miR-122表达载体的构建及其对HepG2.2.15细胞增殖的抑制作用

Construction of an miR-122 expression vector and its inhibitory effects on proliferation of HepG2.2.15 cells

目的构建miR-122表达载体并检测其表达对HepG2.2.15细胞恶性表型的逆转作用。方法以HepG2.2.15细胞基因组DNA为模板,PCR扩增miR-122前体cDNA序列,以质粒pSilencer3.1-H1 neo为母本,构建miR-122表达载体将其转染HepG2.2.15细胞。表达后,利用ELISA检测HepG2.2.15细胞HBV复制和表达的变化,利用CCK8检测细胞增殖的变化。结果构建的miR-122表达载体可在HepG2.2.15细胞中表达。转染后HepG2.2.15细胞中HBV复制、表达以及细胞增殖均被抑制。结论 miR-122表达载体可在HepG2.2.15细胞中有效表达,其表达可抑制HBV复制、表达以及细胞增殖。

Objective To construct an miR-122 expression vector and examine the effect of miR-122 expression in reversing the malignant phenotype of the HepG2.2.15 cell line.Methods Genomic DNA of the HepG2.2.15 cell line was used as a template and the target gene fragment was PCR-amplified.The product was confirmed by enzymatic digestion and sequencing.The vector was transfected into HepG2.2.15 cells.HBsAg and HBeAg in the supernatant from cell cultures were measured,and HBV-DNA level was detected by real-time PCR.Cell proliferation after transfection was monitored.Results The MiR-122 expression vector was expressed in HepG2.2.15 cells,leading to inhibition of HBV replication and expression as well as proliferation of HepG2.2.15 cells.Conclusion MiR-122,expressed by means of an expression vector,inhibits HBV replication and expression.Proliferation of HepG2.2.15 cells was also impaired.