ANKHD1 promotes proliferation and invasion of non‑small‑cell lung cancer cells via regulating YAP oncoprotein expression and inactivating the Hippo pathway

Liu,Xiao‑Fang; Han,Qiang; Rong,Xue‑Zhu; Yang,Man; Han,Yu‑Chen; Yu,Juan‑Han; Lin,Xu‑Yong

doi:10.3892/ijo.2020.4994

ANKHD1 promotes proliferation and invasion of non‑small‑cell lung cancer cells via regulating YAP oncoprotein expression and inactivating the Hippo pathway

Authors:
- Xiao‑Fang Liu
- Qiang Han
- Xue‑Zhu Rong
- Man Yang
- Yu‑Chen Han
- Juan‑Han Yu
- Xu‑Yong Lin
View Affiliations

Affiliations: Department of Pathology, The First Affiliated Hospital and College of Basic Medical Sciences, China Medical University, Shenyang, Liaoning 110001, P.R. China, Department of Pathology, Shanghai Chest Hospital, Shanghai Jiaotong University, Shanghai 200030, P.R. China
Published online on: February 19, 2020 https://doi.org/10.3892/ijo.2020.4994
Pages: 1175-1185
Copyright: © Liu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The ankyrin repeat and KH domain‑containing 1 (ANKHD1) protein was recently reported to be a potential member of the Hippo signaling pathway. However, its role in human non‑small‑cell lung cancer (NSCLC) has not been extensively investigated. The aim of the present study was to examine the expression of ANKHD1 in primary human tissues and cells and determine whether it is correlated with the clinical characteristics of tumor growth. The biological functions of ANKHD1 were evaluated in vitro and in vivo. Yes‑associated protein (YAP) expression and phosphorylation induced by ANKHD1 were evaluated by western blotting and immunoprecipitation. Marked upregulation of ANKHD1 protein expression was observed in NSCLC cells and tissues, which was associated with advanced pathological tumor‑node‑metastasis stage, lymph node metastasis and poor prognosis in patients with NSCLC. ANKHD1 overexpression also promoted the proliferation and invasion of NSCLC cells. ANKHD1 upregulation inactivated Hippo signaling via increasing YAP protein levels, as well as inhibiting YAP protein phosphorylation, whereas depletion of YAP abolished the effects of ANKHD1 on cell proliferation and invasion. Therefore, ANKHD1 may play an important role in NSCLC through regulating the YAP‑dependent Hippo signaling pathway.

View Figures

View References

1	Smith RK, Carroll PM, Allard JD and Simon MA: MASK, a large ankyrin repeat and KH domain-containing protein involved in Drosophila receptor tyrosine kinase signaling. Development. 129:71–82. 2002.PubMed/NCBI
2	Poulin F, Brueschke A and Sonenberg N: Gene fusion and overlapping reading frames in the mammalian genes for 4E-BP3 and MASK. J Biol Chem. 278:52290–52297. 2003. View Article : Google Scholar : PubMed/NCBI
3	Li J, Mahajan A and Tsai MD: Ankyrin repeat: A unique motif mediating protein-protein interactions. Biochemistry. 45:15168–15178. 2006. View Article : Google Scholar : PubMed/NCBI
4	Sedgwick SG and Smerdon SJ: The ankyrin repeat: A diversity of interactions on a common structural framework. Trends Biochem Sci. 24:311–316. 1999. View Article : Google Scholar : PubMed/NCBI
5	Valverde R, Edwards L and Regan L: Structure and function of KH domains. FEBS J. 275:2712–2726. 2008. View Article : Google Scholar : PubMed/NCBI
6	Fragiadaki M and Zeidler MP: Ankyrin repeat and single KH domain 1 (ANKHD1) drives renal cancer cell proliferation via binding to and altering a subset of miRNAs. J Biol Chem. 293:9570–9579. 2018. View Article : Google Scholar : PubMed/NCBI
7	Traina F, Favaro PM, Medina Sde S, Duarte Ada S, Winnischofer SM, Costa FF and Saad ST: ANKHD1, ankyrin repeat and KH domain containing 1, is overexpressed in acute leukemias and is associated with SHP2 in K562 cells. Biochim Biophys Acta. 1762:828–834. 2006. View Article : Google Scholar : PubMed/NCBI
8	Sansores-Garcia L, Atkins M, Moya IM, Shahmoradgoli M, Tao C, Mills GB and Halder G: Mask is required for the activity of the Hippo pathway effector Yki/YAP. Curr Biol. 23:229–235. 2013. View Article : Google Scholar : PubMed/NCBI
9	Du Toit A: Cell signalling: A new Hippo pathway component. Nat Rev Mol Cell Biol. 14:1962013. View Article : Google Scholar : PubMed/NCBI
10	Zhao B, Tumaneng K and Guan KL: The Hippo pathway in organ size control, tissue regeneration and stem cell self-renewal. Nat Cell Biol. 13:877–883. 2011. View Article : Google Scholar : PubMed/NCBI
11	Bao Y, Hata Y, Ikeda M and Withanage K: Mammalian Hippo pathway: From development to cancer and beyond. J Biochem. 149:361–379. 2011. View Article : Google Scholar : PubMed/NCBI
12	Chan SW, Lim CJ, Chen L, Chong YF, Huang C, Song H and Hong W: The Hippo pathway in biological control and cancer development. J Cell Physiol. 226:928–939. 2011. View Article : Google Scholar
13	Espanel X and Sudol M: Yes-associated protein and p53-binding protein-2 interact through their WW and SH3 domains. J Biol Chem. 276:14514–14523. 2001. View Article : Google Scholar : PubMed/NCBI
14	Komuro A, Nagai M, Navin NE and Sudol M: WW domain-containing protein YAP associates with ErbB-4 and acts as a co-transcriptional activator for the carboxyl-terminal fragment of ErbB-4 that translocates to the nucleus. J Biol Chem. 278:33334–33341. 2003. View Article : Google Scholar : PubMed/NCBI
15	Lapi E, Di Agostino S, Donzelli S, Gal H, Domany E, Rechavi G, Pandolfi PP, Givol D, Strano S, Lu X and Blandino G: PML, YAP, and p73 are components of a proapoptotic autoregulatory feedback loop. Mol Cell. 32:803–814. 2008. View Article : Google Scholar : PubMed/NCBI
16	Zaidi SK, Sullivan AJ, Medina R, Ito Y, van Wijnen AJ, Stein JL, Lian JB and Stein GS: Tyrosine phosphorylation controls Runx2-mediated subnuclear targeting of YAP to repress transcription. EMBO J. 23:790–799. 2004. View Article : Google Scholar : PubMed/NCBI
17	Zhang L, Ren F, Zhang Q, Chen Y, Wang B and Jiang J: The TEAD/TEF family of transcription factor Scalloped mediates Hippo signaling in organ size control. Dev Cell. 14:377–387. 2008. View Article : Google Scholar : PubMed/NCBI
18	Yu FX, Zhao B, Panupinthu N, Jewell JL, Lian I, Wang LH, Zhao J, Yuan H, Tumaneng K, Li H, et al: Regulation of the Hippo-YAP pathway by G-protein-coupled receptor signaling. Cell. 150:780–791. 2012. View Article : Google Scholar : PubMed/NCBI
19	Enderle L and McNeill H: Hippo gains weight: Added insights and complexity to pathway control. Sci Signal. 6:re72013. View Article : Google Scholar : PubMed/NCBI
20	Machado-Neto JA, Lazarini M, Favaro P, Franchi GC Jr, Nowill AE, Saad ST and Traina F: ANKHD1, a novel component of the Hippo signaling pathway, promotes YAP1 activation and cell cycle progression in prostate cancer cells. Exp Cell Res. 324:137–145. 2014. View Article : Google Scholar : PubMed/NCBI
21	Travis WD, Brambilla E, Nicholson AG, Yatabe Y, Austin JHM, Beasley MB, Chirieac LR, Dacic S, Duhig E, Flieder DB, et al: The 2015 World Health Organization Classification of Lung Tumors: Impact of genetic, clinical and radiologic advances since the 2004 classification. J Thorac Oncol. 10:1243–1260. 2015. View Article : Google Scholar : PubMed/NCBI
22	Goldstraw P: Updated staging system for lung cancer. Surg Oncol Clin N Am. 20:655–666. 2011. View Article : Google Scholar : PubMed/NCBI
23	Nishioka N, Inoue K, Adachi K, Kiyonari H, Ota M, Ralston A, Yabuta N, Hirahara S, Stephenson RO, Ogonuki N, et al: The Hippo signaling pathway components Lats and Yap pattern Tead4 activity to distinguish mouse trophectoderm from inner cell mass. Dev Cell. 16:398–410. 2009. View Article : Google Scholar : PubMed/NCBI
24	Xin M, Kim Y, Sutherland LB, Murakami M, Qi X, McAnally J, Porrello ER, Mahmoud AI, Tan W, Shelton JM, et al: Hippo pathway effector Yap promotes cardiac regeneration. Proc Natl Acad Sci USA. 110:13839–13844. 2013. View Article : Google Scholar : PubMed/NCBI
25	Chang Y, Fu XR, Cui M, Li WM, Zhang L, Li X, Li L, Sun ZC, Zhang XD, Li ZM, et al: Activated hippo signal pathway inhibits cell proliferation and promotes apoptosis in NK/T cell lymphoma cells. Cancer Med. 8:3892–3904. 2019.PubMed/NCBI
26	Dhyani A, Duarte AS, Machado-Neto JA, Favaro P, Ortega MM and Olalla Saad ST: ANKHD1 regulates cell cycle progression and proliferation in multiple myeloma cells. FEBS Lett. 586:4311–4318. 2012. View Article : Google Scholar : PubMed/NCBI
27	Dhyani A, Machado-Neto JA, Favaro P and Saad ST: ANKHD1 represses p21 (WAF1/CIP1) promoter and promotes multiple myeloma cell growth. Eur J Cancer. 51:252–259. 2015. View Article : Google Scholar
28	Machado-Neto JA, Lazarini M, Favaro P, de Melo Campos P, Scopim-Ribeiro R, Franchi Junior GC, Nowill AE, Lima PR, Costa FF, Benichou S, et al: ANKHD1 silencing inhibits Stathmin 1 activity, cell proliferation and migration of leukemia cells. Biochim Biophys Acta. 1853:583–593. 2015. View Article : Google Scholar
29	Mosavi LK, Cammett TJ, Desrosiers DC and Peng ZY: The ankyrin repeat as molecular architecture for protein recognition. Protein Sci. 13:1435–1448. 2004. View Article : Google Scholar : PubMed/NCBI
30	Sidor CM, Brain R and Thompson BJ: Mask proteins are cofactors of Yorkie/YAP in the Hippo pathway. Curr Biol. 23:223–228. 2013. View Article : Google Scholar : PubMed/NCBI
31	Zhao B, Ye X, Yu J, Li L, Li W, Li S, Yu J, Lin JD, Wang CY, Chinnaiyan AM, et al: TEAD mediates YAP-dependent gene induction and growth control. Genes Dev. 22:1962–1971. 2008. View Article : Google Scholar : PubMed/NCBI
32	Hao Y, Chun A, Cheung K, Rashidi B and Yang X: Tumor suppressor LATS1 is a negative regulator of oncogene YAP. J Biol Chem. 283:5496–5509. 2008. View Article : Google Scholar
33	Oka T, Mazack V and Sudol M: Mst2 and Lats kinases regulate apoptotic function of Yes kinase-associated protein (YAP). J Biol Chem. 283:27534–27546. 2008. View Article : Google Scholar : PubMed/NCBI
34	Remue E: TAZ interacts with zonula occludens-1 and -2 proteins in a PDZ-1 dependent manner. FEBS Lett. 584:175–180. 2010. View Article : Google Scholar
35	Ren F, Zhang L and Jiang J: Hippo signaling regulates Yorkie nuclear localization and activity through 14-3-3 dependent and independent mechanisms. Dev Biol. 337:303–312. 2010. View Article : Google Scholar :
36	Wu S, Liu Y, Zheng Y, Dong J and Pan D: The TEAD/TEF family protein Scalloped mediates transcriptional output of the Hippo growth-regulatory pathway. Dev Cell. 14:388–398. 2008. View Article : Google Scholar : PubMed/NCBI
37	Zhao B, Li L, Tumaneng K, Wang CY and Guan KL: A coordinated phosphorylation by Lats and CK1 regulates YAP stability through SCF(beta-TRCP). Genes Dev. 24:72–85. 2010. View Article : Google Scholar : PubMed/NCBI
38	Cottini F, Hideshima T, Xu C, Sattler M, Dori M, Agnelli L, ten Hacken E, Bertilaccio MT, Antonini E, Neri A, et al: Rescue of Hippo coactivator YAP1 triggers DNA damage-induced apop-tosis in hematological cancers. Nat Med. 20:599–606. 2014. View Article : Google Scholar : PubMed/NCBI
39	Machado-Neto JA, de Melo Campos P, Olalla Saad ST and Traina F: YAP1 expression in myelodysplastic syndromes and acute leukemias. Leuk Lymphoma. 55:2413–2415. 2014. View Article : Google Scholar : PubMed/NCBI