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目的NOR1基因真核表达载体pcDNA3.1(+)的构建并了解其对肝癌细胞系HepG2生长的影响。方法将NOR1cDNA亚克隆至pcDNA3.1(+)真核表达载体上,并把重组质粒pcDNA3.1(+)/NOR1经脂质体转染至HepG2细胞中,运用MTT法、台盼蓝排斥等试验、流式细胞仪等分析NOR1基因对肝癌细胞HepG2生物学活性的影响。结果成功构建了NOR1基因真核表达体pcDNA3.1(+)/NOR1,经pcDNA3.1(+)/NOR1转染后的HepG2细胞生长速度明显抑制(P<0.05),且细胞从G0/G1期进入S期明显延缓。结论重组质粒pcDNA3.1(+)/NOR1能在HepG2细胞内表达,且能影响HepG2细胞的生长。
Objective To construct the eukaryotic expression vector pcDNA3.1 (+) of NOR1 gene and study its effect on the growth of hepatocellular carcinoma cell line HepG2. Methods NOR1 cDNA was subcloned into eukaryotic expression vector pcDNA3.1 (+), and the recombinant plasmid pcDNA3.1 (+) / NOR1 was transfected into HepG2 cells by lipofectamine. MTT assay and trypan blue exclusion Test, flow cytometry analysis of NOR1 gene on HepG2 cells biological activity. Results The eukaryotic expression of NOR1 gene pcDNA3.1 (+) / NOR1 was successfully constructed. The growth of HepG2 cells was inhibited by pcDNA3.1 (+) / NOR1 (P <0.05) S phase into the period was significantly delayed. Conclusion The recombinant plasmid pcDNA3.1 (+) / NOR1 can be expressed in HepG2 cells and can affect the growth of HepG2 cells.