A1CF-Axin2 signal axis regulates apoptosis and migration in Wilms tumor-derived cells through Wnt/β-catenin pathway

  • Dongsheng Ni
  • Jianing Liu
  • Yanxia Hu
  • Yamin Liu
  • Yuping Gu
  • Qin Zhou
  • Yajun XieEmail author


A1CF, a complementary factor of APOBEC-1, is involved in many cellular processes for its mRNA editing role, such as cell proliferation, apoptosis, and migration. Here, we explored the regulatory function of A1CF in Wilms tumor-derived cells. Quantitative real-time PCR was performed to detect the mRNA level of A1CF, Axin2, β-Catenin, CCND1 or NKD1 in A1CF-depleted or A1CF-overexpression G401 cells. Western bolt was used to analyze the expression of A1CF, Axin2, and β-catenin protein. The cell apoptosis and migration ability were determined using flow cytometry assay or wound healing, respectively. Our study demonstrated that overexpression of A1CF, Axin2 was upregulated and knockdown of A1CF decreased Axin2 expression at mRNA and protein levels in G401 cells. Besides, knockdown of A1CF further upregulated β-catenin, the classical regulator of Wnt signal pathway, and increased CCND1 and NKD1, the target genes of Wnt/β-catenin. Furthermore, overexpression of Axin2 partly rescued the expression of β-catenin in A1CF-deficiency stable G401 cells. In Wnt agonist BML-284 treated G401 cells, A1CF was increased like other classical regulator of Wnt signal pathway, such as Axin2 and β-catenin. Meanwhile, knockdown of Axin2 rescued β-catenin expression which was decreased in A1CF overexpression condition with BML-284. Further, overexpression of A1CF reduced cell apoptosis but promoted cell migration, and overexpression of Axin2 got similar results. In A1CF-decreased stable G401 cells, overexpression of Axin2 partly rescued the cell apoptosis and migration. We find that A1CF is a positive regulator of Axin2, a Wnt/β-catenin pathway inhibitor, and A1CF-Axin2 signal axis regulates Wilms tumor-derived cells’ apoptosis and migration through Axin2.


A1CF Axin2 Cell migration Cell apoptosis Wnt/β-catenin pathway 



The authors are grateful to all the fellows in the Division of Molecular Nephrology and the Creative Training Center for undergraduates and graduates, the M.O.E. Key Laboratory of Laboratory Medical Diagnostics, the College of Laboratory Medicine, Chongqing Medical University.

Funding information

This work is funded by the National Natural Science Foundation for Young Scientists of China (Grant No. 31701218) and the Basic Science and Frontier Technology Research Program of Chongqing Science and Technology Commission (Grant No. cstc2017jcyjA0390) by Yajun Xie Ph.D.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interests.

Supplementary material

11626_2019_335_Fig5_ESM.png (627 kb)

Supplementary figure. Western blot analysis of A1CF, Axin2 and β-Catenin expression in indicated G401 cells (A). The qRT-PCR analysis of Axin2, β-Catenin, CCND1 and NKD1 expression in Axin2-deficiency or control G401 cells (B). Data are presented as means ±standard error of the mean (SEM.) from three independent experiments. *** p < 0.001, indicate statistically significant differences. Western blot analysis of Axin1 and DKK1 expression in A1CF-deficiency or control G401 cells (C). (PNG 626 kb)

11626_2019_335_MOESM1_ESM.tif (2.2 mb)
High Resolution Image (TIF 2280 kb)


  1. Blanc V, Sessa KJ, Kennedy S, Luo J, Davidson NO (2010) Apobec-1 complementation factor modulates liver regeneration by post-transcriptional regulation of interleukin-6 mRNA stability. J Biol Chem 285:19184–19192CrossRefGoogle Scholar
  2. Clevers H, Nusse R (2012) Wnt/beta-catenin signaling and disease. Cell 149:1192–1205CrossRefGoogle Scholar
  3. Dai C, Stolz DB, Kiss LP, Monga SP, Holzman LB, Liu Y (2009) Wnt/beta-catenin signaling promotes podocyte dysfunction and albuminuria. J Am Soc Nephrol 20:1997–2008CrossRefGoogle Scholar
  4. Dome, J.S., Graf, N., Geller, J.I., Fernandez, C.V., Mullen, E.A., Spreafico, F., Van den Heuvel-Eibrink, M., and Pritchard-Jones, K. (2015). Advances in Wilms tumor treatment and biology: progress through international collaboration. J Clin Oncol 33, 2999–3007.Google Scholar
  5. Duan H, Yan Z, Chen W, Wu Y, Han J, Guo H, Qiao J (2017) TET1 inhibits EMT of ovarian cancer cells through activating Wnt/beta-catenin signaling inhibitors DKK1 and SFRP2. Gynecol Oncol 147:408–417CrossRefGoogle Scholar
  6. Figeac N, Zammit PS (2015) Coordinated action of Axin1 and Axin2 suppresses beta-catenin to regulate muscle stem cell function. Cell Signal 27:1652–1665CrossRefGoogle Scholar
  7. Fossat N, Tourle K, Radziewic T, Barratt K, Liebhold D, Studdert JB, Power M, Jones V, Loebel DA, Tam PP (2014) C to U RNA editing mediated by APOBEC1 requires RNA-binding protein RBM47. EMBO Rep 15:903–910CrossRefGoogle Scholar
  8. He W, Kang YS, Dai C, Liu Y (2011) Blockade of Wnt/beta-catenin signaling by paricalcitol ameliorates proteinuria and kidney injury. J Am Soc Nephrol 22:90–103CrossRefGoogle Scholar
  9. Huang L, Wang H, Zhou Y, Ni D, Hu Y, Long Y, Liu J, Peng R, Zhou L, Liu Z, Lyu Z, Mao Z, Hao J, Li Y, Zhou Q (2016) Apobec-1 complementation factor (A1CF) inhibits epithelial-mesenchymal transition and migration of normal rat kidney proximal tubular epithelial cells. Int J Mol Sci 17Google Scholar
  10. Hwang I, Seo EY, Ha H (2009) Wnt/beta-catenin signaling: a novel target for therapeutic intervention of fibrotic kidney disease. Arch Pharm Res 32:1653–1662CrossRefGoogle Scholar
  11. Katoh M (2018) Multi-layered prevention and treatment of chronic inflammation, organ fibrosis and cancer associated with canonical WNT/β-catenin signaling activation (review). Int J Mol Med 42:713–725Google Scholar
  12. Kawakami T, Ren S, Duffield JS (2013) Wnt signalling in kidney diseases: dual roles in renal injury and repair. J Pathol 229:221–231CrossRefGoogle Scholar
  13. Kikuchi A (1999) Modulation of Wnt signaling by Axin and Axil. Cytokine Growth Factor Rev 10:255–265CrossRefGoogle Scholar
  14. Lee E, Salic A, Kruger R, Heinrich R, Kirschner MW (2003) The roles of APC and Axin derived from experimental and theoretical analysis of the Wnt pathway. PLoS Biol 1:E10CrossRefGoogle Scholar
  15. Li S, Wang C, Liu X, Hua S, Liu X (2015) The roles of AXIN2 in tumorigenesis and epigenetic regulation. Familial Cancer 14:325–331CrossRefGoogle Scholar
  16. Liu J, Wu X, Mitchell B, Kintner C, Ding S, Schultz PG (2005) A small-molecule agonist of the Wnt signaling pathway. Angew Chem Int Ed Eng 44:1987–1990CrossRefGoogle Scholar
  17. Malogolowkin M, Cotton CA, Green DM, Breslow NE, Perlman E, Miser J, Ritchey ML, Thomas PR, Grundy PE, D'Angio GJ et al (2008) Treatment of Wilms tumor relapsing after initial treatment with vincristine, actinomycin D, and doxorubicin. A report from the National Wilms Tumor Study Group. Pediatr Blood Cancer 50:236–241CrossRefGoogle Scholar
  18. Martinez CH, Dave S, Izawa J (2010) Wilms’ tumor. Adv Exp Med Biol 685:196–209CrossRefGoogle Scholar
  19. Mehta A, Kinter MT, Sherman NE, Driscoll DM (2000) Molecular cloning of apobec-1 complementation factor, a novel RNA-binding protein involved in the editing of apolipoprotein B mRNA. Mol Cell Biol 20:1846–1854CrossRefGoogle Scholar
  20. Schweigert A, Fischer C, Mayr D, von Schweinitz D, Kappler R, Hubertus J (2016) Activation of the Wnt/beta-catenin pathway is common in Wilms tumor, but rarely through beta-catenin mutation and APC promoter methylation. Pediatr Surg Int 32:1141–1146CrossRefGoogle Scholar
  21. Snyder EM, McCarty C, Mehalow A, Svenson KL, Murray SA, Korstanje R, Braun RE (2017) APOBEC1 complementation factor (A1CF) is dispensable for C-to-U RNA editing in vivo. RNA (New York, NY) 23:457–465CrossRefGoogle Scholar
  22. Wang Z, Shao M, Liu Y (2017) Promotion of Wilms’ tumor cells migration and invasion by mono-2-ethyhexyl phthalate (MEHP) via activation of NF-kappaB signals. Chem Biol Interact 270:1–8CrossRefGoogle Scholar
  23. Wu ZQ, Brabletz T, Fearon E, Willis AL, Hu CY, Li XY, Weiss SJ (2012) Canonical Wnt suppressor, Axin2, promotes colon carcinoma oncogenic activity. Proc Natl Acad Sci U S A 109:11312–11317CrossRefGoogle Scholar
  24. Yan D, Wiesmann M, Rohan M, Chan V, Jefferson AB, Guo L, Sakamoto D, Caothien RH, Fuller JH, Reinhard C, Garcia PD, Randazzo FM, Escobedo J, Fantl WJ, Williams LT (2001) Elevated expression of axin2 and hnkd mRNA provides evidence that Wnt/beta -catenin signaling is activated in human colon tumors. Proc Natl Acad Sci U S A 98:14973–14978CrossRefGoogle Scholar
  25. Yan X, Li Q, Ni D, Xie Y, He Q, Wan Q, Liu Y, Lyu Z, Mao Z, Zhou Q (2017) Apobec-1 complementation factor regulates cell migration and apoptosis through Dickkopf1 by acting on its 3′ untranslated region in MCF7 cells. Tumour Biol 39:1010428317706218Google Scholar
  26. Zhou L, Hao J, Yuan Y, Peng R, Wang H, Ni D, Gu Y, Huang L, Mao Z, Lyu Z, du Y, Liu Z, Li Y, Ju P, Long Y, Liu J, Zhou Q (2016) EIYMNVPV motif is essential for A1CF nucleus localization and A1CF (-8aa) promotes proliferation of MDA-MB-231 cells via up-regulation of IL-6. Int J Mol Sci 17Google Scholar
  27. Zorn AM (2001) Wnt signalling: antagonistic Dickkopfs. Curr Biol 11:R592–R595CrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2019

Authors and Affiliations

  • Dongsheng Ni
    • 1
  • Jianing Liu
    • 1
  • Yanxia Hu
    • 1
  • Yamin Liu
    • 1
  • Yuping Gu
    • 1
  • Qin Zhou
    • 1
  • Yajun Xie
    • 1
    Email author
  1. 1.The Ministry of Education Key Laboratory of Laboratory Medical Diagnostics, the College of Laboratory MedicineChongqing Medical UniversityChongqingChina

Personalised recommendations