Yang Yu1, Xianzuo Zhang1, Zhongqi Li1, Lei Kong1, Yan Huang2. 1. Department of Orthopedic, The First Affiliated Hospital of the University of Science and Technology of China, Hefei 230001, China. 2. Department of Orthopedic, The First Affiliated Hospital of the University of Science and Technology of China, Hefei 230001, China. Electronic address: huang_USTC@163.com.
Abstract
OBJECTIVE: The aim of this study was to investigate the regulation mechanism of HOTAIR on nucleus pulposus cell (NPC) apoptosis induced by inflammatory cytokines. METHODS: QRT-PCR was performed to analyze the expression of HOTAIR and miR-34a. Safranin O staining and immunohistochemical staining were measured to identify NPCs. ELISA assay was used to detect TNF-α level. Apoptosis was detected with TUNEL assay. Western blotting was measured to analyze the expression of caspase-3, Bax and Bcl-2. TargetScan was used to predict potential targets of miR-34a. RESULTS: HOTAIR was significantly low-expressed in degenerative nucleus pulposus (NP) tissues and cells of IDD patients, overexpressing HOTAIR obviously inhibited TNF-α level, NPCs apoptosis and the expression of caspase-3 and Bax, while promoted the expression of Bcl-2. MiR-34a was obviously expressed in degenerative NP tissues and cells of IDD patients. HOTAIR reduced miR-34a induced apoptosis. Apoptotic inhibition gene Bcl-2 is the target gene of miR-34a, and showed a negative relationship with miR-34a. CONCLUSION: Our data suggest that lncRNA HOTAIR suppresses TNF-α induced NPCs apoptosis by regulating miR-34a/Bcl-2 axis in IDD patients.
OBJECTIVE: The aim of this study was to investigate the regulation mechanism of HOTAIR on nucleus pulposus cell (NPC) apoptosis induced by inflammatory cytokines. METHODS: QRT-PCR was performed to analyze the expression of HOTAIR and miR-34a. Safranin O staining and immunohistochemical staining were measured to identify NPCs. ELISA assay was used to detect TNF-α level. Apoptosis was detected with TUNEL assay. Western blotting was measured to analyze the expression of caspase-3, Bax and Bcl-2. TargetScan was used to predict potential targets of miR-34a. RESULTS:HOTAIR was significantly low-expressed in degenerative nucleus pulposus (NP) tissues and cells of IDDpatients, overexpressing HOTAIR obviously inhibited TNF-α level, NPCs apoptosis and the expression of caspase-3 and Bax, while promoted the expression of Bcl-2. MiR-34a was obviously expressed in degenerative NP tissues and cells of IDDpatients. HOTAIR reduced miR-34a induced apoptosis. Apoptotic inhibition gene Bcl-2 is the target gene of miR-34a, and showed a negative relationship with miR-34a. CONCLUSION: Our data suggest that lncRNA HOTAIR suppresses TNF-α induced NPCs apoptosis by regulating miR-34a/Bcl-2 axis in IDDpatients.
Authors: Kui Huang; Lei Shen; Tieming Niu; Ying Zhao; Jiucun Fu; Yunpeng Cao Journal: Evid Based Complement Alternat Med Date: 2019-02-05 Impact factor: 2.629