Retrovirus-mediated ARLTS1 gene expression suppresses HepG2 cells proliferation

【Objective】To construct the retroviral vector expressing ARLTS1 gene(PLEGFP-IRES-ARLTS1) and to investigate the effect of exogenous ARLTS1 on proliferation in hepatoma carcinoma cell HepG2.【Methods】 ARLTS1 cDNA was amplified by RT-PCR and subcloned into recombinant retrovirus vector PLEGFP-IRES.Plasmids were transfected into retrovirus packaging cell PT67.The target cells HepG2 were infected with retroviral particles,fluorescence microscopy detected expression of GFP in HepG2 cells.The expression of ARLTS1 protein was determined by Western Blot.The proliferation of cells was analysised by MTT assay.【Results】Restrictive endonuclease identification and gene sequencing confirmed that the recombinant vector was successfully constructed.Fluorescence microscopy observed the expression of GFP in target cells.Western Blot demonstrated the level of ARLTS1 protein was obviously increased in HepG2-ARLTS1 cells.The proliferation rate of HepG2-ARLTS1 cells group was lower than control groups(P 0.05).【Conclusions】The HepG2 cells line for stable expressing ARLTS1 was established and expression of exogenous ARLTS1 decreased HepG2 cells viability.These results laid a foundation for further research the anti-tumor mechanism of ARLTS1.