乙肝相关性肝癌差异表达基因的获得及FOLR1分析

Isolation of FOLR1 gene related to hepatocellular carcinoma

运用基因芯片技术获取以稳定转染HBx基因的肝癌细胞HepG2(HepG2-X)及非转染的肝癌细胞HepG2中差异表达的基因,并对其中一条基因进行的生物信息学分析。采用人肝癌G2细胞(HepG2)细胞系为对照组,以稳定转染HBx的HepG2细胞为实验组,抽取总RNA,经过反转录cDNA,对照组用Cy3实验组用Cy5荧光标记,获得cDNA探针;经杂交、洗涤后,通过ImaGene3.0软件进行分析统计。通过基因芯片筛选,获得643条与乙肝相关性肝细胞性癌相关的基因,其中FOLR1基因差异性表达最显著,HBx显著下调其表达,同源性比较分析结果表明,其碱基序列与已经报道的其他12种哺乳动物的相似率为67%-99%,且符合种属之间的进化关系。基因芯片筛选HBx诱导的HepG2差异表达基因具有样品用量少,高质量,高速度,高敏感等特性。FOLR1可能为HBV相关肝癌的发生、转移的诊断、靶基因治疗和预后评估提供一定的依据。

To obtain differentially expressed genes related to human HepG2 using cDNA microarray and evaluate the characterization of FOLR1. Total RNA was extracted and rnRNA was used to make probes, After hybridization and washing procedure ,the ruselts of hybriziation were saenned using ImaGene3.0 software. 643 differentially expressed genes to HepG2 were obtained . FOLR1 was low -expressed in Experimental group and overexpressed in control group, The analysis of BLASTp showed that the isolated FOLR1 shows 67%- 99% identity with the others species. eDNA microarray technology can be successfully applied to identify differentially expressed genes, the FOLR1 gene of human may be correlate with pathogenesis of human HCC. HBx may play an important role in angiogenesis of hepatocellular carcinoma by downregulating FOLRlexpression.