, Volume 59, Issue 3, pp 555–564 | Cite as

LncRNA GAS8-AS1 inhibits cell proliferation through ATG5-mediated autophagy in papillary thyroid cancer

  • Yuan Qin
  • Wei Sun
  • Hao ZhangEmail author
  • Ping Zhang
  • Zhihong Wang
  • Wenwu Dong
  • Liang He
  • Ting Zhang
  • Liang Shao
  • Wenqian Zhang
  • Changhao Wu
Original Article



The long non-coding RNA GAS8 antisense RNA 1 (lncRNA GAS8-AS1) is a tumor suppressor in papillary thyroid cancer (PTC), but the mechanisms underlying how GAS8-AS1 regulates PTC biology remain unclear. Here, we evaluated the molecular function of GAS8-AS1 in regulating autophagy in PTC cell lines.


GAS8-AS1 was overexpressed and knocked down in PTC cell lines by transfecting with expression plasmids or short interfering RNAs (siRNAs). Cell proliferation was evaluated using the Cell Counting Kit-8 (CCK-8). qRT-PCR and western blot were used to determine changes in expression of autophagy-related genes. Autophagy was evaluated by immunofluorescence and transmission electron microscopy.


Relative GAS8-AS1 expression was lower in the PTC cell lines, TPC1 and BCPAP, compared to a normal thyroid cell line. Overexpression of GAS8-AS1 inhibited proliferation, significantly increased the ratio of LC3-II/LC3-I, and reduced p62 expression, whereas GAS8-AS1 knockdown demonstrated opposite effects. In GAS8-AS1 overexpressing cell lines, LC3 immunofluorescence staining demonstrated increased punctate aggregates of LC3 staining, and transmission electron microscopy revealed increased numbers of autophagosomes. Autophagy-related gene 5 (ATG5) was markedly upregulated by GAS8-AS1 overexpression and downregulated by GAS8-AS1 knockdown. Finally, silencing of ATG5 attenuated autophagy activation and rescued the inhibition of cell proliferation caused by GAS8-AS1.


In PTC cell lines, GAS8-AS1 inhibited proliferation, activated autophagy, and increased ATG5 expression. Downregulation of ATG5 reversed GAS8-AS1-mediated activation of autophagy leading to cell death, revealing a novel mechanism of the GAS8-AS1-ATG5 axis in PTC cell lines. This provided a new experimental basis to explore the effects of lncRNA on autophagy in the treatment of thyroid cancer.


lncRNA GAS8-AS1 thyroid cancer autophagy 



This study was funded by the Natural Science Foundation of Liaoning Province (No. 2015020536), Liaoning BaiQianWan Talents Program (No. 2014921033), Science and Technology Project of Shenyang City (No. F16-205-1-41).

Compliance with ethical standards

Conflict of interest

The authors have full control of all of the primary data and agree to allow the journal to review the data if needed. The authors declare that they have no competing interests.

Ethical approval

This article does not contain any studies with human participants or animals performed by any of the authors.


  1. 1.
    G. Pellegriti, F. Frasca, C. Regalbuto, S. Squatrito, R. Vigneri, Worldwide increasing incidence of thyroid cancer: update on epidemiology and risk factors. J. Cancer Epidemiol. 2013, 965212 (2013)CrossRefGoogle Scholar
  2. 2.
    O. Gimm, M.D. Castellone, C. Hoang-Vu, E. Kebebew, Biomarkers in thyroid tumor research: new diagnostic tools and potential targets of molecular-based therapy. J. Thyroid. Res. 2011, 631593 (2011)CrossRefGoogle Scholar
  3. 3.
    R.W. Randle et al., Trends in the presentation, treatment, and survival of patients with medullary thyroid cancer over the past 30 years. Surgery 161, 137–146 (2017)CrossRefGoogle Scholar
  4. 4.
    D.F. Schneider, H. Chen, New developments in the diagnosis and treatment of thyroid cancer. CA Cancer J. Clin. 63, 374–394 (2013)CrossRefGoogle Scholar
  5. 5.
    L. Lamartina, et al., Risk stratification of neck lesions detected sonographically during the follow-up of differentiated thyroid cancer. J. Clin. Endocrinol. Metab 101, 3036–3044 (2016)CrossRefGoogle Scholar
  6. 6.
    C.M. Kitahara, J.A. Sosa, The changing incidence of thyroid cancer. Nat. Rev. Endocrinol. 12, 646–653 (2016)CrossRefGoogle Scholar
  7. 7.
    C.K. Chou, R.T. Liu, H.Y. Kang, MicroRNA-146b: a novel biomarker and therapeutic target for human papillary thyroid cancer. Int. J. Mol. Sci. 18, E636 (2017)CrossRefGoogle Scholar
  8. 8.
    B.R. Haugen et al., 2015 American Thyroid Association Management Guidelines for adult patients with thyroid nodules and differentiated thyroid cancer: The American Thyroid Association Guidelines Task Force on Thyroid Nodules and Differentiated Thyroid Cancer. Thyroid 26, 1–133 (2016)CrossRefGoogle Scholar
  9. 9.
    J.T. Lee, Epigenetic regulation by long noncoding RNAs. Science 338, 1435–1439 (2012)CrossRefGoogle Scholar
  10. 10.
    J.L. Rinn, H.Y. Chang, Genome regulation by long noncoding RNAs. Annu. Rev. Biochem. 81, 145–166 (2012)CrossRefGoogle Scholar
  11. 11.
    J. Zhao, Y. Liu, G. Huang, P. Cui, W. Zhang, Y. Zhang, Long non-coding RNAs in gastric cancer: versatile mechanisms and potential for clinical translation. Am. J. Cancer Res.. 5, 907–927 (2015)PubMedPubMedCentralGoogle Scholar
  12. 12.
    J. Sun et al., Inferring novel lncRNA-disease associations based on a random walk model of a lncRNA functional similarity network. Mol. Biosyst. 10, 2074–2081 (2014)CrossRefGoogle Scholar
  13. 13.
    R. Fatima, V.S. Akhade, D. Pal, S.M. Rao, Long noncoding RNAs in development and cancer: potential biomarkers and therapeutic targets. Mol. Cell. Ther. 3, 5 (2015)CrossRefGoogle Scholar
  14. 14.
    M. Zhou et al., Prioritizing candidate disease-related long non-coding RNAs by walking on the heterogeneous lncRNA and disease network. Mol. Biosyst. 11, 760–769 (2015)CrossRefGoogle Scholar
  15. 15.
    M. Zhou et al., A potential signature of eight long non-coding RNAs predicts survival in patients with non-small cell lung cancer. J. Transl. Med. 13, 231 (2015)CrossRefGoogle Scholar
  16. 16.
    M. Zhou et al., Comprehensive analysis of lncRNA expression profiles reveals a novel lncRNA signature to discriminate nonequivalent outcomes in patients with ovarian cancer. Oncotarget 7, 32433–32448 (2016)PubMedPubMedCentralGoogle Scholar
  17. 17.
    M. Zhou et al., Identification and validation of potential prognostic lncRNA biomarkers for predicting survival in patients with multiple myeloma. J. Exp. Clin. Cancer Res. 34, 102 (2015)CrossRefGoogle Scholar
  18. 18.
    H. Cai et al., LncRNA HOTAIR acts a competing endogenous RNA to control the expression of notch3 via sponging miR-613 in pancreatic cancer. Oncotarget 8, 32905–32917 (2017)PubMedPubMedCentralGoogle Scholar
  19. 19.
    L. Fang, et al., Long non-coding RNA NEAT1 promotes hepatocellular carcinoma cell proliferation through the regulation of miR-129-5p-VCP-IκB. Am. J. Physiol. Gastrointest. Liver Physiol 313, G150–G156 (2017)CrossRefGoogle Scholar
  20. 20.
    W. Zhou et al., The lncRNA H19 mediates breast cancer cell plasticity during EMT and MET plasticity by differentially sponging miR-200b/c and let-7b. Sci. Signal 10, eaak9557 (2017)CrossRefGoogle Scholar
  21. 21.
    P.D. Li et al., Upregulation of the long non-coding RNA PVT1 promotes esophageal squamous cell carcinoma progression by acting as a molecular sponge of miR-203 and LASP1. Oncotarget 8, 34164–34176 (2017)PubMedPubMedCentralGoogle Scholar
  22. 22.
    M. Yang et al., Long noncoding RNA are aberrantly expressed in human papillary thyroid carcinoma. Oncol. Lett. 12, 544–552 (2016)CrossRefGoogle Scholar
  23. 23.
    W. Pan et al., Whole exome sequencing identifies lncRNA GAS8-AS1 and LPAR4 as novel papillary thyroid carcinoma driver alternations. Hum. Mol. Genet. 25, 1875–1884 (2016)CrossRefGoogle Scholar
  24. 24.
    A.M. Choi, S.W. Ryter, B. Levine, Autophagy in human health and disease. N. Engl. J. Med. 368, 651–662 (2013)CrossRefGoogle Scholar
  25. 25.
    K. Wang et al., APF lncRNA regulates autophagy and myocardial infarction by targeting miR-188-3p. Nat. Commun. 6, 6779 (2015)CrossRefGoogle Scholar
  26. 26.
    Q.W. Fan et al., Akt and autophagy cooperate to promote survival of drug-resistant glioma. Sci. Signal 3, ra81 (2010)CrossRefGoogle Scholar
  27. 27.
    J.S. Carew et al., Cleveland, The novel polyamine analogue CGC-11093 enhances the antimyeloma activity of bortezomib. Cancer Res. 68, 4783–4790 (2008)CrossRefGoogle Scholar
  28. 28.
    Z.J. Yang, C.E. Chee, S. Huang, F.A. Sinicrope, The role of autophagy in cancer: therapeutic implications. Mol. Cancer Ther. 10, 1533–1541 (2011)CrossRefGoogle Scholar
  29. 29.
    B.P. Rubin, J. Debnath, Therapeutic implications of autophagy-mediated cell survival in gastrointestinal stromal tumor after treatment with imatinib mesylate. Autophagy 6, 1190–1191 (2010)CrossRefGoogle Scholar
  30. 30.
    D.J. Klionsky et al., Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition). Autophagy 12, 1–222 (2016)CrossRefGoogle Scholar
  31. 31.
    J. Lin et al., Inhibition of autophagy enhances the anticancer activity of silver nanoparticles. Autophagy 10, 2006–2020 (2014)CrossRefGoogle Scholar
  32. 32.
    T.R. Mercer, M.E. Dinger, J.S. Mattick, Long non-coding RNAs: insights into functions. Nat. Rev. Genet. 10, 155–159 (2009)CrossRefGoogle Scholar
  33. 33.
    J. Liz, M. Esteller, lncRNAs and microRNAs with a role in cancer development. Biochim. Biophys. Acta 1859, 169–176 (2016)CrossRefGoogle Scholar
  34. 34.
    H. Li, B. Yu, J. Li, L. Su, M. Yan, Z. Zhu, B. Liu, Overexpression of lncRNA H19 enhances carcinogenesis and metastasis of gastric cancer. Oncotarget 5, 2318–2329 (2014)PubMedPubMedCentralGoogle Scholar
  35. 35.
    T. Nakagawa et al., Large noncoding RNA HOTAIR enhances aggressive biological behavior and is associated with short disease-free survival in human non-small cell lung cancer. Biochem. Biophys. Res. Commun. 436, 319–324 (2013)CrossRefGoogle Scholar
  36. 36.
    W. Sun et al., Overexpression of long non-coding RNA NR_036575.1 contributes to the proliferation and migration of papillary thyroid cancer. Med. Oncol. 33, 102 (2016)CrossRefGoogle Scholar
  37. 37.
    X. Lan et al., Downregulation of long noncoding RNA NONHSAT037832 in papillary thyroid carcinoma and its clinical significance. Tumour Biol. 37, 6117–6123 (2016)CrossRefGoogle Scholar
  38. 38.
    D. Zhang, X. Liu, B. Wei, G. Qiao, T. Jiang, Z. Chen, Plasma lncRNA GAS8-AS1 as a potential biomarker of papillary thyroid carcinoma in Chinese patients. Int. J. Endocrinol. 2017, 2645904 (2017)PubMedPubMedCentralGoogle Scholar
  39. 39.
    F. Scarlatti, R. Granata, A.J. Meijer, P. Codogno, Does autophagy have a license to kill mammalian cells. Cell. Death Differ. 16, 12–20 (2009)CrossRefGoogle Scholar
  40. 40.
    M. Gugnoni et al., Cadherin-6 promotes EMT and cancer metastasis by restraining autophagy. Oncogene 36, 667–677 (2017)CrossRefGoogle Scholar
  41. 41.
    M. Hamada, H. Kameyama, S. Iwai, Y. Yura, Induction of autophagy by sphingosine kinase 1 inhibitor PF-543 in head and neck squamous cell carcinoma cells. Cell. Death Discov. 3, 17047 (2017)CrossRefGoogle Scholar
  42. 42.
    D.J. Klionsky et al., Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes. Autophagy 4, 151–175 (2008)CrossRefGoogle Scholar
  43. 43.
    N. Mizushima, T. Yoshimori, B. Levine, Methods in mammalian autophagy research. Cell 140, 313–326 (2010)CrossRefGoogle Scholar
  44. 44.
    Y.L. Xiu et al., Upregulation of the lncRNA Meg3 induces autophagy to inhibit tumorigenesis and progression of epithelial ovarian carcinoma by regulating activity of ATG3. Oncotarget 8, 31714–31725 (2017)PubMedPubMedCentralGoogle Scholar
  45. 45.
    L. Yu et al., Regulation of an ATG7-beclin 1 program of autophagic cell death by caspase-8. Science 304, 1500–1502 (2004)CrossRefGoogle Scholar
  46. 46.
    S. Shimizu et al., Role of Bcl-2 family proteins in a non-apoptotic programmed cell death dependent on autophagy genes. Nat. Cell. Biol. 6, 1221–1228 (2004)CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Yuan Qin
    • 1
  • Wei Sun
    • 1
  • Hao Zhang
    • 1
    Email author
  • Ping Zhang
    • 1
  • Zhihong Wang
    • 1
  • Wenwu Dong
    • 1
  • Liang He
    • 1
  • Ting Zhang
    • 1
  • Liang Shao
    • 1
  • Wenqian Zhang
    • 1
  • Changhao Wu
    • 1
  1. 1.Department of Thyroid SurgeryThe First Hospital of China Medical UniversityShenyangChina

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