LncRNA expression profile analysis of Mg2+-induced osteogenesis by RNA-seq and bioinformatics

다국어 초록 (Multilingual Abstract)

Background In recent years, magnesium (Mg) has been extensively studied for manufacturing biodegradable orthopedic devices. Besides other advantages, researches have shown that magnesium-based implants can stimulate osteogenesis thus accelerating orthopedic trauma recovery, but its molecular mechanism is not fully understood. Meanwhile, long non-coding RNA (lncRNA) has been found to play vital role in regulating osteogenic diferentiation.
Objective To explore the role of lncRNA in Mg2+ (magnesium ions)-induced osteogenesis.
Methods The efect of Mg2+ on mBMSCs proliferation was detected by the CCK-8 assay. The optimum concentration of Mg2+ (7.5 mM) in promoting mBMSCs osteogenesis was determined by ALP staining and Alizarin red staining, western blot and RT-qPCR were performed to detect osteogenic markers expressions. The lncRNAs and mRNAs expression profles of mBMSCs were assessed by RNA-Seq and processed by bioinformatics analysis. The selected lncRNAs expression level was validated by RT-qPCR.
Results The efect of Mg2+ in promoting osteogenesis was confrmed and the optimum concentration was determined as 7.5 mM. The lncRNAs and mRNAs diferentially expressed between 7.5 mM Mg2+-treated group and control group was detected and functional analysis revealed that their function were associated with osteogenesis. The ceRNA networks were constructed for H19 and Dubr that aberrantly expressed in two groups. The ceRNA networks of selected lncRNAs (H19 and Dubr) were constructed.
Conclusions This study identifed H19 and Dubr as osteogenic associated lncRNAs involved in Mg2+-induced osteogenesis, and they might play their roles through lncRNA-miRNA–mRNA axis.

참고문헌 (Reference)

1 Ulitsky I, "lincRNAs : genomics, evolution, and mechanisms" 154 : 26-46, 2013

2 Pertea M, "Transcript-level expression analysis of RNA-seq experiments with HISAT, StringTie and Ballgown" 11 : 1650-1667, 2016

3 Ko NY, "The role of micro RNA and longnon-coding RNA in osteoporosis" 21 : 4886-4903, 2020

4 Hung CC, "The role of magnesium ions in bone regeneration involves the canonical Wnt signaling pathway" 98 : 246-255, 2019

5 Liu Y, "The role of long non-coding RNA H19in musculoskeletal system : a new player in an old game" 360 : 61-65, 2017

6 Nakashima K, "The novel zinc fnger-containing transcription factor osterix is required for osteoblast diferentiation and bone formation" 108 : 17-29, 2002

7 Witte F, "The history of biodegradable magnesium implants : a review" 6 : 1680-1692, 2010

8 Song W, "The emerging roles of long noncoding RNAs in bone homeostasis and their potential application in bone-related diseases" 39 : 926-937, 2020

9 Bruderer M, "Role and regulation of RUNX2 in osteogenesis" 28 : 269-286, 2014

10 Zhang Y, "Research progress regarding the role of long non-coding RNAs in osteosarcoma" 20 : 2606-2612, 2020

11 Gomathi K, "Regulation of Runx2 by post-translational modifcations in osteoblast diferentiation" 245 : 117389-, 2020

12 Steck E, "Regulation of H19 and its encoded microRNA-675 in osteoarthritis and under anabolic and catabolic in vitro conditions" 90 : 1185-1195, 2012

13 Tian Y, "Osterix is required for Sonic hedgehog-induced osteoblastic MC3T3-E1 cell diferentiation" 64 : 169-176, 2012

14 Iwayama T, "Osteoblastic lysosome plays a central role in mineralization" 5 : eaax0672-, 2019

15 Jimi E, "NF-κB signaling regulates physiological and pathological chondrogenesis" 20 : 6275-6285, 2019

16 Guttman M, "Modular regulatory principles of large non-coding RNAs" 482 : 339-346, 2012

17 Wang Z, "Mg(2+)in beta-TCP/Mg-Zn composite enhances the diferentiation of human bone marrow stromal cells into osteoblasts through MAPK-regulated Runx2/Osx" 235 : 5182-5191, 2020

18 Cuadrado A, "Mechanisms and functions of p38MAPK signalling" 429 : 403-417, 2010

19 Qiao X, "Magnesium-doped nanostructured titanium surface modulates macrophage-mediated infammatory response for ameliorative osseointegration" 15 : 7185-7198, 2020

20 Yoshizawa S, "Magnesium ion stimulation of bone marrow stromal cells enhances osteogenic activity, simulating the efect of magnesium alloy degradation" 10 : 2834-2842, 2014

21 Hu T, "Magnesium enhances the chondrogenic diferentiation of mesenchymal stem cells by inhibiting activated macrophage-induced infammation" 8 : 3406-, 2018

22 Diaz-Tocados JM, "Magnesium chloride promotes osteogenesis through notch signaling activation and expansion of mesenchymal stem cells" 7 : 7839-, 2017

23 Staiger MP, "Magnesium and its alloys as orthopedic biomaterials : a review" 27 : 1728-1734, 2006

24 Yang Q, "Long noncoding RNAs : new players in the osteogenic diferentiation of bone marrow-and adipose-derived mesenchymal stem cells" 14 : 297-308, 2018

25 Yu X, "Long noncoding RNA Taurine upregulated gene 1 promotes osteosarcoma cell metastasis by mediating HIF-1α via miR-143-5p" 10 : 280-, 2019

26 Wang Y, "Long noncoding RNA H19 mediates LCoR to impact the osteogenic and adipogenic diferentiation of mBMSCs in mice through sponging miR-188" 233 : 7435-7446, 2018

27 Huang XZ, "LncRNA-MALAT1 promotes osteogenic diferentiation through regulating ATF4 by sponging miR-214 : implication of steroid-induced avascular necrosis of the femoral head" 154 : 108533-, 2020

28 Zhang N, "LncRNA MSC-AS1 promotes osteogenic differentiation and alleviates osteoporosis through sponging microRNA-140-5p to upregulate BMP2" 519 : 790-796, 2019

29 Gao X, "LncRNA KCNQ1OT1 promotes osteogenic diferentiation to relieve osteolysis via Wnt/βcatenin activation" 8 : 19-, 2018

30 Wang L, "LncRNA Dum interacts with Dnmts to regulate Dppa2 expression during myogenic differentiation and muscle regeneration" 25 : 335-350, 2015

31 Chen LL, "Linking long noncoding RNA localization and function" 41 : 761-772, 2016

32 Feng L, "Linc-ROR promotes osteogenic differentiation of mesenchymal stem cells by functioning as a competing endogenous RNA for miR-138 and miR-145" 11 : 345-353, 2018

33 Djebali S, "Landscape of transcription in human cells" 489 : 101-108, 2012

34 Levasseur R, "LRP5 mutations in osteoporosis-pseudoglioma syndrome and high-bone-mass disorders" 72 : 207-214, 2005

35 Williams BO, "LRP5 : From bedside to bench to bone" 102 : 26-30, 2017

36 Cabili MN, "Integrative annotation of human large intergenic noncoding RNAs reveals global properties and specific subclasses" 25 : 1915-1927, 2011

37 Huo S, "Insight into the role of long non-coding RNAs during osteogenesis in mesenchymal stem cells" 13 : 52-59, 2018

38 Witte F, "In vivo corrosion and corrosion protection of magnesium alloy LAE442" 6 : 1792-1799, 2010

39 Zhang Y, "Implant-derived magnesium induces local neuronal production of CGRP to improve bonefracture healing in rats" 22 : 1160-1169, 2016

40 Hong S, "Identifcation of functional lncRNAs based on competing endogenous RNA network in osteoblast diferentiation" 235 : 2232-2244, 2020

41 Maradze D, "High magnesium corrosion rate has an efect on osteoclast and mesenchymal stem cell role during bone remodelling" 8 : 10003-, 2018

42 Liang WC, "H19 activates Wnt signaling and promotes osteoblast diferentiation by functioning as a competing endogenous RNA" 6 : 20121-, 2016

43 Ponting CP, "Evolution and functions of long noncoding RNAs" 136 : 629-641, 2009

44 Liu C, "Enhanced osteoinductivity and corrosion resistance of dopamine/gelatin/rhBMP-2-coated beta-TCP/Mg-Zn orthopedic implants : an in vitro and in vivo study" 15 : e0228247-, 2020

45 Wang L, "Diferential expression of long noncoding ribonucleic acids during osteogenic diferentiation of human bone marrow mesenchymal stem cells" 39 : 1013-1019, 2015

46 Zhao D, "Current status on clinical applications of magnesium-based orthopaedic implants : a review from clinical translational perspective" 112 : 287-302, 2017

47 Ala U, "Competing endogenous RNAs, non-coding RNAs and diseases : an intertwined story" 9 (9): 1574-, 2020

48 Han HS, "Biodegradable magnesium alloys promote angio-osteogenesis to enhance bone repair" 7 : 2000800-, 2020

49 Agarwal S, "Biodegradable magnesium alloys for orthopaedic applications : a review on corrosion, biocompatibility and surface modifcations" 68 : 948-963, 2016

50 Kang Z, "Bio-based strategies for producing glycosaminoglycans and their oligosaccharides" 36 : 806-818, 2018

51 Alipoor B, "An updated review of the H19 lncRNA in human cancer : molecular mechanism and diagnostic and therapeutic importance" 47 : 6357-6374, 2020

52 Peng S, "An overview of long noncoding RNAs involved in bone regeneration from mesenchymal stem cells" 2018 : 8273648-, 2018

53 Consortium EP, "An integrated encyclopedia of DNA elements in the human genome" 489 : 57-74, 2012

54 Salmena L, "A ceRNA hypothesis: the Rosetta stone of a hidden RNA language?" 146 : 353-358, 2011

55 Mansour A, "(*)Extracellular matrices for bone regeneration : a literature review" 23 : 1436-1451, 2017