Knockdown of Long Non-Coding RNA RP11-445H22.4 Alleviates LPS-Induced Injuries by Regulation of MiR-301a in Osteoarthritis

Background/Aims: Several long non-coding RNAs (lncRNAs) play vital roles in osteoarthritis (OA), whereas the role of lncRNA RP11-445H22.4 in OA remains unclear. The study aimed to investigate the effect of lncRNA RP11-445H22.4 on lipopolysaccharide (LPS)-induced cell viability, apoptosis and inflammatory injury of OA. Methods: The expression of RP11-445H22.4, miR-301a and CXCR4 in human cartilage ATDC5 cells were altered by transfection, and then cells were exposed to 5 µg/ml LPS for 12 h. Then cell viability, apoptosis, apoptosis-related factors and inflammatory cytokines were analyzed by CCK-8, flow cytometry, western blot, RT-qPCR and ELISA, respectively. Dual-luciferase reporter assay was performed to assess the binging sites of RP11-445H22.4 and miR-301a. The signal pathways of NF-κB and MAPK/ ERK were determined by western blot. Results: LPS reduced cell viability, increased apoptosis and stimulated release of IL-1β, IL-6, IL-8 and TNF-α. However, RP11-445H22.4 inhibition significantly rescued LPS-induced injuries by promoting cell viability, suppressing apoptosis and inflammatory cytokines secretions in ATDC5 cells. In addition, miR-301a directly bound to RP11-445H22.4, and suppression of miR-301a inversed the effects of RP11-445H22.4 inhibition. Furthermore, CXCR4 was a direct target of miR-301a, and CXCR4 silencing increased cell viability, decreased apoptosis and inflammatory cytokines secretions in LPS-treated ATDC5 cells. Besides, we found that CXCR4 silencing blocked LPS-activated NF-κB and MAPK/ERK pathways. Conclusions: The study indicated that lncRNA RP11-445H22.4-miR-301a-CXCR4 axis played an important role in cartilage ATDC5 cells and provided a theoretical basis of lncRNA RP11-445H22.4 in OA.

[1]  F. Zhao,et al.  Downregulation of miR-221-3p contributes to IL-1β-induced cartilage degradation by directly targeting the SDF1/CXCR4 signaling pathway , 2017, Journal of Molecular Medicine.

[2]  Z. yin,et al.  Inhibition of the p38‐MAPK signaling pathway suppresses the apoptosis and expression of proinflammatory cytokines in human osteoarthritis chondrocytes , 2017, Cytokine.

[3]  N. Lane,et al.  Osteoarthritis year in review 2016: clinical. , 2017, Osteoarthritis and cartilage.

[4]  Zhe Chen,et al.  AntimiR-30b Inhibits TNF-α Mediated Apoptosis and Attenuated Cartilage Degradation through Enhancing Autophagy , 2016, Cellular Physiology and Biochemistry.

[5]  Yadi Wu,et al.  Long Noncoding RNA UFC1 Promotes Proliferation of Chondrocyte in Osteoarthritis by Acting as a Sponge for miR-34a. , 2016, DNA and cell biology.

[6]  Jian-Ping Liu,et al.  The Importance of Toll‐like Receptors in NF‐κB Signaling Pathway Activation by Helicobacter pylori Infection and the Regulators of this Response , 2016, Helicobacter.

[7]  M. Valvano,et al.  Lipopolysaccharide modification in Gram-negative bacteria during chronic infection. , 2016, FEMS microbiology reviews.

[8]  Zhanguo Zhang,et al.  MicroRNA-630 suppresses tumor metastasis through the TGF-β- miR-630-Slug signaling pathway and correlates inversely with poor prognosis in hepatocellular carcinoma , 2016, Oncotarget.

[9]  J. Xu,et al.  A Human Anti-Toll Like Receptor 4 Fab Fragment Inhibits Lipopolysaccharide-Induced Pro-Inflammatory Cytokines Production in Macrophages , 2016, PloS one.

[10]  Cheng Yang,et al.  MicroRNA-146a Induced by Hypoxia Promotes Chondrocyte Autophagy through Bcl-2 , 2015, Cellular Physiology and Biochemistry.

[11]  N. Paschos.  Recent advances and future directions in the management of knee osteoarthritis: Can biological joint reconstruction replace joint arthroplasty and when? , 2015, World journal of orthopedics.

[12]  Y. Nie,et al.  CD14 knockdown reduces lipopolysaccharide-induced cell viability and expression of inflammation-associated genes in gastric cancer cells in vitro and in nude mouse xenografts. , 2015, Molecular medicine reports.

[13]  Xin-Rong Zhan,et al.  MiR-301a promotes cell proliferation by directly targeting TIMP2 in multiple myeloma. , 2015, International journal of clinical and experimental pathology.

[14]  Bing Zhang,et al.  The Involvement of Mutual Inhibition of ERK and mTOR in PLCγ1-Mediated MMP-13 Expression in Human Osteoarthritis Chondrocytes , 2015, International journal of molecular sciences.

[15]  T. Schnitzer,et al.  Efficacy and safety of tanezumab monotherapy or combined with non-steroidal anti-inflammatory drugs in the treatment of knee or hip osteoarthritis pain. , 2015, Annals of the rheumatic diseases.

[16]  Xu Wang,et al.  Clinical significance of high expression of circulating serum lncRNA RP11-445H22.4 in breast cancer patients: a Chinese population-based study , 2015, Tumor Biology.

[17]  P. Angrand,et al.  The role of long non-coding RNAs in genome formatting and expression , 2015, Front. Genet..

[18]  C. Labbaye,et al.  miR-146a controls CXCR4 expression in a pathway that involves PLZF and can be used to inhibit HIV-1 infection of CD4(+) T lymphocytes. , 2015, Virology.

[19]  Nan Xu,et al.  Microarray expression profile analysis of long non-coding RNAs in human breast cancer: a study of Chinese women. , 2015, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[20]  J. Xiang,et al.  MiR-301a Promotes Colorectal Cancer Cell Growth and Invasion by Directly Targeting SOCS6 , 2015, Cellular Physiology and Biochemistry.

[21]  Hee-Woo Lee,et al.  Anti-inflammatory effects of galangin on lipopolysaccharide-activated macrophages via ERK and NF-κB pathway regulation , 2014, Immunopharmacology and immunotoxicology.

[22]  Li-Yan Xu,et al.  Identification of Long Noncoding RNA Associated with Osteoarthritis in Humans , 2014, Orthopaedic surgery.

[23]  Xin Zhang,et al.  Long Noncoding RNA Related to Cartilage Injury Promotes Chondrocyte Extracellular Matrix Degradation in Osteoarthritis , 2014, Arthritis & rheumatology.

[24]  M. He,et al.  miR-9 targets CXCR4 and functions as a potential tumor suppressor in nasopharyngeal carcinoma. , 2014, Carcinogenesis.

[25]  Wentao Wang,et al.  miR-133b, a muscle-specific microRNA, is a novel prognostic marker that participates in the progression of human colorectal cancer via regulation of CXCR4 expression , 2013, Molecular Cancer.

[26]  Y. Okada,et al.  [Destruction of the articular cartilage in osteoarthritis]. , 2013, Clinical calcium.

[27]  E. Maiello,et al.  Chemokine receptor CXCR4: role in gastrointestinal cancer. , 2013, Critical reviews in oncology/hematology.

[28]  A. Papavassiliou,et al.  The NF-κB signalling pathway in osteoarthritis. , 2013, The international journal of biochemistry & cell biology.

[29]  Yin Liu,et al.  Down‐regulation of miR‐301a suppresses pro‐inflammatory cytokines in Toll‐like receptor‐triggered macrophages , 2013, Immunology.

[30]  Y. Qu,et al.  Decrease of miR-202-3p Expression, a Novel Tumor Suppressor, in Gastric Cancer , 2013, PloS one.

[31]  T. Roger,et al.  The sirtuin inhibitor cambinol impairs MAPK signaling, inhibits inflammatory and innate immune responses and protects from septic shock. , 2013, Biochimica et biophysica acta.

[32]  J. Ju,et al.  Overexpressed miR-301a promotes cell proliferation and invasion by targeting RUNX3 in gastric cancer , 2013, Journal of Gastroenterology.

[33]  W. Grizzle,et al.  CXCL12/CXCR4 Protein Signaling Axis Induces Sonic Hedgehog Expression in Pancreatic Cancer Cells via Extracellular Regulated Kinase- and Akt Kinase-mediated Activation of Nuclear Factor κB , 2012, The Journal of Biological Chemistry.

[34]  W. Richter,et al.  Regulation of H19 and its encoded microRNA-675 in osteoarthritis and under anabolic and catabolic in vitro conditions , 2012, Journal of Molecular Medicine.

[35]  W. Grizzle,et al.  CXCL12/CXCR4 signaling axis induces SHH expression in pancreatic cancer cells via ERK- and Akt- mediated activation of NF-κB: implications for bidirectional tumor-stromal interactions , 2012 .

[36]  H. Benghuzzi,et al.  Role of plant-derived antioxidants on NF-kb expression in LPS-stimulated macrophages - biomed 2011. , 2011, Biomedical sciences instrumentation.

[37]  Cheng-chao Sun,et al.  MicroRNA-155 Silencing Enhances Inflammatory Response and Lipid Uptake in Oxidized Low-Density Lipoprotein-Stimulated Human THP-1 Macrophages , 2010, Journal of Investigative Medicine.

[38]  P. Bullough,et al.  Atlas of Osteoarthritis , 1984 .