title: Silencing of the human SET gene in vitro with lentivirus-mediated RNA             interference.
abstract: In our previous study, SET was identified as one of the differentially expressed             proteins that was associated with tetra-arsenic tetra-sulfide (As4S4)-induced             NB4-R1 [retinoic acid-resistant acute promyelocytic leukemia (APL) cell line]             apoptosis. However, the mechanism through which SET regulates pathways during             this process remains unclear. The aim of this study was to construct lentivirus-mediated             short hairpin RNA (shRNA) against SET and investigate the effect of SET on As4S4-induced             retinoic acid-resistant APL cell apoptosis. In the present study, 4 different             oligonucleotides targeting the human SET gene were synthesized and cloned into             the eukaryotic expression plasmid pGCSIL-GFP. The recombinant vectors were introduced             into NB4-R1 cells. The silencing efficiency was measured by real-time quantitative             PCR (RT-qPCR) and western blotting. Our results showed that the 4 recombinant             RNA interference (RNAi) vectors were constructed successfully. Fluorescence microscopy             demonstrated that infection efficiency ranged from 70 to 90%. Infection with the             4 different RNAi vectors significantly knocked down the expression of SET by 52.8,             69.1, 48.9 and 90.3% at the mRNA level, and 92.5, 96.3, 91.7 and 98.4% at the             protein level, respectively. We attempt to clarify the mechanism of As4S4 treatment             on retinoic acid-resistant APL.
authors: Department of Hematology, The First Affiliated Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an 710061, PR China.
authors: Yanfeng
authors: Y
authors: Liu
authors: Pengcheng
authors: P
authors: He
authors: Mei
authors: M
authors: Zhang
authors: Lili
authors: L
authors: Shi
authors: Huachao
authors: H
authors: Zhu
authors: Yuan
authors: Y
authors: Wang
authors: Jing
authors: J
authors: Zhao
mesh_terms: D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D056488:Histone Chaperones / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D015473:Leukemia, Promyelocytic, Acute / Q000378:metabolism / Q000473:pathology; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D034741:RNA, Small Interfering / Q000378:metabolism; D014157:Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D004268:DNA-Binding Proteins; D056488:Histone Chaperones; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C513473:SET protein, human; D014157:Transcription Factors; D014212:Tretinoin