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Knockdown of LncRNA ANRIL suppresses cell proliferation, metastasis, and invasion via regulating miR-122-5p expression in hepatocellular carcinoma

  • Original Article – Cancer Research
  • Published:
Journal of Cancer Research and Clinical Oncology Aims and scope Submit manuscript

Abstract

Objective

Previous studies reported that lncRNA antisense non-coding RNA in the INK4 locus (ANRIL) was upregulated in hepatocellular carcinoma (HCC) tissues and decreased expression of ANRIL could suppress cell proliferation, metastasis, and invasion and induce apoptosis of HCC cells. However, the molecular mechanism of ANRIL involved in HCC tumorigenesis is still unknown.

Methods

The expressions of ANRIL and miR-122-5p in HCC tissues and cells were quantified by qRT-PCR. MTT assay, colony formation assay, wound healing assay, and transwell invasion assay were performed to evaluate cell growth, metastasis, and invasion, respectively. RNA immunoprecipitation (RIP) assay and luciferase reporter assay were performed to determine whether ANRIL could directly bind to miR-122-5p in HCC cells. Xenograft tumor experiment was conducted to confirm the biological role and underlying mechanism of ANRIL in vivo.

Results

The results showed that ANRIL was upregulated and miR-122-5p was downregulated in HCC tissues and cells. ANRIL was negatively correlated with miR-122-5p expression in HCC tissues. Knockdown of ANRIL or miR-122-5p overexpression suppressed HCC cell viability, colony formation ability, metastasis, and invasion. ANRIL was demonstrated to directly bind to miR-122-5p and inhibit its expression. Forced expression of ANRIL abolished the inhibitory effect of miR-122-5p overexpression on HCC progression. In vivo experiment demonstrated that ANRIL knockdown impeded tumor growth in vivo and increased miR-122-5p expression.

Conclusion

Our finding suggested that knockdown of ANRIL suppressed cell proliferation, metastasis and invasion via regulating miR-122-5p expression in HCC, illustrating the underlying mechanism of the oncogenic role of ANRIL in HCC.

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References

  • Cai H et al (2017) Long non-coding RNA taurine upregulated 1 enhances tumor-induced angiogenesis through inhibiting microRNA-299 in human glioblastoma. Oncogene 36:318

    Article  CAS  PubMed  Google Scholar 

  • Cesana M et al (2011) A long noncoding RNA controls muscle differentiation by functioning as a competing endogenous. RNA Cell 147:358

    Article  CAS  PubMed  Google Scholar 

  • Cho WC (2010) MicroRNAs: potential biomarkers for cancer diagnosis, prognosis and targets for therapy. Int J Biochem Cell Biol 42:1273–1281

    Article  CAS  PubMed  Google Scholar 

  • Cui M, Zheng M, Sun B, Wang Y, Ye L, Zhang X (2015) A long noncoding RNA perturbs the circadian rhythm of hepatoma cells to facilitate hepatocarcinogenesis 1. Neoplasia 17(2):79–88

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Eltawansy S, Gomez J, Liss K, Nivera N, Babyatsky M (2015) Syndrome of inappropriate anti-diuretic hormone secondary to non-cirrhotic primary hepatocellular carcinoma. Am J Case Rep 16:31–36

    Article  PubMed  PubMed Central  Google Scholar 

  • Ergun S et al (2015) The association of the expression of miR-122-5p and its target ADAM10 with human breast cancer. Mol Biol Rep 42:497–505. https://doi.org/10.1007/s11033-014-3793-2

    Article  CAS  PubMed  Google Scholar 

  • Gao Y et al (2015) LncRNA-HOST2 regulates cell biological behaviors in epithelial ovarian cancer through a mechanism involving microRNA let-7b. Hum Mol Genet 24:841

    Article  CAS  PubMed  Google Scholar 

  • Hua L, Wang CY, Yao KH, Chen JT, Zhang JJ, Ma WL (2015) High expression of long non-coding RNA ANRIL is associated with poor prognosis in hepatocellular carcinoma. Int J Clin Exp Pathol 8:3076–3082

    CAS  PubMed  PubMed Central  Google Scholar 

  • Huang M et al (2015a) Long non-coding RNA ANRIL is upregulated in hepatocellular carcinoma and regulates cell apoptosis by epigenetic silencing of KLF2. J Hematol Oncol 8:57

    Article  PubMed  PubMed Central  Google Scholar 

  • Huang MD et al (2015b) Long non-coding RNA ANRIL is upregulated in hepatocellular carcinoma and regulates cell apoptosis by epigenetic silencing of KLF2. J Hematol Oncol 8:50. https://doi.org/10.1186/s13045-015-0146-0

    Article  PubMed  Google Scholar 

  • Krol J, Loedige I, Filipowicz W (2010) The widespread regulation of microRNA biogenesis, function and decay. Nat Rev Genet 11:597–610

    Article  CAS  PubMed  Google Scholar 

  • Kung JTY, Colognori D, Lee JT (2013) Long noncoding RNAs: past, present, and future. Genetics 193:651

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Li G et al (2014) Long noncoding RNA plays a key role in metastasis and prognosis of hepatocellular carcinoma. Biomed Res Int 2014:780521

    PubMed  PubMed Central  Google Scholar 

  • Li T, Meng XL, Yang WQ (2017) Long noncoding RNA PVT1 acts as a “Sponge” to inhibit microRNA-152 in Gastric Cancer Cells. Digest Dis Sci. https://doi.org/10.1007/s10620-017-4508-z

    Google Scholar 

  • Liu RT, Cao JL, Yan CQ, Wang Y, An CJ, Lv HT (2017) Effects of LncRNA-HOST2 on cell proliferation, migration, invasion and apoptosis of human hepatocellular carcinoma cell line SMMC-7721. Biosci Rep

  • Lu W et al (2015) miR-122 inhibits cancer cell malignancy by targeting PKM2 in gallbladder carcinoma. Tumour Biol 37:15615–15625. https://doi.org/10.1007/s13277-015-4308-z

    Article  Google Scholar 

  • Lu Y, Zhou X, Xu L, Rong C, Shen C, Bian W (2016) Long noncoding RNA ANRIL could be transactivated by c-Myc and promote tumor progression of non-small-cell lung cancer. OncoTarg Ther 9:3077–3084. https://doi.org/10.2147/ott.s102658

    Google Scholar 

  • Ma MZ, Chu BF, Zhang Y, Weng MZ, Qin YY, Gong W, Quan ZW (2015) Long non-coding RNA CCAT1 promotes gallbladder cancer development via negative modulation of miRNA-218-5p. Cell Death Dis 6:e1583

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ma F, Wang SH, Cai Q, Jin LY, Zhou D, Ding J, Quan ZW (2017) Long non-coding RNA TUG1 promotes cell proliferation and metastasis by negatively regulating miR-300 in gallbladder carcinoma. Biomed Pharmacother = Biomedecine pharmacotherapie 88:863–869. https://doi.org/10.1016/j.biopha.2017.01.150

    Article  CAS  PubMed  Google Scholar 

  • Mang Y et al (2017) Long noncoding RNA NEAT1 promotes cell proliferation and invasion by regulating hnRNP A2 expression in hepatocellular carcinoma cells. Oncotarg Ther 10:1003

    Article  Google Scholar 

  • Mathieu EL, Belhocine M, Dao LT, Puthier D, Spicuglia S (2014) Functions of lncRNA in development and diseases. Med Sci M/s 30:790

    Google Scholar 

  • Mercer TR, Dinger ME, Mattick JS (2009) Long non-coding RNAs: insights into functions. Nat Rev Genet 10:155

    Article  CAS  PubMed  Google Scholar 

  • Naemura M, Murasaki C, Inoue Y, Okamoto H, Kotake Y (2015) Long noncoding RNA ANRIL regulates proliferation of non-small cell lung cancer and cervical cancer. Cells Anticancer Res 35:5377

    CAS  PubMed  Google Scholar 

  • Nagano T, Fraser P (2011) No-nonsense functions for long noncoding RNAs. Cell 145:178–181

    Article  CAS  PubMed  Google Scholar 

  • Nie FQ et al (2015) Long noncoding RNA ANRIL promotes non-small cell lung cancer cell proliferation and inhibits apoptosis by silencing KLF2 and P21 expression. Mol Cancer Therapeut 14:268–277. https://doi.org/10.1158/1535-7163.mct-14-0492

    Article  CAS  Google Scholar 

  • Pasmant E, Laurendeau I, Heron D, Vidaud M, Vidaud D, Bieche I (2007) Characterization of a germ-line deletion, including the entire INK4/ARF locus, in a melanoma-neural system tumor family: identification of ANRIL, an antisense noncoding RNA whose expression coclusters with. ARF Cancer Res 67:3963–3969. https://doi.org/10.1158/0008-5472.can-06-2004

    Article  CAS  PubMed  Google Scholar 

  • Qiu JJ, Wang Y, Liu YL, Zhang Y, Ding JX, Hua KQ (2016) The long non-coding RNA ANRIL promotes proliferation and cell cycle progression and inhibits apoptosis and senescence in epithelial ovarian cancer. Oncotarget 7:32478–32492. https://doi.org/10.18632/oncotarget.8744

    PubMed  PubMed Central  Google Scholar 

  • Salmena L, Poliseno L, Tay Y, Kats L, Pandolfi PP (2011) ceRNA hypothesis: the Rosetta stone of a hidden RNA language? Cell 146:353–358

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Serviss JT, Johnsson P, Dan G (2014) An emerging role for long non-coding RNAs in cancer metastasis. Front Genet 5:234

    Article  PubMed  PubMed Central  Google Scholar 

  • Sun Y, Zheng ZP, Li H, Zhang HQ, Ma FQ (2016) ANRIL is associated with the survival rate of patients with colorectal cancer, and affects cell migration and invasion in vitro. Mol Med Rep 14:1714–1720. https://doi.org/10.3892/mmr.2016.5409

    Article  CAS  PubMed  Google Scholar 

  • Tan TC et al (2010) Combined resection and radiofrequency ablation for multifocal hepatocellular carcinoma: prognosis and outcomes. WJG 16:3056

    Article  Google Scholar 

  • Tan J, Qiu K, Li M, Liang Y (2015) Double-negative feedback loop between long non-coding RNA TUG1 and miR-145 promotes epithelial to mesenchymal transition and radioresistance in human bladder cancer cells. Febs Lett 589:3175

    Article  CAS  PubMed  Google Scholar 

  • Tian L, Lei C, Long W, Yuan Y (2016) Downregulation of ZEB2-AS1 decreased tumor growth and metastasis in hepatocellular carcinoma. Mol Med Rep 14:4606–4612

    Article  Google Scholar 

  • van Meerloo J, Kaspers GJ, Cloos J (2011) Cell sensitivity assays: the MTT assay. Methods Mol Biol (Clifton NJ) 731:237–245. https://doi.org/10.1007/978-1-61779-080-5_20

    Article  Google Scholar 

  • Wang Y et al (2013) Endogenous miRNA sponge lincRNA-RoR regulates Oct4, Nanog, and Sox2 in Human embryonic stem cell self-renewal. Dev Cell 25:69

    Article  CAS  PubMed  Google Scholar 

  • Wang P, Liu YH, Yao YL, Li Z, Li ZQ, Ma J, Xue YX (2015) Long non-coding RNA CASC2 suppresses malignancy in human gliomas by miR-21. Cell Signal 27:275–282. https://doi.org/10.1016/j.cellsig.2014.11.011

    Article  CAS  PubMed  Google Scholar 

  • Wang N, Wang Q, Shen D, Sun X, Cao X, Wu D (2016) Downregulation of microRNA-122 promotes proliferation, migration, and invasion of human hepatocellular carcinoma cells by activating epithelial-mesenchymal transition. OncoTarg Ther 9:2035–2047. https://doi.org/10.2147/ott.s92378

    Article  Google Scholar 

  • Xiao JN et al. (2017) Long non-coding RNA UCA1 regulates the expression of Snail2 by miR-203 to promote hepatocellular carcinoma progression. J Cancer Res Clin Oncol:1–10

  • Xie H, Ma H, Zhou D (2013) Plasma HULC as a promising novel biomarker for the detection of hepatocellular carcinoma. Biomed Res Int 2013:1–5. https://doi.org/10.1155/2013/136106

    Google Scholar 

  • Xu Y et al (2017) Long non-coding RNA CCAT2 is associated with poor prognosis in hepatocellular carcinoma and promotes tumor metastasis by regulating Snail2-mediated epithelial-mesenchymal transition. Oncotarg Ther 10:1191

    Article  Google Scholar 

  • Yang D et al (2016) LncRNA mediated regulation of aging pathways in Drosophila melanogaster during dietary restriction. Aging 8:2182–2197

    Article  PubMed  PubMed Central  Google Scholar 

  • Yao S, Tian C, Ding Y, Ye Q, Gao Y, Yang N, Li Q (2016) Down-regulation of Krüppel-like factor-4 by microRNA-135a-5p promotes proliferation and metastasis in hepatocellular carcinoma by transforming growth factor-β1. Oncotarget 7:42566–42578

    PubMed  PubMed Central  Google Scholar 

  • Yap KL et al. (2010) Molecular interplay of the noncoding RNA ANRIL and methylated histone H3 lysine 27 by polycomb CBX7 in transcriptional silencing of INK4a. Mol Cell 38:662

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Yuan et al (2014) A long noncoding RNA activated by TGF-β promotes the invasion-metastasis cascade in hepatocellular. Carcinoma Cancer Cell 25:666

    Article  CAS  PubMed  Google Scholar 

  • Zhao N et al (2016) Expression of microRNA-195 is transactivated by Sp1 but inhibited by histone deacetylase 3 in hepatocellular carcinoma cells. BBA Gene Regul Mech 1859:933–942

    CAS  Google Scholar 

  • Zhu H, Li X, Song Y, Zhang P, Xiao Y, Xing Y (2015a) Long non-coding RNA ANRIL is up-regulated in bladder cancer and regulates bladder cancer cell proliferation and apoptosis through the intrinsic pathway. Biochem Biophys Res Commun 467:223–228. https://doi.org/10.1016/j.bbrc.2015.10.002

    Article  CAS  PubMed  Google Scholar 

  • Zhu H, Li X, Song Y, Zhang P, Xiao Y, Xing Y (2015b) Long non-coding RNA ANRIL is up-regulated in bladder cancer and regulates bladder cancer cell proliferation and apoptosis through the intrinsic pathway. Biochem Biophys Res Commun 467:223–228

    Article  CAS  PubMed  Google Scholar 

  • Zhuang LK et al (2016) MicroRNA-92b promotes hepatocellular carcinoma progression by targeting Smad7 and is mediated by long non-coding RNA. XIST Cell Death Dis 7:e2203

    Article  CAS  PubMed  Google Scholar 

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Correspondence to Xinwei Han.

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

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Ma, J., Li, T., Han, X. et al. Knockdown of LncRNA ANRIL suppresses cell proliferation, metastasis, and invasion via regulating miR-122-5p expression in hepatocellular carcinoma. J Cancer Res Clin Oncol 144, 205–214 (2018). https://doi.org/10.1007/s00432-017-2543-y

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  • DOI: https://doi.org/10.1007/s00432-017-2543-y

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