Antibody-directed lentiviral vector-mediated hepatocarcinoma cells targeting IGF1R- siRNA/P53 genes therapy

VernacularTitle:肝癌特异性多靶点慢病毒的体外实验研究
Author: Jian NIU ; Hewei ZHAO ; Wen WANG ; Bin YU ; Bin LIU ; Xuehao WANG ; Yewei ZHANG
Publication Type:Journal Article
Keywords: Liver cancer cell; Human insulin like growth factor receptor 1; Wild type P53; Lentivirus
From: Chinese Journal of Hepatobiliary Surgery 2012;18(5):372-376
CountryChina
Language:Chinese
Abstract: ObjectiveTo investigate the targeting infection of single chain antibody againstAFP (scFv anti-AFP) directed lentivirus and the inhibitory effects of a dual-growth inhibition systemon hepatocarcinoma cells.MethodsPlasmids WtP53-pPRIME-miR30-shRNA-IGF1R,pMD2G-Anti-AFP,and psPAX2 have previously been constructed to cotransfect to the packaging cell line 293Tusing Lipofectamine2000.The infection results were observed through fluorescence microscopy.PCRand Western blotting were used to demonstrate the successful transduction and transcription of theWtP53-pPRIME-miR30-shRNA-IGF1R gene.The effects of reconstructed lentivirus infected liver cellgrowth were assessed by the cell growth curve of CCK8 cells. Apoptosis was evaluated by theTUNEL assay.ResultsRecombined lentivirus was successfully constructed with the functional PFUtiters of recombined lentivirus at 4.58× 109PFU/ml.This positive result was confirmed by PCR andWestern blotting.ConclusionsThe targeted therapy mediated by anti-AFP scFv could significantlyinhibit the proliferation of HEP3B cells and promote the apoptosis.