FAM3B基因cDNA克隆及其在胃癌细胞系BGC-823的表达

目的克隆FAM3B cDNA,构建FAM3B重组表达载体并在胃癌细胞系BGC-823中表达,观察其对细胞系的影响,探索新的肿瘤治疗途径。方法构建pEGFP-N2-FAM3B真核表达质粒,将pEGFP-N2-FAM3B转染人胃癌细胞系BGC-823,研究其对胃癌细胞产生的作用。结果重组pEGFP-N2-FAM3B真核表达质粒构建成功,经pEGFP-N2-FAM3B转染后,荧光显微镜下可见转染的BGC-823细胞有绿色荧光蛋白的表达,转染后的BGC-823细胞生长减慢。结论构建完成真核表达载体pEGFP-N2-FAM3B,重组pEGFP-N2-FAM3B质粒在胃癌细胞系BGC-823细胞内成功表达。

Cloning and expression of FAM3B gene in gastric carcinoma cell line BGC-823

Objective To construct an eukaryotic expression vector of human FAM3B full-length gene, pEGFP-N2-FAM3B, transfect it into gastric cancer cell line BGC-823 and observe its effects on BGC-823, in a search for new approach of tumor therapy. Method The full-length gene sequence of FAM3B (708 bp) was obtained with RT-PCR using human gastric epithelium mRNA as the template. The PCR products were transferred into pEGFP-N2 vector using gene recombinant techniques. The plasmid of interest was detected by restrictive enzyme digestion and PCR, and then transfected into BGC-823 cells by lipid reagent. Results The pEGFP-N2-FAM3B eukaryotic expression vector was constructed successfully. The expression of green fluorescence protein (GFP) was observed in the human gastric cancer cell line BGC-823 after pEGFP-FAM3B transfection under fluorescence microscopy. Conclusion The eukaryotic expression vector pEGFP-N2-FAM3B was successfully constructed and expressed in BGC-823 cells.