Loss of 4.1N in epithelial ovarian cancer results in EMT and matrix-detached cell death resistance
Received date: 12 Dec 2019
Accepted date: 23 Mar 2020
Published date: 15 Feb 2021
Copyright
Epithelial ovarian cancer (EOC) is one of the leading causes of death from gynecologic cancers and peritoneal dissemination is the major cause of death in patients with EOC. Although the loss of 4.1N is associated with increased risk of malignancy, its association with EOC remains unclear. To explore the underlying mechanism of the loss of 4.1N in constitutive activation of epithelial-mesenchymal transition (EMT) and matrixdetached cell death resistance, we investigated samples from 268 formalin-fixed EOC tissues and performed various in vitro and in vivo assays. We report that the loss of 4.1N correlated with progress in clinical stage, as well as poor survival in EOC patients. The loss of 4.1N induces EMT in adherent EOC cells and its expression inhibits anoikis resistance and EMT by directly binding and accelerating the degradation of 14-3-3 in suspension EOC cells. Furthermore, the loss of 4.1N could increase the rate of entosis, which aggravates cell death resistance in suspension EOC cells. Moreover, xenograft tumors in nude mice also show that the loss of 4.1N can aggravate peritoneal dissemination of EOC cells. Single-agent and combination therapy with a ROCK inhibitor and a 14-3-3 antagonist can reduce tumor spread to varying degrees. Our results not only define the vital role of 4.1N loss in inducing EMT, anoikis resistance, and entosis-induced cell death resistance in EOC, but also suggest that individual or combined application of 4.1N, 14-3-3 antagonists, and entosis inhibitors may be a promising therapeutic approach for the treatment of EOC.
Key words: epithelial ovarian cancer; 4.1N; EMT; anoikis; entosis
Dandan Wang , Letian Zhang , Ajin Hu , Yuxiang Wang , Yan Liu , Jing Yang , Ningning Du , Xiuli An , Congying Wu , Congrong Liu . Loss of 4.1N in epithelial ovarian cancer results in EMT and matrix-detached cell death resistance[J]. Protein & Cell, 2021 , 12(2) : 107 -127 . DOI: 10.1007/s13238-020-00723-9
1 |
Armstrong DK, Alvarez RD, Bakkum-Gamez JN, Barroilhet L, Behbakht K, Berchuck A, Berek JS, Chen LM, Cristea M, DeRosa M
|
2 |
Baines AJ, Lu HC, Bennett PM (2014) The protein 4.1 family: hub proteins in animals for organizing membrane proteins. Biochim Biophys Acta 1838(2):605–619
|
3 |
Bast RC Jr,Hennessy B, Mills GB (2009) The biology of ovarian cancer: new opportunities for translation. Nat Rev Cancer 9 (6):415–428
|
4 |
Bonnans C, Chou J, Werb Z (2014) Remodelling the extracellular matrix in development and disease. Nat Rev Mol Cell Biol 15 (12):786–801
|
5 |
Brooks PC, Montgomery AM, Rosenfeld M, Reisfeld RA, Hu T, Klier G, Cheresh DA (1994) Integrin alpha v beta 3 antagonists promote tumor regression by inducing apoptosis of angiogenic blood vessels. Cell 79(7):1157–1164
|
6 |
Calinisan V, Gravem D, Chen RP, Brittin S, Mohandas N, Lecomte MC, Gascard P(2006) New insights into potential functions for the protein 4.1 superfamily of proteins in kidney epithelium. Front Biosci 11:1646–1666
|
7 |
Chishti AH, Kim AC, Marfatia SM, Lutchman M, Hanspal M, Jindal H,Liu SC, Low PS, Rouleau GA, Mohandas N
|
8 |
Frame MC, Patel H, Serrels B, Lietha D, Eck MJ (2010) The FERM domain: organizing the structure and function of FAK. Nat Rev Mol Cell Biol 11(11):802–814
|
9 |
Friedl P, Alexander S (2011) Cancer invasion and the microenvironment: plasticity and reciprocity. Cell 147(5):992–1009
|
10 |
Frisch SM, Francis H (1994) Disruption of epithelial cell-matrix interactions induces apoptosis. J Cell Biol 124(4):619–626
|
11 |
Frisch SM, Schaller M,Cieply B (2013) Mechanisms that link the oncogenic epithelial-mesenchymal transition to suppression of anoikis. J Cell Sci 126(Pt 1):21–29
|
12 |
Frisch SM, Screaton RA (2001) Anoikis mechanisms. Curr Opin Cell Biol 13(5):555–562
|
13 |
Giancotti FG (2000) Complexity and specificity of integrin signalling. Nat Cell Biol 2(1):E13–E14
|
14 |
Hamann JC, Surcel A, Chen R, Teragawa C, Albeck JG, Robinson DN, Overholtzer M (2017) Entosis is induced by glucose starvation. Cell Rep 20(1):201–210
|
15 |
Hermeking H (2003) The 14-3-3 cancer connection. Nat Rev Cancer 3(12):931–943
|
16 |
Hou Z, Peng H, White DE, Wang P, Lieberman PM, Halazonetis T, Rauscher FJ 3rd (2010) 14-3-3 binding sites in the snail protein are essential for snail-mediated transcriptional repression and epithelial-mesenchymal differentiation. Cancer Res 70(11):4385–4393
|
17 |
Howlett AR, Bailey N, Damsky C, Petersen OW, Bissell MJ (1995) Cellular growth and survival are mediated by beta 1 integrins in normal human breast epithelium but not in breast carcinoma. J Cell Sci 108(Pt 5):1945–1957
|
18 |
Janssen A, Medema RH (2011) Entosis: aneuploidy by invasion. Nature Cell Biol 13(3):199–201
|
19 |
Ji Z, Shi X, Liu X,Shi Y, Zhou Q, Liu X, Li L, Ji X, Gao Y, Qi Y
|
20 |
Kipps E, Tan DS, Kaye SB (2013) Meeting the challenge of ascites in ovarian cancer: new avenues for therapy and research. Nat Rev Cancer 13(4):273–282
|
21 |
Krauss SW, Chen C, Penman S, Heald R (2003) Nuclear actin and protein 4.1: essential interactions during nuclear assembly in vitro. Proc Natl Acad Sci USA 100(19):10752–10757
|
22 |
Kroemer G, Perfettini JL (2014) Entosis, a key player in cancer cell competition. Cell Res 24(11):1280–1281
|
23 |
Mackay HL, Muller PAJ (2019) Biological relevance of cell-in-cell in cancers. Biochem Soc Trans 47(2):725–732
|
24 |
Martins I, Raza SQ, Voisin L, Dakhli H, Allouch A, Law F, Sabino D, De Jong D, Thoreau M, Mintet E
|
25 |
Martins I,Raza SQ, Voisin L, Dakhli H, Law F, De Jong D, Allouch A, Thoreau M, Brenner C, Deutsch E
|
26 |
Meredith JE Jr, Fazeli B, Schwartz MA (1993) The extracellular matrix as a cell survival factor. Mol Biol Cell 4(9):953–961
|
27 |
Nomura M,Shimizu S, Sugiyama T, Narita M, Ito T, Matsuda H, Tsujimoto Y (2015) 14-3-3 interacts directly with and negatively regulates pro-apoptotic Bax. J Biol Chem 290(11):6753
|
28 |
Overholtzer M, Mailleux AA, Mouneimne G, Normand G, Schnitt SJ, King RW, Cibas ES, Brugge JS (2007) A nonapoptotic cell death process, entosis, that occurs by cell-in-cell invasion. Cell 131 (5):966–979
|
29 |
Paoli P, Giannoni E, Chiarugi P (2013) Anoikis molecular pathways and its role in cancer progression. Biochim Biophys Acta 1833 (12):3481–3498
|
30 |
Samuel T, Weber HO, Rauch P, Verdoodt B, Eppel JT, McShea A, Hermeking H, Funk JO (2001) The G2/M regulator 14-3-3sigma prevents apoptosis through sequestration of Bax. J Biol Chem 276(48):45201–45206
|
31 |
Siegel RL, Miller KD, Jemal A (2019) Cancer statistics, 2019. CA Cancer J Clin 69(1):7–34
|
32 |
Simpson CD, Anyiwe K, Schimmer AD (2008) Anoikis resistance and tumor metastasis. Cancer Lett 272(2):177–185
|
33 |
Sun Q, Luo T, Ren Y, Florey O, Shirasawa S, Sasazuki T, Robinson DN, Overholtzer M (2014) Competition between human cells by entosis. Cell Res 24(11):1299–1310
|
34 |
Taddei ML, Giannoni E, Fiaschi T, Chiarugi P (2012) Anoikis: an emerging hallmark in health and diseases. J Pathol 226(2):380–393
|
35 |
Tan Y, Demeter MR, Ruan H, Comb MJ (2000) BAD Ser-155 phosphorylation regulates BAD/Bcl-XL interaction and cell survival. J Biol Chem 275(33):25865–25869
|
36 |
Taylor-Harris PM, Felkin LE, Birks EJ, Franklin RC, Yacoub MH, Baines AJ, Barton PJ, Pinder JC (2005) Expression of human membrane skeleton protein genes for protein 4.1 and betaIISigma2- spectrin assayed by real-time RT-PCR. Cell Mol Biol Lett 10 (1):135–149
|
37 |
Thiery JP, Acloque H, Huang RY, Nieto MA (2009) Epithelialmesenchymal transitions in development and disease. Cell 139 (5):871–890
|
38 |
Tong Y, Yang H, Xu X, Ruan J, Liang M, Wu J, Luo BJCS (2017) Effect of a hypoxic microenvironment after radiofrequency ablation on residual hepatocellular cell migration and invasion. Cancer Sci 108(4):753–762
|
39 |
Walensky LD, Blackshaw S, Liao D, Watkins CC, Weier HU, Parra M, Huganir RL, Conboy JG, Mohandas N, Snyder SH (1999) A novel neuron-enriched homolog of the erythrocyte membrane cytoskeletal protein 4.1. J Neurosci 19(15):6457–6467
|
40 |
Wang Z, Ma B, Li H, Xiao X, Zhou W, Liu F, Zhang B, Zhu M, Yang Q, Zeng Y
|
41 |
Wilker E, Yaffe MB (2004) 14-3-3 Proteins—a focus on cancer and human disease. J Mol Cell Cardiol 37(3):633–642
|
42 |
Xi C, Ren C, Hu A, Lin J, Yao Q, Wang Y, Gao Z, An X, Liu C (2013) Defective expression of protein 4.1N is correlated to tumor progression, aggressive behaviors and chemotherapy resistance in epithelial ovarian cancer. Gynecol Oncol 131(3):764–771
|
43 |
Yang Q,Zhu M, Wang Z, Li H, Zhou W, Xiao X, Zhang B, Hu W, Liu J (2016) 4.1N is involved in a flotillin-1/beta-catenin/Wnt pathway and suppresses cell proliferation and migration in non-small cell lung cancer cell lines. Tumour Biol 37(9):12713–12723
|
44 |
Zhang L, Hu A, Li M, Zhang H, Ren C, An X, Liu C (2016) 4.1N suppresses hypoxia-induced epithelial-mesenchymal transition in epithelial ovarian cancer cells. Mol Med Rep 13(1):837–844
|
45 |
Zhang Z, Vuori K, Reed JC, Ruoslahti E (1995) The alpha 5 beta 1 integrin supports survival of cells on fibronectin and up-regulates Bcl-2 expression. Proc Natl Acad Sci USA 92(13):6161–6165
|
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