Downregulation of LncRNA GAS5 promotes liver cancer proliferation and drug resistance by decreasing PTEN expression

  • Congren Wang
  • Shaoying Ke
  • Mingzhu Li
  • Conglin Lin
  • Xiaoyu Liu
  • Qunxiong PanEmail author
Original Article


Accumulating evidence has shown that the long noncoding RNAs (lncRNAs) play a crucial role in the regulation of hepatocellular carcinoma (HCC) progression and drug resistance. In this study, we aimed to investigate the biological function roles of lncRNAs growth arrest-specific 5 (GAS5) and its underlying molecular mechanism in the development of HCC. qRT-PCR was used to detect GAS5, miR-21, and PTEN levels. MTT, cell counting assays, and xenograft mouse model were applied to measure cell proliferation rate in vitro and in vivo. The luciferase reporter assay and RNA immune-precipitation assay were introduced to evaluate the relationship between GAS5 and miR-21. We found that GAS5 was downregulated in HCC cell lines and tumor tissues. Knockdown of GAS5 enhanced HCC cell proliferation in vitro and in vivo and increased HCC cell resistance to doxorubicin. GAS5 acted as a sponge for miR-21 silencing and consequently led to the elevation of PTEN expression. Our data demonstrated that GAS5 functioned as a tumor suppressor role in HCC through regulation of miR-21-PTEN singling pathways, suggesting a potential application of GAS5 in HCC therapy.


Hepatocellular carcinoma GAS5 miR-21 PTEN Tumor suppressor 




Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care were followed.

Informed consent

Informed consent was derived from all the participants.


  1. Avgeris M, Tsilimantou A, Levis PK, Tokas T, Sideris DC, Stravodimos K, Ardavanis A, Scorilas A (2018) Loss of GAS5 tumour suppressor lncRNA: an independent molecular cancer biomarker for short-term relapse and progression in bladder cancer patients. Br J Cancer 119(12):1477–1486CrossRefGoogle Scholar
  2. Balogh J, Victor D 3rd, Asham EH, Burroughs SG, Boktour M, Saharia A, Li X, Ghobrial RM, Monsour HP Jr (2016) Hepatocellular carcinoma: a review. J Hepatocell Carcinoma 3:41–53CrossRefGoogle Scholar
  3. Bruix J, Llovet JM (2002) Prognostic prediction and treatment strategy in hepatocellular carcinoma. Hepatology 35(3):519–524CrossRefGoogle Scholar
  4. Chen F, Li Y, Li M, Wang L (2019) Long noncoding RNA GAS5 inhibits metastasis by targeting miR-182/ANGPTL1 in hepatocellular carcinoma. Am J Cancer Res 9(1):108–121PubMedPubMedCentralGoogle Scholar
  5. Coccia EM, Cicala C, Charlesworth A, Ciccarelli C, Rossi GB, Philipson L, Sorrentino V (1992) Regulation and expression of a growth arrest-specific gene (gas5) during growth, differentiation, and development. Mol Cell Biol 12(8):3514–3521CrossRefGoogle Scholar
  6. Dhanasekaran R, Limaye A, Cabrera R (2012) Hepatocellular carcinoma: current trends in worldwide epidemiology, risk factors, diagnosis, and therapeutics. Hepat Med 4:19–37PubMedPubMedCentralGoogle Scholar
  7. DiStefano JK (2018) The emerging role of long noncoding RNAs in human disease. Methods Mol Biol 1706:91–110CrossRefGoogle Scholar
  8. Eissmann M, Gutschner T, Hammerle M, Gunther S, Caudron-Herger M, Gross M, Schirmacher P, Rippe K, Braun T, Zornig M, Diederichs S (2012) Loss of the abundant nuclear non-coding RNA MALAT1 is compatible with life and development. RNA Biol 9(8):1076–1087CrossRefGoogle Scholar
  9. El-Serag HB, Zhu AX, Johnson MS (2017) The treatment path in hepatocellular carcinoma. Clin Adv Hematol Oncol 15:1–20PubMedGoogle Scholar
  10. Fatica A, Bozzoni I (2014) Long non-coding RNAs: new players in cell differentiation and development. Nat Rev Genet 15(1):7–21CrossRefGoogle Scholar
  11. Gao Q, Xie H, Zhan H, Li J, Liu Y, Huang W (2017) Prognostic values of long noncoding RNA GAS5 in various carcinomas: an updated systematic review and meta-analysis. Front Physiol 8:814CrossRefGoogle Scholar
  12. Geisler S, Coller J (2013) RNA in unexpected places: long non-coding RNA functions in diverse cellular contexts. Nat Rev Mol Cell Biol 14(11):699–712CrossRefGoogle Scholar
  13. Ghouri YA, Mian I, Rowe JH (2017) Review of hepatocellular carcinoma: epidemiology, etiology, and carcinogenesis. J Carcinog 16:1CrossRefGoogle Scholar
  14. Gibb EA, Brown CJ, Lam WL (2011) The functional role of long non-coding RNA in human carcinomas. Mol Cancer 10:38CrossRefGoogle Scholar
  15. Hou Z, Xu X, Fu X, Tao S, Zhou J, Liu S, Tan D (2017) HBx-related long non-coding RNA MALAT1 promotes cell metastasis via up-regulating LTBP3 in hepatocellular carcinoma. Am J Cancer Res 7(4):845–856PubMedPubMedCentralGoogle Scholar
  16. Hu L, Ye H, Huang G, Luo F, Liu Y, Liu Y, Yang X, Shen J, Liu Q, Zhang J (2016) Long noncoding RNA GAS5 suppresses the migration and invasion of hepatocellular carcinoma cells via miR-21. Tumour Biol 37(2):2691–2702CrossRefGoogle Scholar
  17. Kallen AN, Zhou XB, Xu J, Qiao C, Ma J, Yan L, Lu L, Liu C, Yi JS, Zhang H, Min W, Bennett AM, Gregory RI, Ding Y, Huang Y (2013) The imprinted H19 lncRNA antagonizes let-7 microRNAs. Mol Cell 52(1):101–112CrossRefGoogle Scholar
  18. Kornienko AE, Guenzl PM, Barlow DP, Pauler FM (2013) Gene regulation by the act of long non-coding RNA transcription. BMC Biol 11:59CrossRefGoogle Scholar
  19. Lai MC, Yang Z, Zhou L, Zhu QQ, Xie HY, Zhang F, Wu LM, Chen LM, Zheng SS (2012) Long non-coding RNA MALAT-1 overexpression predicts tumor recurrence of hepatocellular carcinoma after liver transplantation. Med Oncol 29(3):1810–1816CrossRefGoogle Scholar
  20. Li C, Miao R, Liu S, Wan Y, Zhang S, Deng Y, Bi J, Qu K, Zhang J, Liu C (2017) Down-regulation of miR-146b-5p by long noncoding RNA MALAT1 in hepatocellular carcinoma promotes cancer growth and metastasis. Oncotarget 8(17):28683–28695PubMedPubMedCentralGoogle Scholar
  21. Liu L, Wang HJ, Meng T, Lei C, Yang XH, Wang QS, Jin B, Zhu JF (2019) lncRNA GAS5 inhibits cell migration and invasion and promotes autophagy by targeting miR-222-3p via the GAS5/PTEN-signaling pathway in CRC. Mol Ther Nucleic Acids 17:644–656CrossRefGoogle Scholar
  22. Ma C, Shi X, Zhu Q, Li Q, Liu Y, Yao Y, Song Y (2016) The growth arrest-specific transcript 5 (GAS5): a pivotal tumor suppressor long noncoding RNA in human cancers. Tumour Biol 37(2):1437–1444CrossRefGoogle Scholar
  23. McGlynn KA, London WT (2011) The global epidemiology of hepatocellular carcinoma: present and future. Clin Liver Dis 15(2):223–243CrossRefGoogle Scholar
  24. Pickard MR, Williams GT (2015) Molecular and Cellular Mechanisms of Action of Tumour Suppressor GAS5 LncRNA. Genes (Basel) 6(3):484–499CrossRefGoogle Scholar
  25. Prensner JR, Chinnaiyan AM (2011) The emergence of lncRNAs in cancer biology. Cancer Discov 1(5):391–407CrossRefGoogle Scholar
  26. Shi X, Sun M, Liu H, Yao Y, Song Y (2013) Long non-coding RNAs: a new frontier in the study of human diseases. Cancer Lett 339(2):159–166CrossRefGoogle Scholar
  27. Spizzo R, Almeida MI, Colombatti A, Calin GA (2012) Long non-coding RNAs and cancer: a new frontier of translational research? Oncogene 31(43):4577–4587CrossRefGoogle Scholar
  28. Taft RJ, Pang KC, Mercer TR, Dinger M, Mattick JS (2010) Non-coding RNAs: regulators of disease. J Pathol 220(2):126–139CrossRefGoogle Scholar
  29. Takahashi K, Yan IK, Wood J, Haga H, Patel T (2014) Involvement of extracellular vesicle long noncoding RNA (linc-VLDLR) in tumor cell responses to chemotherapy. Mol Cancer Res 12(10):1377–1387CrossRefGoogle Scholar
  30. Waghray A, Murali AR, Menon KN (2015) Hepatocellular carcinoma: from diagnosis to treatment. World J Hepatol 7(8):1020–1029CrossRefGoogle Scholar
  31. Wen Q, Liu Y, Lyu H, Xu X, Wu Q, Liu N, Yin Q, Li J, Sheng X (2017) Long noncoding RNA GAS5, which acts as a tumor suppressor via microRNA 21, regulates cisplatin resistance expression in cervical cancer. Int J Gynecol Cancer 27(6):1096–1108CrossRefGoogle Scholar
  32. Wu CY, Lin JT, Ho HJ, Su CW, Lee TY, Wang SY, Wu C, Wu JC (2014) Association of nucleos(t)ide analogue therapy with reduced risk of hepatocellular carcinoma in patients with chronic hepatitis B: a nationwide cohort study. Gastroenterology 147(1):143–151CrossRefGoogle Scholar
  33. Yoon JH, Abdelmohsen K, Gorospe M (2014) Functional interactions among microRNAs and long noncoding RNAs. Semin Cell Dev Biol 34:9–14CrossRefGoogle Scholar
  34. Yuan SX, Yang F, Yang Y, Tao QF, Zhang J, Huang G, Yang Y, Wang RY, Yang S, Huo XS, Zhang L, Wang F, Sun SH, Zhou WP (2012) Long noncoding RNA associated with microvascular invasion in hepatocellular carcinoma promotes angiogenesis and serves as a predictor for hepatocellular carcinoma patients’ poor recurrence-free survival after hepatectomy. Hepatology 56(6):2231–2241CrossRefGoogle Scholar
  35. Zhang F, Yang C, Xing Z, Liu P, Zhang B, Ma X, Huang L, Zhuang L (2019) LncRNA GAS5-mediated miR-1323 promotes tumor progression by targeting TP53INP1 in hepatocellular carcinoma. Onco Targets Ther 12:4013–4023CrossRefGoogle Scholar
  36. Zhu S, Wu H, Wu F, Nie D, Sheng S, Mo YY (2008) MicroRNA-21 targets tumor suppressor genes in invasion and metastasis. Cell Res 18(3):350–359CrossRefGoogle Scholar
  37. Zong Y, Zhang Y, Sun X, Xu T, Cheng X, Qin Y (2019) miR-221/222 promote tumor growth and suppress apoptosis by targeting lncRNA GAS5 in breast cancer. Biosci Rep. CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Hepatobiliary SurgeryFirst Hospital of Quanzhou Affiliated to Fujian Medical UniversityQuanzhouChina

Personalised recommendations