中文摘要：

目的：建立甲胎蛋白（AFP）表达缺失的HepG2细胞株。方法：选择AFP的RNA干涉靶序列，体外合成两段互补的寡核苷酸，与线性化的pll3．7连接，扩增纯化得到所需质粒。鉴定后将质粒用慢病毒包装、纯化得到病毒粗提液，与HepG2细胞共培养，转染后用RT—PCR和Westernblot实验检测细胞AFP的表达情况。结果：RT-PCR和Westernblot实验显示，干扰后HepG2细胞AFP基因和蛋白的表达显著减少。干扰效率达84％。所得细胞株传代后仍仅能检测到微量AFP表达。结论：成功构建稳定的AFP表达接近缺失的HepG2模型细胞株。

英文摘要：

Objective： To construct the model cell line which was out of expression of the AFP of HepG2 cell line. Methods： Two complementary oligonucleotides were synthesized in vitro, which was target AFP, then connected the linearization pl13.7, amplificated and purified plasmid. The plasmid for AFP was transfected into HepG2 cells by lentivirus. Real Time PCR and Western blot assay were applied to detect the expression of AFP. Results： After infection, the expression of AFP in HepG2 cells was down-regulated （P 〈 0. 01 ） . The interference rate was over 84%. After subculturing, cells were also loss expression of AFP. Conclusion： The model cell line loss expression of AFP is established.