title: Lentiviral vector-mediated RNA interference targeted against prohibitin             inhibits apoptosis of the retinoic acid-resistant acute promyelocytic leukemia             cell line NB4-R1.
abstract: To investigate the possibility of prohibitin (PHB) inhibition by lentiviral vector-mediated RNA interference (RNAi) and its influence on cell apoptosis in the retinoic acid-resistant acute promyelocytic leukemia cell line NB4-R1, a lentiviral vector encoding a short hairpin RNA (shRNA) targeted against PHB (pGCSIL-GFP-PHB) was constructed and transfected into the packaging cells 293T, and the viral supernatant was collected to transfect NB4-R1 cells. Quantitative real-time fluorescent PCR and western blotting were used to detect the expression levels of PHB. Flow cytometry and detection of enzymatic activity of caspase-3 by western blotting were employed to examine cell apoptosis. Our results provide evidence that the lentiviral vector pGCSIL-GFP-PHB was constructed successfully, and the PHB mRNA and the protein expression inhibitory rates were 90.3 and 95.8%, respectively. When compared to the control group, the activity of caspase-3 decreased significantly, which showed a 57.3% downregulation, and the apoptosis rate was reduced by 44.6% (P<0.05). In conclusion, downregulation of the PHB gene may inhibit apoptosis of NB4-R1 cells, and it is speculated that this was at least partly due to the downregulation of caspase-3, and PHB may be a novel target for gene therapy for retinoic acid-resistant acute promyelocytic leukemia.
authors.affiliation: Department of Hematology, the First Affiliated Hospital, School                     of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Liu
authors.forename: Pengcheng
authors.initials: P
authors.lastname: He
authors.forename: Mei
authors.initials: M
authors.lastname: Zhang
authors.forename: Di
authors.initials: D
authors.lastname: Wu
mesh_terms: D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D045744:Cell Line, Tumor; D015536:Down-Regulation; D019008:Drug Resistance, Neoplasm / Q000187:drug effects; D005822:Genetic Vectors / Q000378:metabolism*; D057809:HEK293 Cells; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism / Q000473:pathology; D000091402:Prohibitins; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D012097:Repressor Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: C000717328:PHB protein, human; D000091402:Prohibitins; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D012097:Repressor Proteins; D014212:Tretinoin; D053148:Caspase 3