title: Cell-specific expression of artificial microRNAs targeting essential genes exhibit potent antitumor effect on hepatocellular carcinoma cells.
abstract: To achieve specific and potent antitumor effect of hepatocyte carcinoma cells, replication defective adenoviral vectors, namely rAd/AFP-amiRG, rAd/AFP-amiRE and rAd/AFP-amiRP, were constructed which were armed with artificial microRNAs (amiRs) targeting essential functional genes glyceraldehyde-3-phosphate dehydrogenase, eukaryotic translation initiation factor 4E and DNA polymerase α respectively under the control of a recombinant promoter comprised of human α-fetoprotein enhancer and basal promoter. The AFP enhancer/promoter showed specific high transcription activity in AFP-positive HCC cells Hep3B, HepG2 and SMMC7721, while low in AFP-negative cell Bcap37. All artificial microRNAs exhibited efficient knockdown of target genes. Decreased ATP production and protein synthesis was observed in rAd/AFP-amiRG and rAd/AFP-amiRE treated HCC cells. All three recombinant adenoviruses showed efficient blockage of cell cycle progression and significant suppression of HCC cells in vitro. In nude mice model bearing Hep3B xenograft, administration of rAd/AFP-amiRG showed potent antitumor effect. The strategy of tumor-specific knockdown of genes essential for cell survival and proliferation may suggest a novel promising approach for HCC gene therapy.
authors.affiliation: Clinical Research Center, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, P. R. China.|Cancer Center, The First Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, P. R. China.
authors.forename: Chenyu
authors.initials: C
authors.lastname: Mao
authors.affiliation: Institute of Occupational Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang, P. R. China.
authors.forename: Hao
authors.initials: H
authors.lastname: Liu
authors.affiliation: Sir Run Run Shaw Institute of Clinical Medicine, Zhejiang University, Hangzhou, Zhejiang, P. R. China.
authors.forename: Ping
authors.initials: P
authors.lastname: Chen
authors.affiliation: Clinical Research Center, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, P. R. China.
authors.forename: Jingjia
authors.initials: J
authors.lastname: Ye
authors.affiliation: Cancer Center, The First Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, P. R. China.
authors.forename: Lisong
authors.initials: L
authors.lastname: Teng
authors.affiliation: Institute of Occupational Diseases, Zhejiang Academy of Medical Sciences, Hangzhou, Zhejiang, P. R. China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Jia
authors.affiliation: Clinical Research Center, The Second Affiliated Hospital of Zhejiang University School of Medicine, Hangzhou, Zhejiang, P. R. China.
authors.forename: Jiang
authors.initials: J
authors.lastname: Cao
mesh_terms: D000818:Animals; D006528:Carcinoma, Hepatocellular / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000235:genetics; D055785:Gene Knockdown Techniques / Q000379:methods*; D020043:Genes, Essential; D015316:Genetic Therapy / Q000379:methods*; D056945:Hep G2 Cells; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology / Q000628:therapy*; D051379:Mice; D008807:Mice, Inbred BALB C; D035683:MicroRNAs / Q000008:administration & dosage* / Q000096:biosynthesis / Q000235:genetics; D000091402:Prohibitins; D011401:Promoter Regions, Genetic; D034622:RNA Interference; D011897:Random Allocation; D014158:Transcription, Genetic; D014162:Transfection; D023041:Xenograft Model Antitumor Assays; D000509:alpha-Fetoproteins / Q000235:genetics
chemical_list: C097353:AFP protein, human; D035683:MicroRNAs; C000717331:PHB2 protein, human; C000717333:Phb2 protein, mouse; D000091402:Prohibitins; D000509:alpha-Fetoproteins
keywords: RNA interference; artificial microRNA; gene therapy; hepatocelluar carcinoma