钙素、Runx2、sp7表达上升(P<0.05)。未诱导C3H10T1/2细胞与骨形态发生蛋白2诱导细胞芯片杂交后结果比较，下降达1.5倍的长链非编码RNAs有24条，其中只有AK035085有内含子。与未过表达AK035085的对照组相比，骨形态发生蛋白2诱导72 h后AK035085过表达的C3H10T1/2细胞碱性磷酸酶、Id1、骨钙素、Runx2、sp7表达均下降(P<0.05)。提示骨形态发生蛋白2可刺激C3H10T1/2细胞发生成骨分化，AK035085可能对C3H10T1/2细胞的成骨分化存在抑制作用。
배경：인류장련비편마RNA시현금적연구열점，이유연구보도기재종류발생중적작용궤제，단기재성골분화방면적궤제병불명학。 　　목적：관찰인류장련비편마RNA재경골형태발생단백2유도소서C3H10T1/2세포성골분화중적작용，병탐토기분자궤제。 　　방법：수선진행감성린산매염색화성골지표기인검측。대C3H10T1/2세포재골형태발생단백2유도하적성골분화과정장련비편마RNA표체변화진행심편분석。채용고통량측서비교골형태발생단백2유도조화미경골형태발생단백2유도조적표체변화，사선출표체하강적기인。과표체상응장련비편마RNA후관찰대골형태발생단백2유도성골분화적영향。 　　결과여결론：골형태발생단백2유도C3H10T1/2세포도치감성린산매활성증가。골형태발생단백2유도72 h후，감성린산매、Id1、골개소、Runx2、sp7표체상승(P<0.05)。미유도C3H10T1/2세포여골형태발생단백2유도세포심편잡교후결과비교，하강체1.5배적장련비편마RNAs유24조，기중지유AK035085유내함자。여미과표체AK035085적대조조상비，골형태발생단백2유도72 h후AK035085과표체적C3H10T1/2세포감성린산매、Id1、골개소、Runx2、sp7표체균하강(P<0.05)。제시골형태발생단백2가자격C3H10T1/2세포발생성골분화，AK035085가능대C3H10T1/2세포적성골분화존재억제작용。
BACKGROUND:Long non-coding RNAs (lncRNAs) have became the hot topic in current studies and play an important role in the tumorigenesis. However, lncRNAs involved in the osteoblast differentiation remain poorly reported. OBJECTIVE:To investigate the role of human LncRNAs in osteogenic differentiation of C3H10T1/2 cells induced by bone morphogenetic protein-2 and explore action mechanism. METHODS:The induction of bone morphogenetic protein-2 was validated by alkaline phosphatase staining and the expression of corresponding genes was detected. The lncRNA expression profile was analyzed using the Arraystar lncRNA array in C3H10T1/2 MSCs undergoing early osteoblast differentiation. The expression with or without bone morphogenetic protein-2 induction was compared with high-flux sequencing, and the down-regulated genes were screened. The effect of lncRNA overexpression on osteogenic differentiation of C3H10T1/2 cells induced by bone morphogenetic protein-2 was observed. RESULTS AND CONCLUSION:The bone morphogenetic protein-2 induced C3H10T1/2 cells led to increased alkaline phosphatase activity. After 72 hours of bone morphogenetic protein-2 induction, alkaline phosphatase, Id1,osteocalcin, Runx2, sp7 expression were increased (P<0.05). There were 24 down-regulated lncRNAs identified between bone morphogenetic protein-2 treated and untreated groups, the decrease of expression was 1.5 folds, and among them, only AK035085 contained intron. Compared with control group with no AK03508 expression, over-expression lncRNA AK035085 decreased the expression of alkaline phosphatase, Id1, osteocalcin, Runx2, sp7 (P<0.05). Experimental findings indicate that bone morphogenetic protein-2 induces osteogenic differentiation of C3H10T1/2 cells and AK035085 inhibits the osteogenic differentiation.