Construction of recombinant expression lentivirus vector carrying IL-24 gene and its inhibitory effect on HepG2 growth

Objective To construct the recombinant lentivirus containing interleukin(IL)-24 gene and detect the effect of IL-24 on the growth of human hepatoma cells.Methods Total RNAs were extracted from PBMCs stimulated by 5 μg/ml PHA,IL-24 cDNA amplified by RT-PCR was cloned into the lentivirus vector pHAGE-CMV-MCS-IzsGreen(named as pHAGE-IL-24),and then the recombinant vector pHAGE-IL-24,packaging vector psPAX2 and enveloping vector pMD2.G were cotransfected into the HEK 293T cells.Culture media was harvested and filtered through a 0.45 μm filter to remove the cells.The viral titer was checked by observing the expression of green fluorescent protein(GFP).After infected by the recombinant lentivirus,the mRNA transcription and protein expression of IL-24 in HEK 293T cells were detected by RT-PCR and Western blot,respectively.Then,HepG2 cells were infected by the recombinant lentivirus when MOI was 5.0,and the proliferation of cells was detected by MTT assay.Results The recombinant lentivirus vector carrying IL-24 was constructed successfully.The viral titer was 5×106 TU/ml,and HepG2 cells could be efficiently infected when the MOI was 5.0. In addition,the expression of IL-24 in HepG2 cells infected with lentivirus-IL-24 was detectable.The proliferation of HepG2 cells was inhibited.Conclusion The recombinant lentivirus with high titer and efficient infection of HepG2 cells could be obtained quickly and simply using the lentivirus vectors system,and IL-24 expression could be observed in HepG2 cells infected with lentivirus-IL-24.IL-24 inhibits the proliferation of HepG2 cells.