肝癌相关基因MEF2D反义RNA的鉴定及分析

Identification and analysis of hepatocellular carcinoma associated antisense RNA MEF2D-AS

鉴定和分析肝癌细胞中新的天然反义RNA MEF2D-AS根据已建立的双链RNA文库筛选与肝癌相关的反义RNA.通过RT-qPCR验证其反义RNA的存在,采用cDNA末端快速扩增技术获得其全长序列,并检测HepG2细胞经5-Aza-dC处理前后MEF2D基因正反义链表达量的变化.结果显示,成功鉴定出1条全长为1 265bp的新的天然反义RNA,并预测其属于长链非编码RNA(long non-coding RNA,简称为lncRNA),命名为MEF2D-AS.经5-Aza-dC处理后,MEF2D-AS的表达量升高,而MEF2DmRNA的表达量降低.证实了MEF2D-AS的存在并预测其属于长链非编码RNA,且与MEF2D正反义RNA是反相关关系.此研究结果可为进一步探讨肝癌发生机制提供参考.

To identify and analyze new natural antisense RNAs in hepatoma cells, HCC antisense RNA was screened based on established double-stranded RNA library. RT-qPCR was used to detect the expression of the MEF2D gene and MEF2D-AS gene before and after 5 Aza dC treatment. A new natural antisense RNA was successfully identified, which was 1 265 bp in length. It was predicted to belong to a long non-coding RNA （IneRNA） and named MEF2D-AS. After treatment with 5-Aza-dC, the expression of MEF2D-AS increased, while the expression of MEF2D mRNA decreased. MEF2D-AS has been confirmed to be a long non-coding RNA. There is an inverse correlation between MEF2D and MEF2D-AS expression. This discovery is of significance for exploring mechanism of HCC tumorigenesis.