条件性肿瘤抑制基因Unc5H4依赖P53诱导人神经母细胞瘤细胞凋亡的实验研究

Conditional tumor supressor Unc5H4 induces apoptosis in neuroblastoma cells depending on P53

目的探讨依赖性受体Unc5H4诱导细胞凋亡功能与P53的关系。方法应用siRNA技术在SHSY5Y细胞中沉默P53基因,应用G418筛选出稳定沉默P53表达的细胞,以时间依赖性模式进行ADR刺激,并检测Unc5H4在mRNA及蛋白水平的表达情况。进一步,在SK-N-BE细胞中转染P53基因表达后,以时间依赖性模式检测Unc5H4在mRNA及蛋白水平的表达情况。结果稳定沉默P53表达的SH-SY5Y细胞,与对照组相比,在ADR刺激下未检测到内源性Unc5H4的表达。在成功表达P53基因的SK-N-BE细胞中,P53基因表达能够在mRNA及蛋白水平诱导Unc5H4的表达。结论 Unc5H4在无配体作用下的促凋亡作用是依赖P53的正常功能完成的,Unc5H4是P53的新型调控基因。

[Objective] To study the relationship between dependence receptor Unc5H4 and P53. [Methods] P53 gene was silent by siRNA method in SH-SY5Y cells, and then the stable clone was selected with G418. ADR was used with time-dependent manner for DNA damage. Unc5H4 expression was measured both in mRNA and protein level. Furthermore, P53 plasmid was transient transfected into SK-N-BE cells. Unc5H4 expression was measured also in both mRNA and protein level with time-dependent manner after P53 expression. [Results] Compared with control, endogenous Unc5H4 expression was not detected in stable clone of P53-silent SH-SY5Y cells after ADR-induced DNA damage. But in SK-N-BE cells, after P53 was successful expressed with transit transfection, Unc5H4 expression was also induced both in mRNA and protein level. [Conclusion] The function of Unc5H4 without ligand to induce apoptosis was dependent on normal P53 function. Unc5H4 was a new kind of regulator gene of P53.