目的应用基因芯片进行高通量分析沉默基因BC047440后相关基因表达谱的变化,以了解BC047440通过NF_KB信号途径上游的何种分子调控NF-KB活性。方法应用博奥公司提供的人全基因组基因芯片检测沉默BC047440基因后相关基因表达谱的改变,利用其数据库和MAs分析系统进行分析筛选,寻找NF-kB发生表达改变的上游分子并进行RT—PCR验证。结果沉默BC047440基因后有189个基因表达改变,其中130个基因上调表达2倍以上,59个基因下调表达超过50%,其中NF—kB信号途径上游分子中仅TRAF6下降为对照组的23.06%。RT—PCR验证TRAF6的mRNA的表达下降为对照组的29.5%,与基因芯片结果类似。结论应用基因芯片可以高通量高效率地分析沉默BC047440基因后的基因表达谱,BC047440基因可通过调控上游分子TRAF6作用于NF-KB信号途径来调控肝癌增殖。
Objective To investigate the mechanism that BC047440 gene regulates nuclear factor KB sigal passway and analyze the differential expression gene between HepG2 cells and HepG2 cells BC047440 gene silenced by RNAi using 35K Human Genome Array. Methods The differential expres- sion gene between HepG2 cells and HepG2 cells with BC047440 gene silenced was analyzed by 35K Human Genome Array, and the data were submitted to the database and MAS system of Capitalbio Corporation. Then TRAF6 was confirmed by RT-PCR test. Results Among the total 35000 probe sets, the expression of 59 genes was down-regulated for more than 50~ and 130 genes were up-regulated more than 2 fold in the silencing group when compared with normal controls. TRAF6 mRNA was decreased for 29.5 ~ in silicening HepG2 compared with that of wild HepG2 by RT-PCR, which is similar to human genome array(23.06 %). Conclusiorl The high throughput and effective oligomicroarray can analyze the differential expression gene and BC047440 gene might regulate NF-KB signal pathway inderectly by TRAF6.