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Journal of Environmental Pathology, Toxicology and Oncology
Factor de Impacto: 1.241 Factor de Impacto de 5 años: 1.349 SJR: 0.519 SNIP: 0.613 CiteScore™: 1.61

ISSN Imprimir: 0731-8898
ISSN En Línea: 2162-6537

Journal of Environmental Pathology, Toxicology and Oncology

DOI: 10.1615/JEnvironPatholToxicolOncol.2019028282
pages 119-131

Evaluation of LncRNA ANRIL Potential in Hepatic Cancer Progression

Yongjian Ji
Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, 324 JingwuWfeiqi Road, Jinan, Shandong Province 250021, China
Hao Sun
Department of Gastroenterology, Dezhou People's Hospital, 1751 Xinhu Road, Dezhou, Shandong Province 253014, China
Haiqing Liang
Department of Gastroenterology, Dezhou People's Hospital, 1751 Xinhu Road, Dezhou, Shandong Province 253014, China
Yong Wang
Department of Gastroenterology, Dezhou People's Hospital, 1751 Xinhu Road, Dezhou, Shandong Province 253014, China
Meili Lu
Department of Gastroenterology, Dezhou People's Hospital, 1751 Xinhu Road, Dezhou, Shandong Province 253014, China
Zhaoyang Guo
Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, 324 JingwuWfeiqi Road, Jinan, Shandong Province 250021, China
Zhuozhen Lv
Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, 324 JingwuWfeiqi Road, Jinan, Shandong Province 250021, China
Wanhua Ren
Department of Gastroenterology, Shandong Provincial Hospital Affiliated to Shandong University, 324 JingwuWfeiqi Road, Jinan, Shandong Province 250021, China


Background/Aims: LncRNAs are significant regulators in multiple cancers including hepatocellular carcinoma (HCC). Recently, lncRNA ANRIL has been reported to be elevated during multiple cancer types, exhibiting oncogenic roles. However, the exact biological mechanism of ANRIL is still poorly understood in HCC.
Methods: Quantitative real-time polymerase chain reaction (qRT-PCR) assays were utilized to detect expressions of ANRIL, miR-384, and STAT3. CCK8 and EDU assays were employed to evaluate HCC cell proliferation. A flow cytometry assay was used to detect the HCC cell cycle and cell apoptosis. The scratch migration and Transwell invasion assays were performed to test cell migration and invasion, respectively. RIP and RNA pull-down assays were carried out to confirm the correlation between ANRIL and miR-384. The dual-luciferase reporter assay was used to prove the association between miR-384 and STAT3. Western blotting analysis was performed to examine protein levels of STAT3. IHC and HE staining were employed to detect Ki-67 and histopathology.
Results: ANRIL expression was upregulated in HCC cells, including SMCC7721, HepG2, MHCC-97H, SNU449 and HUH-7 cells, in comparison to the normal human liver cells LO2. Knockdown of ANRIL suppressed HCC cell proliferation and induced cell cycle arrest and apoptosis. HCC cell migration and invasion capacity were inhibited by inhibition of ANRIL. Bioinformatics analyses revealed that ANRIL could interact with miR-384. miR-384 was significantly decreased in HCC cells, and overexpression of miR-384 repressed HCC progression. STAT3 was predicted as a target of miR-384, and miR-384 can modulate STAT3 levels negatively in vitro. ANRIL can suppress HCC development through regulating miR-384 and STAT3 in vivo.
Conclusion: ANRIL is involved in HCC progression by direct targeting of miR-384 and STAT3. Also, ANRIL could act as a potential candidate for HCC diagnosis, prognosis, and therapy.


  1. Han K, Kim JH, Ko GY, Gwon DI, Sung KB. , Treatment of hepatocellular carcinoma with portal venous tumor thrombosis: a comprehensive review. World J Gastroenterol. 2016;22:407–16.

  2. Baghy K, Tatrai P, Regos E, Kovalszky I. , Proteoglycans in liver cancer. World J Gastroenterol. 2016;22:379–93.

  3. Faloppi L, Scartozzi M, Maccaroni E, Di Pietro Paolo M, Berardi R, Del Prete M, Cascinu S., Evolving strategies for the treatment of hepatocellular carcinoma: from clinical-guided to molecularly tailored therapeutic options. Cancer Treatment Rev. 2011;37:169–77.

  4. Rahbari NN, Mehrabi A, Mollberg NM, Muller SA, Koch M, Buchler MW, Weitz J. , Hepatocellular carcinoma: current management and perspectives for the future. Annal Surg. 2011;253:453–69.

  5. Hung T, Chang HY. , Long noncoding RNA in genome regulation: prospects and mechanisms. RNA Biol. 2010; 7: 582–85.

  6. Lee JT. , Epigenetic regulation by long noncoding RNAs. Science. 2012;338:1435–39.

  7. Cesana M, Cacchiarelli D, Legnini I, Santini T, Sthandier O, Chinappi M, Tramontano A, Bozzoni I., A long noncoding RNA controls muscle differentiation by functioning as a competing endogenous RNA. Cell. 2011;147:358–69.

  8. Li T, Liu Y, Sun Y. , Long non-coding RNA AB209630 suppresses cell proliferation and metastasis in human hepatocellular carcinoma. Exper Therap Med. 2017; 14: 3419–24.

  9. Li J, Zhang Q, Fan X, Mo W, Dai W, Feng J, Wu L, Liu T, Li S, Xu S, Wang W, Lu X, Yu Q, Chen K, Xia Y, Lu J, Zhou Y, Xu L, Guo C., The long noncoding RNA TUG1 acts as a competing endogenous RNA to regulate the hedgehog pathway by targeting miR-132 in hepatocellular carcinoma. Oncotarget. 2017;8:65932–45.

  10. Zhu L, Yang N, Chen J, Zeng T, Yan S, Liu Y, Yu G, Chen Q, Du G, Pan W, Li X, Zhou H, Huang A, Tang H., LINC00052 upregulates EPB41l3 to inhibit migration and invasion of hepatocellular carcinoma by binding miR-452-5P. Oncotarget. 2017;8:63724–37.

  11. Pasmant E, Laurendeau I, Heron D, Vidaud M, Vidaud D, Bieche I., 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. 2007;67:3963–69.

  12. Zhang EB, Kong R, Yin DD, You LH, Sun M, Han L, Xu TP, Xia R, Yang JS, De W, Chen J., Long noncoding RNA ANRIL indicates a poor prognosis of gastric cancer and promotes tumor growth by epigenetically silencing of miR-99A/miR-449A. Oncotarget. 2014;5:2276–92.

  13. Iranpour M, Soudyab M, Geranpayeh L, Mirfakhraie R, Azargashb E, Movafagh A, Ghafouri-Fard S., Expression analysis of four long noncoding RNAs in breast cancer. Tumour Biol. 2016;37:2933–40.

  14. Zhu H, Li X, Song Y, Zhang P, Xiao Y, Xing Y. , 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. 2015;467:223–28.

  15. Lujambio A, Lowe SW. , The microcosmos of cancer. Nature. 2012;482:347–55.

  16. Pencheva N, Tavazoie SF. , Control of metastatic progression by microRNA regulatory networks. Nature Cell Biol. 2013;15:546–54.

  17. Hu D, Shen D, Zhang M, Jiang N, Sun F, Yuan S, Wan K. , miR-488 suppresses cell proliferation and invasion by targeting ADAM9 and lncRNA HULC in hepatocellular carcinoma. Am J Cancer Res. 2017;7:2070–80.

  18. Yang L, Peng F, Qin J, Zhou H, Wang B. , Downregulation of microRNA-196A inhibits human liver cancer cell proliferation and invasion by targeting FOXO1. Oncol Rep. 2017; 38:2148–54.

  19. Li Q, Li S, Wu Y, Gao F. , miRNA-708 functions as a tumour suppressor in hepatocellular carcinoma by targeting SMAD3. Oncol Lett. 2017;14:2552–58.

  20. Wang YX, Chen YR, Liu SS, Ye YP, Jiao HL, Wang SY, Xiao ZY, Wei WT, Qiu JF, Liang L, Liao WT, Ding YQ., miR-384 inhibits human colorectal cancer metastasis by targeting kras and cdc42. Oncotarget. 2016; 7: 84826–38.

  21. Li Z, Yu X, Shen J. , ANRIL: a pivotal tumor suppressor long non-coding RNA in human cancers. Tumour Biol. 2016;37:5657–61.

  22. Aguilo F, Di Cecilia S, Walsh MJ. , Long non-coding RNA ANRIL and polycomb in human cancers and cardiovascular disease. Curr Topics Microbiol Immunol. 2016; 394:29–39.

  23. Peng L, Yuan X, Jiang B, Tang Z, Li GC. , lncRNAs: key players and novel insights into cervical cancer. Tumour Biol. 2016;37:2779–88.

  24. Xu R, Mao Y, Chen K, He W, Shi W, Han Y. , The long noncoding RNA ANRIL acts as an oncogene and contributes to paclitaxel resistance of lung adenocarcinoma A549 cells. Oncotarget. 2017;8:39177–84.

  25. Zou ZW, Ma C, Medoro L, Chen L, Wang B, Gupta R, Liu T, Yang XZ, Chen TT, Wang RZ, Zhang WJ, Li PD, LncRNA ANRIL is up-regulated in nasopharyngeal carcinoma and promotes the cancer progression via increasing proliferation, reprograming cell glucose metabolism and inducing side-population stem-like cancer cells. Oncotarget. 2016;7:61741–54.

  26. Zhao JJ, Hao S, Wang LL, Hu CY, Zhang S, Guo LJ, Zhang G, Gao B, Jiang Y, Tian WG, Luo DL., Long non-coding RNA ANRIL promotes the invasion and metastasis of thyroid cancer cells through TGF-beta/SMAD signaling pathway. Oncotarget. 2016; 7: 57903–18.

  27. Hua L, Wang CY, Yao KH, Chen JT, Zhang JJ, Ma WL. , High expression of long non-coding RNA ANRIL is associated with poor prognosis in hepatocellular carcinoma. Int J Clin Exper Pathol. 2015;8:3076–82.

  28. Huang MD, Chen WM, Qi FZ, Xia R, Sun M, Xu TP, Yin L, Zhang EB, De W, Shu YQ., Long non-coding RNA ANRIL is upregulated in hepatocellular carcinoma and regulates cell apoptosis by epigenetic silencing of KLF2. J Hematol Oncol. 2015;8:50.

  29. Liz J, Esteller M. , LncRNAs and microRNAs with a role in cancer development. Biochimica et Biophysica Acta. 2016;1859:169–76.

  30. Tay Y, Rinn J, Pandolfi PP. , The multilayered complexity of CERNA crosstalk and competition. Nature. 2014;505:344–52.

  31. Wang P, Fu T, Wang X, Zhu W., Primary, study of miRNA expression patterns in laryngeal carcinoma by microarray. Lin chuang er bi yan hou tou jing wai ke za zhi= J Clin Otorhinolaryngol Head Neck Surg. 2010 Jun;24(12):535–38 (in Chinese).

  32. Zheng J, Liu X, Wang P, Xue Y, Ma J, Qu C, Liu Y. , CRNDE promotes malignant progression of glioma by attenuating miR-384/PIWIL4/STAT3 axis. Mol Ther. 2016; 24:1199–215.

  33. Chen Z, Yu C, Zhan L, Pan Y, Chen L, Sun C. , LncRNA CRNDE promotes hepatic carcinoma cell proliferation, migration and invasion by suppressing miR-384. Am J Cancer Res. 2016;6:2299–309.

  34. Hu W, Jin P, Liu W. , Periostin contributes to cisplatin resistance in human non-small cell lung cancer a549 cells via activation of STAT3 and AKT and upregulation of survivin. Cell Physiol Biochem. 2016;38:1199–208.

  35. de la Iglesia N, Konopka G, Puram SV, Chan JA, Bachoo RM, You MJ, Levy DE, Depinho RA, Bonni A. , Identification of a PTEN-regulated STAT3 brain tumor suppressor pathway. Genes Develop. 2008;22:449–62.

  36. Epling-Burnette PK, Liu JH, Catlett-Falcone R, Turkson J, Oshiro M, Kothapalli R, Li Y, Wang JM, Yang-Yen HF, Karras J, Jove R, Loughran TP, Jr. , Inhibition of STAT3 signaling leads to apoptosis of leukemic large granular lymphocytes and decreased MCL-1 expression. J Clin Investig. 2001;107:351–62.

  37. Kim KW, Mutter RW, Cao C, Albert JM, Shinohara ET, Sekhar KR, Lu B., Inhibition of signal transducer and activator of transcription 3 activity results in down-regulation of survivin following irradiation. Mol Cancer Therap. 2006;5:2659–65.

  38. Wang M, Meng B, Liu Y, Yu J, Chen Q, Liu Y., miR-124 inhibits growth and enhances radiation-induced apoptosis in non-small cell lung cancer by inhibiting STAT3. Cell Physiol Biochem. 2017;44:2017–28.

  39. Tao Y, Yang S, Wu Y, Fang X, Wang Y, Song Y, Han T., MicroRNA-216a inhibits the metastasis of 412 gastric cancer cells by targeting JAK2/STAT3-mediated EMT process. Oncotarget. 2017;8:88870–81.

  40. Xue F, Liu Y, Zhang H, Wen Y, Yan L, Tang Q, Xiao E, Zhang D., Let-7a enhances the sensitivity of hepatocellular carcinoma cells to cetuximab by regulating STAT3 expression. Oncotarget Ther. 2016;9:7253–61.

  41. Sun X, Zhang J, Hou Z, Han Q, Zhang C, Tian Z, miR-146a is directly regulated by STAT3 in human hepatocellular carcinoma cells and involved in anti- tumor immune suppression. Cell Cycle. 2015;14:243–52.

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