中文摘要：

目的构建稳定表达miR-218的AGS细胞系。方法合成miR-218的前体cDNA，并将其插入到pRNAi—CMV3．2miR真核表达载体上。用脂质体Lipofectamine2000介导重组质粒导入AGS细胞。用含G418的培养基筛选稳定表达miR-218的细胞株，并用TaqMan定量PCR方法鉴定稳转细胞株的稳定性。结果分离培养出稳定高表达miR-218的单克隆细胞株。结论该实验成功建立了稳定表达miR-218的AGS细胞系，从而为今后研究miR-218的功能提供实验基础。

英文摘要：

Objective To construct the expression vectors of miR-218 and transfect to AGS cells in order to establish the stably transfected AGS cell lines. Methods The cDNAs of pre-miR-218-1 and pre-miR-218-2 were cloned into pRNAi-CMV3.2 eukaryotic expression vector, respectively, yielding miR-218-1 -pRNAi-CMV3.2 and miR-218-2-pRNAi-CMV3.2. The miR - 218 - 1 -pRNAi-CMV3.2 and miR-218-2-pRNAi-CMV3.2 were transfected into AGS cells by Lipofectamine 2000, respectively： TaqMan quantitative real-time polymerase chain reaction was used to confirm the stableness of transfected AGS after G418 selection. Results Stably transfected AGS cell lines were established. Conclusion The study successfully established the stably transfected AGS cell lines of mir-218 ,and provided the experimental basis for function of miR-218 in the future.