HN1L-mediated transcriptional axis AP-2γ/METTL13/TCF3-ZEB1 drives tumor growth and metastasis in hepatocellular carcinoma


Hepatocellular carcinoma (HCC) is one of the most aggressive malignancies and lacks targeted therapies. Here, we reported a novel potential therapeutic target hematological and neurological expressed 1 like (HN1L) in HCC. First, HCC tissue microarray analysis showed that HN1L was frequently up-regulated in cancer tissues than that in normal liver tissues, which significantly associated with tumor size, local invasion, distant metastases, and poor prognosis for HCC patients. Functional studies demonstrated that ectopic expression of HN1L could increase cell growth, foci formation in monolayer culture, colony formation in soft agar and tumorigenesis in nude mice. In addition, HN1L could also promote HCC metastasis by inducing epithelial-mesenchymal transition. Inversely, silencing HN1L expression with shRNA could effectively attenuate its oncogenic function. We further showed that HN1L transcriptionally up-regulated methyltransferase like 13 (METTL13) gene in an AP-2γ dependent manner, which promoted cell proliferation and metastasis by up-regulating TCF3 and ZEB1. Importantly, administration of lentivirus-mediated shRNA interfering HN1L expression could inhibit tumorigenesis and metastasis in mice. Collectively, HN1L-mediated transcriptional axis AP-2γ/METTL13/TCF3-ZEB1 promotes HCC growth and metastasis representing a promising therapeutic target in HCC treatment.

Access options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8


  1. 1.

    Ko MS, Kitchen JR, Wang X, Threat TA, Wang X, Hasegawa A, et al. Large-scale cDNA analysis reveals phased gene expression patterns during preimplantation mouse development. Development. 2000;127:1737–49.

    PubMed  Google Scholar 

  2. 2.

    Petroziello J, Yamane A, Westendorf L, Thompson M, McDonagh C, Cerveny C, et al. Suppression subtractive hybridization and expression profiling identifies a unique set of genes overexpressed in non-small-cell lung cancer. Oncogene. 2004;23:7734–45.

    CAS  Article  Google Scholar 

  3. 3.

    Li L, Zeng TT, Zhang BZ, Li Y, Zhu YH, Guan XY. Overexpression of HN1L promotes cell malignant proliferation in non-small cell lung cancer. Cancer Biol Ther. 2017;18:904–15.

    CAS  Article  Google Scholar 

  4. 4.

    Laughlin KM, Luo D, Liu C, Shaw G, Warrington KH Jr., Law BK, et al. Hematopoietic- and neurologic-expressed sequence 1 (Hn1) depletion in B16.F10 melanoma cells promotes a differentiated phenotype that includes increased melanogenesis and cell cycle arrest. Differ Res Biol Divers. 2009;78:35–44.

    CAS  Article  Google Scholar 

  5. 5.

    Goto T, Hisatomi O, Kotoura M, Tokunaga F. Induced expression of hematopoietic- and neurologic-expressed sequence 1 in retinal pigment epithelial cells during newt retina regeneration. Exp Eye Res. 2006;83:972–80.

    CAS  Article  Google Scholar 

  6. 6.

    Zhang ZG, Chen WX, Wu YH, Liang HF, Zhang BX. MiR-132 prohibits proliferation, invasion, migration, and metastasis in breast cancer by targeting HN1. Biochem Biophys Res Commun. 2014;454:109–14.

    CAS  Article  Google Scholar 

  7. 7.

    Zhang C, Xu B, Lu S, Zhao Y, Liu P. HN1 contributes to migration, invasion, and tumorigenesis of breast cancer by enhancing MYC activity. Mol Cancer. 2017;16:90.

    Article  Google Scholar 

  8. 8.

    Laughlin KM, Luo D, Liu C, Shaw G, Warrington KH Jr., Qiu J, et al. Hematopoietic- and neurologic-expressed sequence 1 expression in the murine GL261 and high-grade human gliomas. Pathol Oncol Res. 2009;15:437–44.

    CAS  Article  Google Scholar 

  9. 9.

    Varisli L, Ozturk BE, Akyuz GK, Korkmaz KS. HN1 negatively influences the beta-catenin/E-cadherin interaction, and contributes to migration in prostate cells. J Cell Biochem. 2015;116:170–8.

    CAS  Article  Google Scholar 

  10. 10.

    Jia P, Wei G, Zhou C, Gao Q, Wu Y, Sun X, et al. Upregulation of MiR-212 inhibits migration and tumorigenicity and inactivates Wnt/beta-Catenin signaling in human hepatocellular carcinoma. Technol Cancer Res Treat. 2018;17:1533034618765221.

    Article  Google Scholar 

  11. 11.

    Yang S, Kim CY, Hwang S, Kim E, Kim H, Shim H, et al. COEXPEDIA: exploring biomedical hypotheses via co-expressions associated with medical subject headings (MeSH). Nucleic Acids Res. 2017;45(D1):D389–D96.

    CAS  Article  Google Scholar 

  12. 12.

    Thiery JP, Acloque H, Huang RY, Nieto MA. Epithelial-mesenchymal transitions in development and disease. Cell. 2009;139:871–90.

    CAS  Article  Google Scholar 

  13. 13.

    Takahashi A, Tokita H, Takahashi K, Takeoka T, Murayama K, Tomotsune D, et al. A novel potent tumour promoter aberrantly overexpressed in most human cancers. Sci Rep. 2011;1:15.

    Article  Google Scholar 

  14. 14.

    Rebouissou S, Amessou M, Couchy G, Poussin K, Imbeaud S, Pilati C, et al. Frequent in-frame somatic deletions activate gp130 in inflammatory hepatocellular tumours. Nature. 2009;457:200–4.

    CAS  Article  Google Scholar 

  15. 15.

    Orchard S, Ammari M, Aranda B, Breuza L, Briganti L, Broackes-Carter F, et al. The MIntAct project–IntAct as a common curation platform for 11 molecular interaction databases. Nucleic Acids Res. 2014;42(Database issue):D358–63.

    CAS  Article  Google Scholar 

  16. 16.

    Yachdav G, Kloppmann E, Kajan L, Hecht M, Goldberg T, Hamp T, et al. Predict Protein–an open resource for online prediction of protein structural and functional features. Nucleic Acids Res. 2014;42(Web Server issue):W337–43.

    CAS  Article  Google Scholar 

  17. 17.

    Braso-Maristany F, Filosto S, Catchpole S, Marlow R, Quist J, Francesch-Domenech E, et al. PIM1 kinase regulates cell death, tumor growth and chemotherapy response in triple-negative breast cancer. Nat Med. 2016;22:1303–13.

    CAS  Article  Google Scholar 

  18. 18.

    Scheller H, Tobollik S, Kutzera A, Eder M, Unterlehberg J, Pfeil I, et al. c-Myc overexpression promotes a germinal center-like program in Burkitt's lymphoma. Oncogene. 2010;29:888–97.

    CAS  Article  Google Scholar 

  19. 19.

    Zhou G, Wang J, Zhang Y, Zhong C, Ni J, Wang L, et al. Cloning, expression and subcellular localization of HN1 and HN1L genes, as well as characterization of their orthologs, defining an evolutionarily conserved gene family. Gene. 2004;331:115–23.

    CAS  Article  Google Scholar 

  20. 20.

    Nagase T, Ishikawa K, Suyama M, Kikuno R, Hirosawa M, Miyajima N, et al. Prediction of the coding sequences of unidentified human genes. XII. The complete sequences of 100 new cDNA clones from brain which code for large proteins in vitro. DNA Res. 1998;5:355–64.

    CAS  Article  Google Scholar 

  21. 21.

    Zhang Z, Zhang G, Kong C, Zhan B, Dong X, Man X. METTL13 is downregulated in bladder carcinoma and suppresses cell proliferation, migration and invasion. Sci Rep. 2016;6:19261.

    CAS  Article  Google Scholar 

  22. 22.

    Kim J, Woo AJ, Chu J, Snow JW, Fujiwara Y, Kim CG, et al. A Myc network accounts for similarities between embryonic stem and cancer cell transcription programs. Cell. 2010;143:313–24.

    CAS  Article  Google Scholar 

  23. 23.

    Shah M, Rennoll SA, Raup-Konsavage WM, Yochum GS. A dynamic exchange of TCF3 and TCF4 transcription factors controls MYC expression in colorectal cancer cells. Cell Cycle. 2015;14:323–32.

    CAS  Article  Google Scholar 

  24. 24.

    Song XF, Chang H, Liang Q, Guo ZF, Wu JW. ZEB1 promotes prostate cancer proliferation and invasion through ERK1/2 signaling pathway. Eur Rev Med Pharmacol Sci. 2017;21:4032–8.

    PubMed  Google Scholar 

  25. 25.

    Slyper M, Shahar A, Bar-Ziv A, Granit RZ, Hamburger T, Maly B, et al. Control of breast cancer growth and initiation by the stem cell-associated transcription factor TCF3. Cancer Res. 2012;72:5613–24.

    CAS  Article  Google Scholar 

  26. 26.

    Nguyen H, Merrill BJ, Polak L, Nikolova M, Rendl M, Shaver TM, et al. Tcf3 and Tcf4 are essential for long-term homeostasis of skin epithelia. Nat Genet. 2009;41:1068–75.

    CAS  Article  Google Scholar 

  27. 27.

    Zhang P, Sun Y, Ma L. ZEB1: at the crossroads of epithelial-mesenchymal transition, metastasis and therapy resistance. Cell Cycle. 2015;14:481–7.

    CAS  Article  Google Scholar 

  28. 28.

    Ma P, Ni K, Ke J, Zhang W, Feng Y, Mao Q. miR-448 inhibits the epithelial-mesenchymal transition in breast cancer cells by directly targeting the E-cadherin repressor ZEB1/2. Exp. Biol. Med. 2018;1535370218754848.

  29. 29.

    Sinn PL, Arias AC, Brogden KA, McCray PB Jr. Lentivirus vector can be readministered to nasal epithelia without blocking immune responses. J Virol. 2008;82:10684–92.

    CAS  Article  Google Scholar 

  30. 30.

    Bie CQ, Liu XY, Cao MR, Huang QY, Tang HJ, Wang M, et al. Lentivirus-mediated RNAi knockdown of insulin-like growth factor-1 receptor inhibits the growth and invasion of hepatocellular carcinoma via down-regulating midkine expression. Oncotarget. 2016;7:79305–18.

    Article  Google Scholar 

  31. 31.

    Xie S, Wang G, Chen G, Zhu M, Lv G. Lentivirus-mediated knockdown of P27RF-Rho inhibits hepatocellular carcinoma cell growth. Contemp Oncol. 2017;21:35–41.

    CAS  Google Scholar 

  32. 32.

    Jiang L, Yan Q, Fang S, Liu M, Li Y, Yuan YF, et al. Calcium-binding protein 39 promotes hepatocellular carcinoma growth and metastasis by activating extracellular signal-regulated kinase signaling pathway. Hepatology. 2017;66:1529–45.

    CAS  Article  Google Scholar 

  33. 33.

    Seo JH, Jeong ES, Choi YK. Therapeutic effects of lentivirus-mediated shRNA targeting of cyclin D1 in human gastric cancer. BMC Cancer. 2014;14:175.

    Article  Google Scholar 

  34. 34.

    Lim HY, Sohn I, Deng S, Lee J, Jung SH, Mao M, et al. Prediction of disease-free survival in hepatocellular carcinoma by gene expression profiling. Ann Surg Oncol. 2013;20:3747–53.

    Article  Google Scholar 

  35. 35.

    Chandrashekar DS, Bashel B, Balasubramanya SAH, Creighton CJ, Ponce-Rodriguez I, Chakravarthi B, et al. UALCAN: a portal for facilitating tumor subgroup gene expression and survival analyses. Neoplasia. 2017;19:649–58.

    CAS  Article  Google Scholar 

  36. 36.

    Tang Z, Li C, Kang B, Gao G, Li C, Zhang Z. GEPIA: a web server for cancer and normal gene expression profiling and interactive analyses. Nucleic Acids Res. 2017;45(W1):W98–W102.

    CAS  Article  Google Scholar 

Download references


This work was supported by grants from the National Basic Research Program of China (2012CB967001), the China National Key Sci-Tech Special Project of Infectious Diseases (2018ZX10723204-006-005), the National Natural Science Foundation of China (81772554, 81472250, and 81472255), the China Postdoctoral Science Fund (2018M631030), the Hong Kong Research Grant Council General Research Fund (HKU/7668/11M, 767313), the Hong Kong Theme-based Research Scheme Fund (T12-704/16-R), and the Hong Kong Research Grant Council Collaborative Research Funds (C7027-14G and C7038-14G). Professor X.-Y.G. is Sophie YM Chan Professor in Cancer Research.

Author contributions:

L.L., Y.-L.Z., C.J., and S.F.: acquisition, analysis and interpretation of data; L.L.: drafting of the manuscript; T.-T.Z., Y.-H.Z., Y.L., and D.X.: technical and material support; X.-Y.G.: study design and supervision.

Author information



Corresponding author

Correspondence to Xin-Yuan Guan.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Edited by J.P. Medema

Supplementary information

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Li, L., Zheng, Y., Jiang, C. et al. HN1L-mediated transcriptional axis AP-2γ/METTL13/TCF3-ZEB1 drives tumor growth and metastasis in hepatocellular carcinoma. Cell Death Differ 26, 2268–2283 (2019).

Download citation