Open Access

Role of TGF‑β in the motility of ShcD‑overexpressing 293 cells

  • Authors:
    • Sara Amer
    • Fadi Alsayegh
    • Zeina Mashaal
    • Salma Mohamed
    • Nour Shawa
    • Keerthi Rajan
    • Samrein B.M. Ahmed
  • View Affiliations

  • Published online on: July 23, 2019     https://doi.org/10.3892/mmr.2019.10517
  • Pages: 2667-2674
  • Copyright: © Amer 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: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The newly identified Src homology and collagen (Shc) family member ShcD was observed to be upregulated in 50% of vertical growth phase and metastatic melanomas. The aim of the present study was to investigate the mechanism by which ShcD mediates cell motility. 293 cell lines were altered to stably express GFP (GF) or GFP‑ShcD (G5). Treatment of the cells with transforming growth factor (TGF)β2 promoted extracellular signal‑regulated kinase (ERK) phosphorylation and, to a lesser extent, Smad2 phosphorylation in GFP‑ShcD‑expressing cells but not in GFP‑overexpressing cells. GFP‑ShcD‑expressing cells exhibited upregulated expression of certain epithelial‑mesenchymal transition‑related genes, such as snail family transcriptional repressor 1 and SLUG, than GFP‑expressing cells. Higher levels of ERK were found in the nuclear fraction of GFP‑ShcD‑expressing cells than that of GFP‑expressing cells. Overall, GFP‑ShcD‑expressing cells demonstrated enhanced migration compared with GFP‑expressing cells. A slight increase in cell migration was observed in both cell lines (GF and G5) when the cells were allowed to migrate towards conditioned medium derived from TGFβ2‑treated GFP‑ShcD expressing cells. Collectively, ShcD upregulation was proposed to induce cell migration by affecting the expression of certain epithelial‑mesenchymal transition‑related genes. Thus, our findings may improve understanding of the role of ShcD in cell migration.

References

1 

Takahashi Y, Tobe K, Kadowaki H, Katsumata D, Fukushima Y, Yazaki Y, Akanuma Y and Kadowaki T: Roles of insulin receptor substrate-1 and Shc on insulin-like growth factor I receptor signaling in early passages of cultured human fibroblasts. Endocrinology. 138:741–150. 1997. View Article : Google Scholar : PubMed/NCBI

2 

Polk DB: Shc is a substrate of the rat intestinal epidermal growth factor receptor tyrosine kinase. Gastroenterology. 109:1845–1851. 1995. View Article : Google Scholar : PubMed/NCBI

3 

Stephens RM, Loeb DM, Copeland TD, Pawson T, Greene LA and Kaplan DR: Trk receptors use redundant signal transduction pathways involving SHC and PLC-gamma 1 to mediate NGF responses. Neuron. 12:691–705. 1994. View Article : Google Scholar : PubMed/NCBI

4 

Ahmed SBM and Prigent SA: Insights into the Shc family of adaptor proteins. J Mol Signal. 12:22017. View Article : Google Scholar : PubMed/NCBI

5 

Ravichandran KS: Signaling via Shc family adapter proteins. Oncogene. 20:6322–6330. 2001. View Article : Google Scholar : PubMed/NCBI

6 

Fagiani E, Giardina G, Luzi L, Cesaroni M, Quarto M, Capra M, Germano G, Bono M, Capillo M, Pelicci P and Lanfrancone L: RaLP, a new member of the Src homology and collagen family, regulates cell migration and tumor growth of metastatic melanomas. Cancer Res. 67:3064–3073. 2007. View Article : Google Scholar : PubMed/NCBI

7 

Jones N, Hardy WR, Friese MB, Jorgensen C, Smith MJ, Woody NM, Burden SJ and Pawson T: Analysis of a Shc family adaptor protein, ShcD/Shc4, that associates with muscle-specific kinase. Mol Cell Biol. 27:4759–4773. 2007. View Article : Google Scholar : PubMed/NCBI

8 

Wells A, Chao YL, Grahovac J, Wu Q and Lauffenburger DA: Epithelial and mesenchymal phenotypic switchings modulate cell motility in metastasis. Front Biosci (Landmark Ed). 16:815–837. 2011. View Article : Google Scholar : PubMed/NCBI

9 

Kalluri R and Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 119:1420–1428. 2009. View Article : Google Scholar : PubMed/NCBI

10 

Son H and Moon A: Epithelial-mesenchymal transition and cell invasion. Toxicol Res. 26:245–252. 2010. View Article : Google Scholar : PubMed/NCBI

11 

Xie L, Law BK, Chytil AM, Brown KA, Aakre ME and Moses HL: Activation of the Erk pathway is required for TGF-beta1-induced EMT in vitro. Neoplasia. 6:603–610. 2004. View Article : Google Scholar : PubMed/NCBI

12 

Ferrari G, Cook BD, Terushkin V, Pintucci G and Mignatti P: Transforming growth factor-beta 1 (TGF-beta1) induces angiogenesis through vascular endothelial growth factor (VEGF)-mediated apoptosis. J Cell Physiol. 219:449–458. 2009. View Article : Google Scholar : PubMed/NCBI

13 

Xu J, Lamouille S and Derynck R: TGF-beta-induced epithelial to mesenchymal transition. Cell Res. 19:156–172. 2009. View Article : Google Scholar : PubMed/NCBI

14 

Zhang YE: Non-Smad pathways in TGF-beta signaling. Cell Res. 19:128–139. 2009. View Article : Google Scholar : PubMed/NCBI

15 

Hata A and Chen YG: TGF-β signaling from receptors to Smads. Cold Spring Harb Perspect Biol. 8(pii): a0220612016. View Article : Google Scholar : PubMed/NCBI

16 

Denis JF, Sader F, Gatien S, Villiard é, Philip A and Roy S: Activation of Smad2 but not Smad3 is required to mediate TGF-β signaling during axolotl limb regeneration. Development. 143:3481–3490. 2016. View Article : Google Scholar : PubMed/NCBI

17 

Naber HP, Drabsch Y, Snaar-Jagalska BE, ten Dijke P and van Laar T: Snail and Slug, key regulators of TGF-β-induced EMT, are sufficient for the induction of single-cell invasion. Biochem Biophys Res Commun. 435:58–63. 2013. View Article : Google Scholar : PubMed/NCBI

18 

Kim ES, Sohn YW and Moon A: TGF-beta-induced transcriptional activation of MMP-2 is mediated by activating transcription factor (ATF)2 in human breast epithelial cells. Cancer Lett. 252:147–156. 2007. View Article : Google Scholar : PubMed/NCBI

19 

Ahmed SBM, Amer S, Emad M, Rahmani M and Prigent SA: Studying the ShcD and ERK interaction under acute oxidative stress conditions in melanoma cells. Int J Biochem Cell Biol. 112:123–133. 2019. View Article : Google Scholar : PubMed/NCBI

20 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI

21 

Ahmed SB and Prigent SA: A nuclear export signal and oxidative stress regulate ShcD subcellular localisation: A potential role for ShcD in the nucleus. Cell Signal. 26:32–40. 2014. View Article : Google Scholar : PubMed/NCBI

22 

Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, et al: Fiji: An open-source platform for biological-image analysis. Nat Methods. 9:676–682. 2012. View Article : Google Scholar : PubMed/NCBI

23 

Friedl P and Bröcker EB: The biology of cell locomotion within three-dimensional extracellular matrix. Cell Mol Life Sci. 57:41–64. 2000. View Article : Google Scholar : PubMed/NCBI

24 

Dong W, Li H, Zhang Y, Yang H, Guo M, Li L and Liu T: Matrix metalloproteinase 2 promotes cell growth and invasion in colorectal cancer. Acta Biochim Biophys Sin (Shanghai). 43:840–848. 2011. View Article : Google Scholar : PubMed/NCBI

25 

Zetter BR: Angiogenesis and tumor metastasis. Annu Rev Med. 49:407–424. 1998. View Article : Google Scholar : PubMed/NCBI

26 

Fantozzi A, Gruber DC, Pisarsky L, Heck C, Kunita A, Yilmaz M, Meyer-Schaller N, Cornille K, Hopfer U, Bentires-Alj M and Christofori G: VEGF-mediated angiogenesis links EMT-induced cancer stemness to tumor initiation. Cancer Res. 74:1566–1575. 2014. View Article : Google Scholar : PubMed/NCBI

27 

Lee MK, Pardoux C, Hall MC, Lee PS, Warburton D, Qing J, Smith SM and Derynck R: TGF-beta activates Erk MAP kinase signalling through direct phosphorylation of ShcA. EMBO J. 26:3957–3967. 2007. View Article : Google Scholar : PubMed/NCBI

28 

Northey JJ, Chmielecki J, Ngan E, Russo C, Annis MG, Muller WJ and Siegel PM: Signaling through ShcA is required for transforming growth factor beta- and Neu/ErbB-2-induced breast cancer cell motility and invasion. Mol Cell Biol. 28:3162–3176. 2008. View Article : Google Scholar : PubMed/NCBI

29 

Hudson J, Ha JR, Sabourin V, Ahn R, La Selva R, Livingstone J, Podmore L, Knight J, Forrest L, Beauchemin N, et al: p66ShcA promotes breast cancer plasticity by inducing an epithelial-to-mesenchymal transition. Mol Cell Biol. 34:3689–3701. 2014. View Article : Google Scholar : PubMed/NCBI

30 

Navandar M, Garding A, Sahu SK, Pataskar A, Schick S and Tiwari VK: ERK signalling modulates epigenome to drive epithelial to mesenchymal transition. Oncotarget. 8:29269–29281. 2017. View Article : Google Scholar : PubMed/NCBI

31 

Chiu LY, Hsin IL, Yang TY, Sung WW, Chi JY, Chang JT, Ko JL and Sheu GT: The ERK-ZEB1 pathway mediates epithelial-mesenchymal transition in pemetrexed resistant lung cancer cells with suppression by vinca alkaloids. Oncogene. 36:242–253. 2017. View Article : Google Scholar : PubMed/NCBI

32 

Zheng H, Li W, Wang Y, Liu Z, Cai Y, Xie T, Shi M, Wang Z and Jiang B: Glycogen synthase kinase-3 beta regulates Snail and β-catenin expression during Fas-induced epithelial-mesenchymal transition in gastrointestinal cancer. Eur J Cancer. 49:2734–2746. 2013. View Article : Google Scholar : PubMed/NCBI

33 

Barrallo-Gimeno A and Nieto MA: The Snail genes as inducers of cell movement and survival: Implications in development and cancer. Development. 132:3151–3161. 2005. View Article : Google Scholar : PubMed/NCBI

34 

Ganesan R, Mallets E and Gomez-Cambronero J: The transcription factors Slug (SNAI2) and Snail (SNAI1) regulate phospholipase D (PLD) promoter in opposite ways towards cancer cell invasion. Mol Oncol. 10:663–676. 2016. View Article : Google Scholar : PubMed/NCBI

35 

Medici D, Hay ED and Olsen BR: Snail and Slug promote epithelial-mesenchymal transition through beta-catenin-T-cell factor-4-dependent expression of transforming growth factor-beta3. Mol Biol Cell. 19:4875–4887. 2008. View Article : Google Scholar : PubMed/NCBI

36 

Uygur B and Wu WS: SLUG promotes prostate cancer cell migration and invasion via CXCR4/CXCL12 axis. Mol Cancer. 10:1392011. View Article : Google Scholar : PubMed/NCBI

37 

Sun Y, Song GD, Sun N, Chen JQ and Yang SS: Slug overexpression induces stemness and promotes hepatocellular carcinoma cell invasion and metastasis. Oncol Lett. 7:1936–1940. 2014. View Article : Google Scholar : PubMed/NCBI

38 

Page-McCaw A, Ewald AJ and Werb Z: Matrix metalloproteinases and the regulation of tissue remodelling. Nat Rev Mol Cell Biol. 8:221–233. 2007. View Article : Google Scholar : PubMed/NCBI

39 

Lochter A, Galosy S, Muschler J, Freedman N, Werb Z and Bissell MJ: Matrix metalloproteinase stromelysin-1 triggers a cascade of molecular alterations that leads to stable epithelial-to-mesenchymal conversion and a premalignant phenotype in mammary epithelial cells. J Cell Biol. 139:1861–1872. 1997. View Article : Google Scholar : PubMed/NCBI

40 

Zhang D, Bar-Eli M, Meloche S and Brodt P: Dual regulation of MMP-2 expression by the type 1 insulin-like growth factor receptor: the phosphatidylinositol 3-kinase/Akt and Raf/ERK pathways transmit opposing signals. J Biol Chem. 279:19683–19690. 2004. View Article : Google Scholar : PubMed/NCBI

41 

Merikallio H, T TT, Pääkkö P, Mäkitaro R, Kaarteenaho R, Lehtonen S, Salo S, Salo T, Harju T and Soini Y: Slug is associated with poor survival in squamous cell carcinoma of the lung. Int J Clin Exp Pathol. 7:5846–5854. 2014.PubMed/NCBI

42 

Li Y, Klausen C, Zhu H and Leung PC: Activin a increases human trophoblast invasion by inducing SNAIL-mediated MMP2 Up-regulation through ALK4. J Clin Endocrinol Metab. 100:E1415–E1427. 2015. View Article : Google Scholar : PubMed/NCBI

43 

Jung YD, Nakano K, Liu W, Gallick GE and Ellis LM: Extracellular signal-regulated kinase activation is required for up-regulation of vascular endothelial growth factor by serum starvation in human colon carcinoma cells. Cancer Res. 59:4804–4807. 1999.PubMed/NCBI

Related Articles

Journal Cover

September 2019
Volume 20 Issue 3

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Amer, S., Alsayegh, F., Mashaal, Z., Mohamed, S., Shawa, N., Rajan, K., & Ahmed, S.B. (2019). Role of TGF‑β in the motility of ShcD‑overexpressing 293 cells. Molecular Medicine Reports, 20, 2667-2674. https://doi.org/10.3892/mmr.2019.10517
MLA
Amer, S., Alsayegh, F., Mashaal, Z., Mohamed, S., Shawa, N., Rajan, K., Ahmed, S. B."Role of TGF‑β in the motility of ShcD‑overexpressing 293 cells". Molecular Medicine Reports 20.3 (2019): 2667-2674.
Chicago
Amer, S., Alsayegh, F., Mashaal, Z., Mohamed, S., Shawa, N., Rajan, K., Ahmed, S. B."Role of TGF‑β in the motility of ShcD‑overexpressing 293 cells". Molecular Medicine Reports 20, no. 3 (2019): 2667-2674. https://doi.org/10.3892/mmr.2019.10517