乙醛脱氢酶2基因的可变剪接分析

Alternative splicing analysis of the aldehyde dehydrogenase 2 gene

目的 探讨胃癌细胞中的乙醛脱氢酶2(ALDH2) m RNA发生可变剪接。方法 设计ALDH2基因特异性引物,用胃癌细胞(AGS、HGC-27、MKN45)、正常人胃黏膜上皮细胞GES-1和人外周全血白细胞提取RNA,进行逆转录聚合酶链反应(RT-PCR)扩增,然后对ALDH2扩增产物进行测序分析;使用DNAStar软件分析ALDH2变异体(V)的开放阅读框(ORF)。结果 ALDH2变异体1(V1)外显子2和外显子4发生可变剪接,V1外显子1的3’端和外显子3的5’端3个不同的位点发生可变剪接,V1存在8个长度超过100氨基酸的ORF。结论 ALDH2 m RNA存在多种可变剪接形式,提示其存在复杂的蛋白表达功能。

Objective To investigate the occurrence of alternative splicing of aldehyde dehydrogenase2(ALDH2) mRNA in gastric cancer cells.Methods Specific primers for ALDH2 gene was designed,so as to extract the RNA from gastric cancer cells(AGS,HGC-27,MKN45),normal gastric mucosal epithelial cells(GES-1) and human peripheral blood leukocytes.Then he reverse transcriptase polymerase chain reaction(RT-PCR) was amplified,and the products of ALDH2 amplification were sequenced and analyzed;The open reading frame(ORF) of ALDH2 variant(V) was analyzed using DNAStar software.Results Alternative splicing site occurred in exon 2 and exon 4 of ALDH2 variant1(V1);Alternative splicing occurs at three different sites in the 3’end of V1 exon 1 and the 5’end of exon 3.Eight ORFs with more than 100 amino acids in length existed in V1.Conclusion In ALDH2 m RNA there exist multiple alternative splicing forms,suggesting that it has complex functions of protein expression.