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Endocrine

, 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 Zhang
  • Ping Zhang
  • Zhihong Wang
  • Wenwu Dong
  • Liang He
  • Ting Zhang
  • Liang Shao
  • Wenqian Zhang
  • Changhao Wu
Original Article

Abstract

Purpose

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.

Methods

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.

Results

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.

Conclusions

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.

Keywords

lncRNA GAS8-AS1 thyroid cancer autophagy 

Notes

Funding

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.

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Copyright information

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

Authors and Affiliations

  • Yuan Qin
    • 1
  • Wei Sun
    • 1
  • Hao Zhang
    • 1
  • 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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