1例基因芯片筛选青少年特发性脊柱侧凸骨髓间充质干细胞成脂分化相关差异表达基因

Screening of differentially expressed genes associated with adipogenic differentiation of human bone mesenchymal stem cells originated from adolescent idiopathic scoliosis by genome-wide microarray

[目的]应用基因芯片技术筛选青少年特发性脊柱侧凸(AIS)骨髓间充质干细胞(hBMSCs)成脂分化相关的差异表达基因。[方法]收集AIS及同性别年龄正常对照骨髓标本,密度梯度离心法分离MSCs并培养传代,取第3代MSCs进行成脂诱导2周后,提取各标本的总RNA,反转录成cDNA,并以两种荧光Cy5和Cy3标记后作为探针与人类全基因组表达谱基因芯片杂交,扫描芯片荧光信号图像后用基因图像分析软件进行数字化处理和分析。[结果]筛选出显著表达差异基因1 380条,其中表达上调基因874条,表达下调基因506条。[结论]AIS来源MSCs成脂分化相关差异表达的1 380条基因可能参与了AIS的发生和发展。

[ Objective ] To screen the differentially expressed genes associated with adipogenic differentiation of human bone mesenchymal stem cells（MSCs） originated from adolescent idiopathic scoliosis（AIS） by genome-wide microarray. [ Method ] Bone marrow aspirates were obtained from AIS and normal control, then MSCs were separated from bone marrow by the den- sity gradient centrifugution method and cultured primarily to third-passage cells. The P3 cells were adipogenic differentially in- duced for 2 weeks,which were used for this experiment. The total RNA was extracted from AIS and normal control adipogenic differentially induced cells and purified. The eDNA was obtained by reverse transcription polymerase chain reaction （RT-PCR） and then labeled with Cy5 and Cy3 fluorescence probes, which were hybridized with genome-wide microarray. The signal images were scanned and analyzed by image analysis software. [ Result] A total of 1 380 differentially expressed genes were identified by genome-wide microarray, in which 874 genes were up-regulated ,and 506 were down-regulated. [ Conclusion] The 1380 differentially expressed genes associated with adipogenic differentiation of MSCs originated from AIS may be involved in the occurrence and development of AIS.