中文摘要：

目的 建立起能够稳定表达NLRC5基因的肝癌HepG2细胞株。方法 设计遗传霉素（G418）的浓度梯度,通过对HepG2细胞筛选最终确定筛选药物浓度。将构建好的人源pEGFP-C2-NLRC5重组质粒利用脂质体介导法转染至HepG2肝癌细胞株中,用G418筛选出耐药阳性克隆。通过免疫荧光技术观察绿色荧光蛋白稳定表达情况,Western blot法验证NLRC5蛋白表达情况。结果 G418在14d内使HepG2细胞全部死亡的最小浓度是300ng/ml。用G418筛选瞬转pEGFP-C2-NLRC514d在荧光显微镜下可见耐药克隆的形成。Westernblot法检测显示,稳定过表达组NL-RC5的蛋白水平比空质粒转染组高（t=15.356,P〈0.05）。结论 成功构建稳定表达NLRC5基因的肝癌细胞株,为进一步研究NLRC5基因在肝癌的体内实验奠定了基础。

英文摘要：

Objective To establish a human hepatoma cell line HepG2 with over-expression of NLRCS. Methods Identified the suitable G418 concentration in the selection of stable transfection HepG2 cell. The eukaryotic ex- pression plasmid pEGFP-C2-NLRC5 was transfected into the HepG2 cells mediated by lipofectamine and then se- lected with G418. Immunofluorescence assay and Western blot were used to analyze the expression of NLRC5. Re- sults HepG2 cells were killed by G418 after 14 days with the minimum concentration of 300 ng/pd. Drug-resistant clones were formed after 14 days by selecting with G418. In the protein level, pEGFP-C2-NLRC5 HepG2 cell line was higher expression than pEGFP-C2 HepG2 cell line, the difference was statistically significant. Conclusion A hepatoma stable cell line over-expressing NLRC5 is successfully established, which may provide critical foundation for functional research of NLRC5 in liver cancer.