芯片分析自发恶性转化的大鼠骨髓间充质干细胞的基因表达谱

Gene expression profile of rat mesenchymal stem cell spontaneous transformation determined by cDNA microarray analysis

摘要:

为解析骨髓间充质干细胞(MSCs)自发恶性转化的遗传学基础, 探讨其临床可用性和安全性。采用密度梯度离心法和贴壁筛选法分离大鼠MSCs, 流式细胞仪分析细胞同源性, 体外培养6个月后获得自发恶性转化的MSCs。应用基因芯片分析自发恶性转化的MSCs中差异表达的基因，进一步采用实时定量RT-PCR验证芯片分析结果。MSCs发生自发恶性转化后, 有44条差异表达基因, 其中21条基因表达上调, 23条基因表达下调。经实时定量RT-PCR检测差异表达基因结果与芯片分析结果一致。Wnt、SHH、Notch、TGFb/BMPs等信号转导通路上的若干基因在MSCs自发恶性转化中起到重要作用。

Abstract:

To study the genetic basis of spontaneous transformation of mesenchymal stem cells (MSCs) and clinical application value MSCs were isolated by combining utilization of density gradient centrifugation and adherence screening. After cell homogeneity analysis by flow cytometry, spontaneous transformation MSCs were isolated after six-month in vitro expansion. Then cell total RNA was obtained with Trizol reagent and studied for gene expression profile. Differentially expressed genes between normal MSCs and spontaneous transformation MSCs in cDNA microarray were determined by real-time RT-PCR for validation of the microarray data. Forty-four genes were differentially expressed after spontaneous transformation of MSCs, among which 21 were up-regulated and 23 down-regulated. The result of real-time RT-PCR was in accordance with that of the cDNA microarray. Several genes in SHH, Notch, TGFβ/BMPs signal pathway play an important role in spontaneous transformation of MSCs.