中文摘要：

目的构建人GW112基因的逆转录病毒载体，并通过病毒介导其在人胃癌SGC-7901细胞的稳定表达。方法采用DNA重组技术构建并鉴定重组逆转录病毒载体pLEGFP—N1-GW112；以脂质体PT67细胞包装病毒，经病毒感染的SGC-7901细胞通过G418筛选及荧光定量PCR鉴定，建立GW112稳定过表达SGC-7901细胞株。结果成功构建了pLEGFP-N1-GW112逆转录病毒表达载体，经病毒包装的重组转录病毒成功感染SGC-7901细胞，Real TimePCR结果显示实验组中GW112基因是对照组细胞中的4．2倍。结论逆转录病毒介导的GW112基因在胃癌SGC-7901细胞获得了稳定过表达，为进一步研究GW112在胃癌中的功能奠定了良好基础。

英文摘要：

Objective To construct pLEGFP-N1-GW112 retrovirus expression vector and increase the expression of human GWll2 gene in human gastric cancer SGC-7901 cells with retrovirus-mediated method. Methods The retrovirus vector pLEGFP- N1-GWll2 containing signal peptide sequence was constructed by DNA recombinant technology and then transfected into Retro- Packed PT67 cell line with Lipofectamine 2000. The viral supernatants were obtained from the PT67 clones and used to infect SGC- 7901 cells. SGC-7901-GW112 and control SGC-7901-GFP cells were established after selected by G418. Real Time-PCR were used to analyze GW112 mRNA expression. Results The recombinant retrovirus vector pLEGFP-N1-GW112 containing signal peptide was successfully constructed,meanwhile virus package cell line were established and the high titer recombined retrovirus virus were obtained from PT67 clones and infected SGC-7901 cell successfully. Real-Time PCR results revealed GWll2 mRNA levels were increased 4.2 fold in SGC-7901-GWl12 cell compaired with its control. Conclusion SGC-7901-GW112 cells stablly overexpressing GW112 and SGC-7901-GFP were established,which may provide good basic for functional research of GW112 in gastric cancer.