两株NIH-3T3细胞间的基因表达差异研究

Detection of Differentially Expressed Genes in Two Cell Lines of NIH-3T3

通过与细胞膜表面的受体TNFRⅠ结合,肿瘤坏死因子-α(TNF-α)可诱导凋亡或坏死.有两株NIH-3T3细胞在TNF刺激下分别存在两种死亡情况:NIH-3T3(R)为caspase依赖的凋亡;而NIH-3T3(S)的死亡则不依赖caspase.为了深入研究两株细胞在TNF-α诱导下选择不同死亡方式的机理,本研究利用cDNA表达谱芯片对这两株细胞的基因表达差异性进行研究,筛选出差异表达的基因,利用逆转录PCR验证,从而鉴定出两个可能与细胞死亡相关的基因:calpain6和R IP3.

As a proinflammatory cytokine,Tumor Necrosis Factor alpha (TNF-α) plays an important role in many cellular events such as cell proliferation,differentiation and death.TNF-α elicits its biological effects on responding cells by binding to the receptors,TNFRI and TNFRII.But only TNFRI is a death receptor.By binding to the receptor TNFRⅠ,TNF-α induces both apoptotic and necrotic cell death,depending on cell types and treatment condition.The mechanism of TNF-induced non-apoptotic cell death is largely unknown although that of TNF-induced apoptosis has been extensively studied.In this study,we used two lines of NIH-3T3 cells:NIH-3T3 (R) and NIH-3T3 (S). In NIH-3T3 (R) cells,TNF-α induced caspase-dependent cell death;but in NIH-3T3 (S) cells,under a caspase-inhibited condition, TNF-α effectively induced cell death.So we believe that these two cell lines would be good material for studying the difference of the mechanisms between the two distinct cell death pathways induced by TNF-α.Large-scale gene expression profiles between these two cell lines were detected with cDNA microarray.Some of the data were confirmed by RT-PCR.As a result,two genes were identified,which are calpain 6 and RIP3,which may be crucial in mediating TNF-induced cell death.