下调PPP2R5C表达对Jurkat细胞GSK-3β、MDM2、pTEN、TP53通路相关调控基因表达谱的影响

doi:10.3971/j.issn.1000-8578.2015.11.002

下调PPP2R5C表达对Jurkat细胞GSK-3β、MDM2、pTEN、TP53通路相关调控基因表达谱的影响

Effects of PPP2R5C Downregulation on Expression Profile of GSK-3β, MDM2, pTEN and TP53 Signal Pathway Regulating Genes in Jurkat Cells

摘要

摘要: 目的利用基因芯片技术研究RNA干扰下调PPP2R5C对Jurkat细胞中GSK-3β、MDM2、pTEN、TP53通路相关基因表达的影响。方法利用核转染技术将PPP2R5C-siRNA799转导Jurkat细胞，培养48 h后，提取细胞总RNA，纯化mRNA后反转录制备目标序列，与Affymetrix基因表达谱芯片3'IVT杂交，以Affymetrix GeneChip Scanner 3000扫描仪扫描得到原始图像数据，并应用GeneSpringGX 11.0软件进行差异表达谱分析。结果基因芯片分析GSK-3β、MDM2、pTEN、TP53信号通路相关的47个基因，发现全部基因发生了差异表达，其中上调基因有22个，下调基因有25个。明显上调的基因有SNAI1、APC、CDKN1A、ATM、MDM2和pTEN，而明显下调的基因只有GSK-3β。结论下调Jurkat细胞PPP2R5C基因时，在一定程度上选择性调节涉及细胞增殖和凋亡相关的GSK-3β、MDM2、pTEN和TP53信号通路基因的表达，从而抑制Jurkat细胞增殖。

Abstract: Objective To analyze the influence of PPP2R5C downregulation by RNA interference on genes expression related to GSK-3β, MDM2, pTEN and TP53 signaling pathways in Jurkat cells. Methods PPP2R5C-siRNA799 was delivered into Jurkat cells by nucleofection. Total RNA was isolated from treated Jurkat cells at 48h after siRNA delivery. The target sequences were prepared after mRNA purification, and were hybridized with Affymetrix gene expression profile chip 3'IVT. The original image data were collected by Affymetrix GeneChip Scanner 3000 and analyzed the gene expression profile using Gene Spring GX 11.0 software. Results The expression of 47 genes related to GSK-3β, MDM2, pTEN and TP53 signaling pathway were changed, in which 22 genes were upregulated, while 25 genes were downregulated in PPP2R5C-siRNA799-transfected Jurkat cells. Significantly upregulated genes were SNAI1,APC, CDKN1A, ATM, MDM2 and pTEN, while significantly downregulated gene was identified on GSK-3β. Conclusion PPP2R5C downregulation inhibits Jurkat cells proliferation, which may relate to selective regulation of the genes expression of GSK-3β, MDM2, pTEN and TP53 signaling pathways.

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