中文摘要：

目的构建shRNA并干扰肝癌HepG2细胞中BC047440基因，观察其对肝癌HepG2细胞增殖的影响。方法根据BC047440基因序列，设计和合成两条特异表达该基因shRNA序列并克隆载体pGenesil-1，转染肝癌HepG2细胞并用定量PCR观察抑制BC047440基因表达的效果，并通过MTr法和流式细胞仪检测细胞增殖及细胞周期的变化。结果酶切和序列测定提示成功构建了特异表达BC047440基因的shRNA1和shRNA2两个质粒。shRNA1和shRNA2对BC047440mRNA的抑制率分别为80．22％和58．63％。干扰BC047440基因后，肝癌HepG2细胞增殖受到明显抑制，主要停滞在s期。结论构建的shRNA能有效干扰肝癌HepG2细胞BC047440基因的表达，并抑制肝癌HepG2细胞的增殖，提示BC047440基因和肝癌HepG2细胞增殖有关。

英文摘要：

Objective To investigate whether the proliferation of HepG2 cells is influenced by interfering BC047440 gene with small hairpin RNA （shRNA） expression plasmid. Methods According to the sequence of BC047440 gene, 2 pairs of BC047440 gene-specific shRNA （ shRNA1 and shRNA2） were designed and synthesized. After primer annealing, they were inserted into plasmid pGenesil-1 to construct the shRNA expression plasmids. The recombinant plasmids were transfected into HepG2 cells. The expression of BC047440 gene was detected by quantitative fluorescent PCR, the proliferation of HepG2 cells by MTT assay and the changes of cell cycle by flow cytometry. Results Two shRNA expression plasmids were constructed successfully and were confirmed by restriction enzyme digestion and sequencing. Quantitative fluorescent PCR analysis showed that shRNA1 and shRNA2 could specifically inhibit the expression of BC047440 gene in HepG2 cells, with the inhibition rate of 80.22% and 58.63%, respectively. The shRNA effectively inhibited the proliferation of HepG2 cells, and arrested the cell cycle in S phase. Conclusions The shRNA significantly inhibits the expression of BC047440 gene and the proliferation of HepG2 cells. The expression of BC047440 may be correlated with the proliferation of HepG2 cells.