LncRNA KCNQ1OT1 acting as a ceRNA for miR-4458 enhances osteosarcoma progression by regulating CCND2 expression

  • Meng WangEmail author
  • Zengtao Wang
  • Xiaolei Zhu
  • Shibing Guan
  • Zhibo LiuEmail author


Osteosarcoma is prevalent worldwide and characterized as a challenging health burden. It has been increasingly indicated that long non-coding RNAs (lncRNAs) are significant in pathological processes of numerous cancers, exerting oncogenic or tumor-suppressive function. However, the participation of KCNQ1OT1 in osteosarcoma has not been elaborated. In this study, we focus on interrogating the function of KCNQ1OT1 and its underlying mechanism in osteosarcoma. Our work demonstrated the upregulation of KCNQ1OT1 in osteosarcoma through qRT-PCR. Besides, loss of function assay (CCK-8, transwell migration) indicated KCNQ1OT1 promoted cell proliferation, migration in osteosarcoma. Mechanically, KCNQ1OT1 acting as sponge for miR-4458 antagonized its tumor-suppressive impact on CCND2 expression. The anti-apoptotic nature of KCNQ1OT1 was also unveiled via caspase-3 activity assay. Overexpressed KCNQ1OT1 acted as competing endogenous RNA (ceRNA) for miR-4458 and subsequently reinforced target gene CCND2. Collectively, the results of rescue experiments suggested that the oncogenic role of KCNQ1OT1 was performed through sponging miR-4458 and upregulating CCND2 during osteosarcoma development, providing a novel perspective of intervention in osteosarcoma management.


KCNQ1OT1 miR-4458 CCND2 Osteosarcoma 



We are thankful to all who participated in this study.

Compliance with ethical standards

The Research Ethics Committee of this hospital gave consent to this study. Prior to diagnosis, none of patients were subjected to antitumor treatment. The written consents were eligibly gained from OS patients and in line with the Declaration of Helsinki, the study was implemented.

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

11626_2019_386_Fig5_ESM.png (216 kb)
Figure S1

(A) KCNQ1OT1 and CCND2 levels in OS tissues and non-OS tissues were examined via qRT-PCR analysis. (PNG 216 kb)

11626_2019_386_MOESM1_ESM.tif (6.4 mb)
High resolution image (TIF 6551 kb)


  1. Ast V, Kordaß T, Oswald M, Kolte A, Eisel D, Osen W, Eichmüller SB, Berndt A, König R (2018) MiR-192, miR-200c and miR-17 are fibroblast-mediated inhibitors of colorectal cancer invasion. Oncotarget 9:35559–35580CrossRefGoogle Scholar
  2. Cai Y, Wan J (2018) Competing endogenous RNA regulations in neurodegenerative disorders: current challenges and emerging insights. Front Mol Neurosci 11:370CrossRefGoogle Scholar
  3. Chen M, Fan L, Zhang SM, Li Y, Chen P, Peng X, Liu DB, Ma C, Zhang WJ, Zou ZW, Li PD (2019) LINC01939 inhibits the metastasis of gastric cancer by acting as a molecular sponge of miR-17-5p to regulate EGR2 expression. Cell Death Dis 10:70CrossRefGoogle Scholar
  4. Cheng C, Lu H, Wang Y (2016) miR-4458 suppresses glycolysis and lactate production by directly targeting hexokinase2 in colon cancer cells. Biochem Biophys Res Commun 469:37–43CrossRefGoogle Scholar
  5. Cheng D, Qiu X, Zhuang M, Liu Z (2018) Long noncoding RNA SNHG16 promotes cell proliferation by sponging MicroRNA-205 and upregulating ZEB1 expression in osteosarcoma. Cell Physiol Biochem 51:429–440CrossRefGoogle Scholar
  6. Flamini V, Dudley E, Jiang WG, Cui Y (2018) Distinct mechanisms by which two forms of miR-140 suppress the malignant properties of lung cancer cells. Oncotarget 9:36474–36491CrossRefGoogle Scholar
  7. Han W, Wang L, Zhang L, Wang Y, Li Y (2019) Circular RNA circ-RAD23B promotes cell growth and invasion by miR-593-3p/CCND2 and miR-653-5p/TIAM1 pathways in non-small cell lung cancer. Biochem Biophys Res Commun 510:462–466CrossRefGoogle Scholar
  8. Isakoff MS, Bielack SS, Meltzer P, Gorlick R (2015) Osteosarcoma: current treatment and a collaborative pathway to success. J Clin Oncol 33:3029–3035CrossRefGoogle Scholar
  9. Jia ZW, Li Y, Cui GR, Zhao HB, Li PY, Luo JM (2018) Expression and clinical significance of LncRNA KCNQ1OT1 in patients with acute myeloid leukemia. Zhongguo Shi Yan Xue Ye Xue Za Zhi 26:653–657PubMedGoogle Scholar
  10. Liang H, Yu T, Han Y, Jiang H, Wang C, You T, Zhao X, Shan H, Yang R, Yang L, Shan H, Gu Y (2018) LncRNA PTAR promotes EMT and invasion-metastasis in serous ovarian cancer by competitively binding miR-101-3p to regulate ZEB1 expression. Mol Cancer 17:119CrossRefGoogle Scholar
  11. Lv D-S, Li M, Sun G, Zhang X-F, Bai Y (2017) MicroRNA-4458 suppresses the proliferation of human lung cancer cells in vitro by directly targeting Lin28B. Acta Pharmacol Sin 38:1297–1304CrossRefGoogle Scholar
  12. Rui X, Xu Y, Jiang X, Ye W, Huang Y, Jiang J (2018) Long non-coding RNA C5orf66-AS1 promotes cell proliferation in cervical cancer by targeting miR-637/RING1 axis. Cell Death Dis 9:1175CrossRefGoogle Scholar
  13. Shen F, Chang H, Gao G, Zhang B, Li X, Jin B (2018) Long noncoding RNA FOXD2-AS1 promotes glioma malignancy and tumorigenesis via targeting miR-185-5p/CCND2 axis. J Cell Biochem 120:9324–9336. CrossRefPubMedGoogle Scholar
  14. Silva JM, Rodrigues S, Sampaio-Marques B, Gomes P, Neves-Carvalho A, Dioli C, Soares-Cunha C, Mazuik BF, Takashima A, Ludovico P, Wolozin B, Sousa N, Sotiropoulos I (2018) Dysregulation of autophagy and stress granule-related proteins in stress-driven Tau pathology. Cell Death Differ.
  15. Sio TT, Vu CC, Sohawon S, Van Houtte P, Thariat J, Novotny PJ, Miller RC, Bar-Sela G (2016) Extraskeletal osteosarcoma: an international rare cancer network study. Am J Clin Oncol 39:32–36CrossRefGoogle Scholar
  16. Soleimani M, Mandal SS (2017) Long noncoding RNA and cancer: a new paradigm. Cancer Res 77:3965–3981CrossRefGoogle Scholar
  17. Song N, Zhong J, Hu Q, Gu T, Yang B, Zhang J, Yu J, Ma X, Chen Q, Qi J, Liu Y, Su W, Feng Z, Wang X, Wang H (2018) FGF18 enhances migration and the epithelial-mesenchymal transition in breast cancer by regulating Akt/GSK3β/Β-catenin signaling. Cell Physiol Biochem 49:1019–1032CrossRefGoogle Scholar
  18. Su X, Teng J, Jin G, Li J, Zhao Z, Cao X, Guo Y, Guo M, Li X, Wu J, Wang C, Guo Z, Guo Q (2019) ELK1-induced upregulation of long non-coding RNA MIR100HG predicts poor prognosis and promotes the progression of osteosarcoma by epigenetically silencing LATS1 and LATS2. Biomed Pharmacother 109:788–797CrossRefGoogle Scholar
  19. Sun B, Yu H, Yang Z, Zhu L (2015) Tumor-suppressing effect of miR-4458 on human hepatocellular carcinoma. Cell Physiol Biochem 35:1797–1807CrossRefGoogle Scholar
  20. Tang D, Sun B, Yu H, Yang Z, Zhu L (2015) Tumor-suppressing effect of miR-4458 on human hepatocellular carcinoma. Cell Physiol Biochem 35:1797–1807CrossRefGoogle Scholar
  21. Wagner F, Holzapfel BM, McGovern JA, Shafiee A, Baldwin JG, Martine LC, Lahr CA, Wunner FM, Friis T, Bas O, Boxberg M, Prodinger PM, Shokoohmand A, Moi D, Mazzieri R, Loessner D, Hutmacher DW (2018) Humanization of bone and bone marrow in an orthotopic site reveals new potential therapeutic targets in osteosarcoma. Biomaterials 171:230–246CrossRefGoogle Scholar
  22. Wang C, Liang C, Yang H, Chen D, Yu X, Zhao W, Geng D, Li S, Chen Z, Sun M (2018a) The dysregulated expression of KCNQ1OT1 and its interaction with downstream factors miR-145/CCNE2 in breast cancer cells. Cell Physiol Biochem 49:432–446CrossRefGoogle Scholar
  23. Wang D, Niu X, Wang Z, Song CL, Huang Z, Chen KN, Duan J, Bai H, Xu J, Zhao J, Wang Y, Zhuo M, Xie XS, Kang X, Tian Y, Cai L, Han JF, An T, Sun Y, Gao S, Zhao J, Ying J, Wang L, He J, Wang J (2019) Multiregion sequencing reveals the genetic heterogeneity and evolutionary history of osteosarcoma and matched pulmonary metastases. Cancer Res 79:7–20CrossRefGoogle Scholar
  24. Wang LY, Yu YL, Chen HW, Srivastava S, Petrovics G, Kung HJ (2014) A long noncoding RNA connects c-Myc to tumor metabolism. Proc Natl Acad Sci U S A 111:18697–18702CrossRefGoogle Scholar
  25. Wang Z, Wu H, Xue M, Lin P, Wang S, Lin N, Huang X, Pan W, Liu M, Yan X, Qu H, Sun L, Li H, Wu Y, Teng W, Wang Z, Zhou X, Chen H, Poznansky MC, Ye Z (2018b) Epigenetic regulation of CXCL12 plays a critical role in mediating tumor progression and the immune response in osteosarcoma. Cancer Res 78:3938–3953CrossRefGoogle Scholar
  26. Wong KHY, Levy-Sakin M, Kwok P-Y (2018) De novo human genome assemblies reveal spectrum of alternative haplotypes in diverse populations. Nat Commun 9:3040CrossRefGoogle Scholar
  27. Xie Y, Sundström A, Maturi NP, Tan EJ, Marinescu VD, Jarvius M, Tirfing M, Jin C, Chen L, Essand M, Swartling FJ, Nelander S, Jiang Y, Uhrbom L (2018) LGR5 promotes tumorigenicity and invasion of glioblastoma stem-like cells and is a potential therapeutic target for a subset of glioblastoma patients. J Pathol 247:228–240. CrossRefGoogle Scholar
  28. Xu M, Chen X, Lin K, Zeng K, Liu X, Xu X, Pan B, Xu T, Sun L, He B, Pan Y, Sun H, Wang S (2019) lncRNA SNHG6 regulates EZH2 expression by sponging miR-26a/b and miR-214 in colorectal cancer. J Hematol Oncol 12:3CrossRefGoogle Scholar
  29. Xu R, Huang J, Zhao J, Shi W (2017) Knockdown of long non-coding RNA KCNQ1OT1 depressed chemoresistance to paclitaxel in lung adenocarcinoma. Cancer Chemother Pharmacol 80:243–250CrossRefGoogle Scholar
  30. Yao Y, Zhou Y, Fu X (2019) miR6713p is downregulated in nonsmall cell lung cancer and inhibits cancer progression by directly targeting CCND2. Mol Med Rep 19:2407–2412PubMedGoogle Scholar
  31. Yu HM, Wang C, Yuan Z, Chen GL, Ye T, Yang BW (2018) LncRNA NEAT1 promotes the tumorigenesis of colorectal cancer by sponging miR-193a-3p. Cell Prolif.
  32. Yu W, Wang Y, Zhu J, Jin L, Liu B, Xia K, Wang J, Gao J, Liang C, Tao H (2019) Autophagy inhibitor enhance ZnPc/BSA nanoparticle induced photodynamic therapy by suppressing PD-L1 expression in osteosarcoma immunotherapy. Biomaterials 192:128–139CrossRefGoogle Scholar
  33. Zhang Y, Yang G, Luo Y (2019) Long non-coding RNA PVT1 promotes glioma cell proliferation and invasion by targeting miR-200a. Exp Ther Med 17:1337–1345PubMedGoogle Scholar
  34. Zhao W, Li L (2019) SP1-induced upregulation of long non-coding RNA HCP5 promotes the development of osteosarcoma. Pathol Res Pract 215:439–445CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  1. 1.Hand and Foot Surgical CenterProvincial Hospital Affiliated to Shandong UniversityJinanChina

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