title: MicroRNA-302 induces proliferation and inhibits oxidant-induced cell death in human adipose tissue-derived mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) are a heterogeneous population of cells that proliferate in vitro as plastic-adherent cells, have a fibroblast-like morphology, form colonies in vitro and can differentiate into bone, cartilage and fat cells. The abundance, ease and repeatable access to subcutaneous adipose tissue and the simple isolation procedures provide clear advantages for the use of human adipose tissue-derived mesenchymal stem cells (hASDCs) in clinical applications. We screened microRNAs (miRNAs) that affected the proliferation and survival of hADSCs. Transfection of miR-302d mimic increased cell proliferation and protected cells from oxidant-induced cell death in hADSCs, which was supported by flow-cytometric analysis. miR-302d did not affect the expression of Bcl-2 family members or anti-oxidant molecules. The Nrf2-Keap1 system, which is one of the major mechanisms for the cellular defense against oxidative stress, was not altered by transfection of miR-302d mimic. To identify the target of the miR-302d actions on proliferation and survival of hADSCs, a microarray analysis was performed using miR-302d-overexpressing hADSCs. Real-time PCR analysis showed that transfection of miR-302d mimic inhibited the CDKN1A and CCL5 expression. Downregulation of CDKN1A with a specific siRNA mimicked the effect of miR-302d on hADSCs proliferation, but did not affect miR-302d-induced cell survival. Downregulation of CCL5 protected oxidant-induced cell death as miR-302d, inhibited oxidant-induced reactive oxygen species (ROS) generation and the addition of recombinant CCL5 inhibited the protective action of miR-302d on oxidant-induced cell death. This study indicates that miR-302 controls proliferation and cell survival of hADSCs through different targets and that this miRNA can be used to enhance the therapeutic efficacy of hADSCs transplantation in vivo.
authors.affiliation: 1] Department of Physiology, School of Medicine, Pusan National University, Yangsan 626-870, Korea [2] Medical Research Center for Ischemic Tissue Engineering, Pusan National University, Yangsan 626-870, Korea [3] BK21 Medical Science Education Center, School of Medicine, Pusan National University, Yangsan 626-870, Korea.
authors.forename: J Y
authors.initials: JY
authors.lastname: Kim
authors.affiliation: 1] Department of Physiology, School of Medicine, Pusan National University, Yangsan 626-870, Korea [2] Medical Research Center for Ischemic Tissue Engineering, Pusan National University, Yangsan 626-870, Korea [3] BK21 Medical Science Education Center, School of Medicine, Pusan National University, Yangsan 626-870, Korea.
authors.forename: K K
authors.initials: KK
authors.lastname: Shin
authors.affiliation: 1] Department of Physiology, School of Medicine, Pusan National University, Yangsan 626-870, Korea [2] Medical Research Center for Ischemic Tissue Engineering, Pusan National University, Yangsan 626-870, Korea [3] BK21 Medical Science Education Center, School of Medicine, Pusan National University, Yangsan 626-870, Korea.
authors.forename: A L
authors.initials: AL
authors.lastname: Lee
authors.affiliation: 1] Department of Physiology, School of Medicine, Pusan National University, Yangsan 626-870, Korea [2] Medical Research Center for Ischemic Tissue Engineering, Pusan National University, Yangsan 626-870, Korea [3] BK21 Medical Science Education Center, School of Medicine, Pusan National University, Yangsan 626-870, Korea.
authors.forename: Y S
authors.initials: YS
authors.lastname: Kim
authors.affiliation: 1] Department of Physiology, School of Medicine, Pusan National University, Yangsan 626-870, Korea [2] Medical Research Center for Ischemic Tissue Engineering, Pusan National University, Yangsan 626-870, Korea [3] BK21 Medical Science Education Center, School of Medicine, Pusan National University, Yangsan 626-870, Korea.
authors.forename: H J
authors.initials: HJ
authors.lastname: Park
authors.affiliation: 1] Department of Physiology, School of Medicine, Pusan National University, Yangsan 626-870, Korea [2] Medical Research Center for Ischemic Tissue Engineering, Pusan National University, Yangsan 626-870, Korea.
authors.forename: Y K
authors.initials: YK
authors.lastname: Park
authors.affiliation: Department of Plastic Surgery, School of Medicine, Pusan National University, Pusan 602-739, Korea.
authors.forename: Y C
authors.initials: YC
authors.lastname: Bae
authors.affiliation: 1] Department of Physiology, School of Medicine, Pusan National University, Yangsan 626-870, Korea [2] Medical Research Center for Ischemic Tissue Engineering, Pusan National University, Yangsan 626-870, Korea [3] BK21 Medical Science Education Center, School of Medicine, Pusan National University, Yangsan 626-870, Korea [4] Medical Research Institute, Pusan National University, Pusan 602-739, Korea.
authors.forename: J S
authors.initials: JS
authors.lastname: Jung
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D017209:Apoptosis / Q000187:drug effects; D001483:Base Sequence; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D018946:Chemokine CCL5 / Q000378:metabolism; D003035:Cobalt / Q000633:toxicity; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015536:Down-Regulation / Q000187:drug effects; D059585:G1 Phase Cell Cycle Checkpoints; D051547:Heme Oxygenase-1 / Q000235:genetics / Q000378:metabolism; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism; D000072019:Kelch-Like ECH-Associated Protein 1; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism*; D035683:MicroRNAs / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008981:Molsidomine / Q000031:analogs & derivatives / Q000633:toxicity; D051267:NF-E2-Related Factor 2 / Q000235:genetics / Q000378:metabolism; D016376:Oligonucleotides, Antisense / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D016415:Sequence Alignment; D059467:Transcriptome
chemical_list: D018946:Chemokine CCL5; D050759:Cyclin-Dependent Kinase Inhibitor p21; D047908:Intracellular Signaling Peptides and Proteins; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D035683:MicroRNAs; D051267:NF-E2-Related Factor 2; C495635:NFE2L2 protein, human; D016376:Oligonucleotides, Antisense; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D003035:Cobalt; C002385:linsidomine; D008981:Molsidomine; C496541:HMOX1 protein, human; D051547:Heme Oxygenase-1; C018021:cobaltous chloride