Song-Shu Lin1,2,3, Chi-Chien Niu1,3,4, Li-Jen Yuan5, Tsung-Ting Tsai1,3, Po-Liang Lai1,3, Kowit-Yu Chong3,6, Kuo-Chen Wei3,7, Chiung-Yin Huang3,6, Meng-Ling Lu8, Chuen-Yung Yang1,3, Steve W N Ueng9,10,11. 1. Department of Orthopaedic Surgery, Chang Gung Memorial Hospital, No 5, Fu-Hsing Street, Linkou, Taoyuan, 333, Taiwan. 2. Department of Nursing, Chang Gung University of Science and Technology, Taoyuan, Taiwan. 3. Hyperbaric Oxygen Medical Research Lab, Bone and Joint Research Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan. 4. College of Medicine, Chang Gung University, Taoyuan, Taiwan. 5. Department of Orthopaedic Surgery, E-Da Hospital/I-Shou University, Kaohsiung, Taiwan. 6. Department of Medical Biotechnology and Laboratory Science, College of Medicine, Chang Gung University, Taoyuan, 333, Taiwan. 7. Department of Neurosugery, New Taipei Municipal Tu Cheng Hospital, Chang Gung Memorial Hospital and Chang Gung University, Taoyuan, Taiwan. 8. Department of Orthopaedic Surgery, Chang Gung Memorial Hospital, Kaohsiung, Taiwan. 9. Department of Orthopaedic Surgery, Chang Gung Memorial Hospital, No 5, Fu-Hsing Street, Linkou, Taoyuan, 333, Taiwan. wenneng@adm.cgmh.org.tw. 10. Hyperbaric Oxygen Medical Research Lab, Bone and Joint Research Center, Chang Gung Memorial Hospital, Taoyuan, Taiwan. wenneng@adm.cgmh.org.tw. 11. College of Medicine, Chang Gung University, Taoyuan, Taiwan. wenneng@adm.cgmh.org.tw.
Abstract
BACKGROUND: MicroRNA (miRNA) plays a vital role in the intervertebral disc (IVD) degeneration. The expression level of miR-573 was downregulated whereas Bax was upregulated notably in human degenerative nucleus pulposus cells. In this study, we aimed to investigate the role of miR-573 in human degenerative nucleus pulposus (NP) cells following hyperbaric oxygen (HBO) treatment. METHODS: NP cells were separated from human degenerated IVD tissues. The control cells were maintained in 5% CO2/95% air and the hyperoxic cells were exposed to 100% O2 at 2.5 atmospheres absolute. MiRNA expression profiling was performed via microarray and confirmed by real-time PCR, and miRNA target genes were identified using bioinformatics and luciferase reporter assays. The mRNA and protein levels of Bax were measured. The proliferation of NPCs was detected using MTT assay. The protein expression levels of Bax, cleaved caspase 9, cleaved caspase 3, pro-caspase 9, and pro-caspase 3 were examined. RESULTS: Bioinformatics analysis indicated that the 3' untranslated region (UTR) of the Bax mRNA contained the "seed-matched-sequence" for hsa-miR-573, which was validated via reporter assays. MiR-573 was induced by HBO and simultaneous suppression of Bax was observed in NP cells. Knockdown of miR-573 resulted in upregulation of Bax expression in HBO-treated cells. In addition, overexpression of miR-573 by HBO increased cell proliferation and coupled with inhibition of cell apoptosis. The cleavage of pro-caspase 9 and pro-caspase 3 was suppressed while the levels of cleaved caspase 9 and caspase 3 were decreased in HBO-treated cells. Transfection with anti-miR-573 partly suppressed the effects of HBO. CONCLUSION: Mir-573 regulates cell proliferation and apoptosis by targeting Bax in human degenerative NP cells following HBO treatment.
BACKGROUND: MicroRNA (miRNA) plays a vital role in the intervertebral disc (IVD) degeneration. The expression level of miR-573 was downregulated whereas Bax was upregulated notably in human degenerative nucleus pulposus cells. In this study, we aimed to investigate the role of miR-573 in human degenerative nucleus pulposus (NP) cells following hyperbaric oxygen (HBO) treatment. METHODS: NP cells were separated from human degenerated IVD tissues. The control cells were maintained in 5% CO2/95% air and the hyperoxic cells were exposed to 100% O2 at 2.5 atmospheres absolute. MiRNA expression profiling was performed via microarray and confirmed by real-time PCR, and miRNA target genes were identified using bioinformatics and luciferase reporter assays. The mRNA and protein levels of Bax were measured. The proliferation of NPCs was detected using MTT assay. The protein expression levels of Bax, cleaved caspase 9, cleaved caspase 3, pro-caspase 9, and pro-caspase 3 were examined. RESULTS: Bioinformatics analysis indicated that the 3' untranslated region (UTR) of the Bax mRNA contained the "seed-matched-sequence" for hsa-miR-573, which was validated via reporter assays. MiR-573 was induced by HBO and simultaneous suppression of Bax was observed in NP cells. Knockdown of miR-573 resulted in upregulation of Bax expression in HBO-treated cells. In addition, overexpression of miR-573 by HBO increased cell proliferation and coupled with inhibition of cell apoptosis. The cleavage of pro-caspase 9 and pro-caspase 3 was suppressed while the levels of cleaved caspase 9 and caspase 3 were decreased in HBO-treated cells. Transfection with anti-miR-573 partly suppressed the effects of HBO. CONCLUSION:Mir-573 regulates cell proliferation and apoptosis by targeting Bax in human degenerative NP cells following HBO treatment.
Authors: S A Susin; H K Lorenzo; N Zamzami; I Marzo; B E Snow; G M Brothers; J Mangion; E Jacotot; P Costantini; M Loeffler; N Larochette; D R Goodlett; R Aebersold; D P Siderovski; J M Penninger; G Kroemer Journal: Nature Date: 1999-02-04 Impact factor: 49.962
Authors: Rajesha Rupaimoole; Sherry Y Wu; Sunila Pradeep; Cristina Ivan; Chad V Pecot; Kshipra M Gharpure; Archana S Nagaraja; Guillermo N Armaiz-Pena; Michael McGuire; Behrouz Zand; Heather J Dalton; Justyna Filant; Justin Bottsford Miller; Chunhua Lu; Nouara C Sadaoui; Lingegowda S Mangala; Morgan Taylor; Twan van den Beucken; Elizabeth Koch; Cristian Rodriguez-Aguayo; Li Huang; Menashe Bar-Eli; Bradly G Wouters; Milan Radovich; Mircea Ivan; George A Calin; Wei Zhang; Gabriel Lopez-Berestein; Anil K Sood Journal: Nat Commun Date: 2014-10-29 Impact factor: 14.919