Wnt3a increases the metastatic potential of non-small cell lung cancer cells in vitro in part via its upregulation of Notch3

Li,Chunyan; Song,Gongru; Zhang,Siyang; Wang,Enhua; Cui,Zeshi

doi:10.3892/or.2014.3700

Wnt3a increases the metastatic potential of non-small cell lung cancer cells in vitro in part via its upregulation of Notch3

Authors:
- Chunyan Li
- Gongru Song
- Siyang Zhang
- Enhua Wang
- Zeshi Cui
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Affiliations: Center of Laboratory Technology and Experimental Medicine, China Medical University, Shenyang, Liaoning 110001, P.R. China, Department of Biotechnology, China Medical University, Shenyang, Liaoning 110001, P.R. China, Department of Pathology, China Medical University, Shenyang, Liaoning 110001, P.R. China
Published online on: December 30, 2014 https://doi.org/10.3892/or.2014.3700
Pages: 1207-1214

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Abstract

Metastasis is the leading cause of death in lung cancer. Understanding the mechanisms underlying the process of metastasis is crucial for identifying novel anti-metastatic therapies. Studies indicate that the highly conserved developmental pathways, such as the Wnt and Notch signaling pathways, play important roles in the non-small cell lung cancer (NSCLC) tumorigenesis. However, the roles of both pathways in NSCLC metastasis are unclear. The present study aimed to investigate whether Wnt3a and Notch3, key components of the Wnt and Notch signaling pathways, respectively, regulate the metastatic abilities of NSCLC cells and whether there is some relationship during these regulatory events. Here, we observed that Wnt3a treatment upregulated, not only the protein expression of Notch3, but also the mRNA expression of Notch3 and its downstream genes, HES1 and HEYL. In addition, Wnt3a promoted cell invasion and anchorage-independent growth. Meanwhile, Wnt3a treatment caused epithelial‑mesenchymal transition (EMT)-like morphological changes and F-actin reorganization. The western blotting data showed that Wnt3a treatment decreased the expression of E-cadherin and increased the expression of N-cadherin and vimentin. Compared with Wnt3a treatment, Notch3 shRNA transfection had opposite effects. Furthermore, Notch3 shRNA weakened the effects of Wnt3a treatment on the in vitro cell invasion and EMT. Overall, these observations suggest that Wnt3a and Notch3 may promote the metastasis of NSCLC and Notch3 upregulation is required for the Wnt3a mediated increased metastatic abilities of NSCLC.

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1	Perlikos F, Harrington KJ and Syrigos KN: Key molecular mechanisms in lung cancer invasion and metastasis: A comprehensive review. Crit Rev Oncol Hematol. 87:1–11. 2013. View Article : Google Scholar : PubMed/NCBI
2	Xie C, Jiang G, Fan C, et al: ARMC8α promotes proliferation and invasion of non-small cell lung cancer cells by activating the canonical Wnt signaling pathway. Tumour Biol. 35:8903–8911. 2014. View Article : Google Scholar : PubMed/NCBI
3	Gao Y, Song C, Hui L, et al: Overexpression of RNF146 in non-small cell lung cancer enhances proliferation and invasion of tumors through the Wnt/β-catenin signaling pathway. PLoS One. 9:e853772014. View Article : Google Scholar
4	Su K, Huang L, Li W, et al: TC-1 (c8orf4) enhances aggressive biologic behavior in lung cancer through the Wnt/β-catenin pathway. J Surg Res. 185:255–263. 2013. View Article : Google Scholar : PubMed/NCBI
5	Zhang S, Wang Y, Dai SD and Wang EH: Down-regulation of NKD1 increases the invasive potential of non-small-cell lung cancer and correlates with a poor prognosis. BMC Cancer. 11:1862011. View Article : Google Scholar : PubMed/NCBI
6	Ren J, Wang R, Huang G, Song H, Chen Y and Chen L: sFRP1 inhibits epithelial-mesenchymal transition in A549 human lung adenocarcinoma cell line. Cancer Biother Radiopharm. 28:565–571. 2013. View Article : Google Scholar : PubMed/NCBI
7	Singh T and Katiyar SK: Honokiol inhibits non-small cell lung cancer cell migration by targeting PGE₂-mediated activation of β-catenin signaling. PLoS One. 8:e607492013. View Article : Google Scholar
8	Westhoff B, Colaluca IN, D’Ario G, et al: Alterations of the Notch pathway in lung cancer. Proc Natl Acad Sci USA. 106:22293–22298. 2009. View Article : Google Scholar : PubMed/NCBI
9	Liu L, Chen X, Wang Y, et al: Notch3 is important for TGF-β-induced epithelial-mesenchymal transition in non-small cell lung cancer bone metastasis by regulating ZEB-1. Cancer Gene Ther. 21:364–372. 2014. View Article : Google Scholar : PubMed/NCBI
10	Li C, Zhang S, Lu Y, Zhang Y, Wang E and Cui Z: The roles of Notch3 on the cell proliferation and apoptosis induced by CHIR99021 in NSCLC cell lines: a functional link between Wnt and Notch signaling pathways. PLoS One. 8:e846592013. View Article : Google Scholar : PubMed/NCBI
11	Gao CF, Xie Q, Su YL, et al: Proliferation and invasion: plasticity in tumor cells. Proc Natl Acad Sci USA. 102:10528–10533. 2005. View Article : Google Scholar : PubMed/NCBI
12	Al-Mehdi AB, Tozawa K, Fisher AB, Shientag L, Lee A and Muschel RJ: Intravascular origin of metastasis from the proliferation of endothelium-attached tumor cells: a new model for metastasis. Nat Med. 6:100–102. 2000. View Article : Google Scholar
13	Kikuchi A, Yamamoto H and Kishida S: Multiplicity of the interactions of Wnt proteins and their receptors. Cell Signal. 19:659–671. 2007. View Article : Google Scholar
14	Stapp AD, Gómez BI, Gifford CA, Hallford DM and Hernandez Gifford JA: Canonical WNT signaling inhibits follicle stimulating hormone mediated steroidogenesis in primary cultures of rat granulosa cells. PLoS One. 9:e864322014. View Article : Google Scholar : PubMed/NCBI
15	Yi F, Amarasinghe B and Dang TP: Manic fringe inhibits tumor growth by suppressing Notch3 degradation in lung cancer. Am J Cancer Res. 3:490–499. 2013.PubMed/NCBI
16	Zheng Y, de la Cruz CC, Sayles LC, et al: A rare population of CD24(+)ITGB4(+)Notch(hi) cells drives tumor propagation in NSCLC and requires Notch3 for self-renewal. Cancer Cell. 24:59–74. 2013. View Article : Google Scholar : PubMed/NCBI
17	Frisch SM and Francis H: Disruption of epithelial cell-matrix interactions induces apoptosis. J Cell Biol1. 24:619–626. 1994. View Article : Google Scholar
18	Nomura Y, Tashiro H and Hisamatsu K: In vitro clonogenic growth and metastatic potential of human operable breast cancer. Cancer Res. 49:5288–5293. 1989.PubMed/NCBI
19	Nakanishi K, Sakamoto M, Yasuda J, et al: Critical involvement of the phosphatidylinositol 3-kinase/Akt pathway in anchorage-independent growth and hematogeneous intrahepatic metastasis of liver cancer. Cancer Res. 62:2971–2975. 2002.PubMed/NCBI
20	Tsai JH and Yang J: Epithelial-mesenchymal plasticity in carcinoma metastasis. Genes Dev. 27:2192–2206. 2013. View Article : Google Scholar : PubMed/NCBI
21	Huber MA, Kraut N and Beug H: Molecular requirements for epithelial-mesenchymal transition during tumor progression. Curr Opin Cell Biol. 17:548–558. 2005. View Article : Google Scholar : PubMed/NCBI
22	Gupta GP and Massagué J: Cancer metastasis: building a framework. Cell. 127:679–695. 2006. View Article : Google Scholar : PubMed/NCBI
23	Poste G and Fidler IJ: The pathogenesis of cancer metastasis. Nature. 283:139–146. 1980. View Article : Google Scholar : PubMed/NCBI
24	Bo H, Zhang S, Gao L, Chen Y, Zhang J, Chang X and Zhu M: Upregulation of Wnt5a promotes epithelial-to-mesenchymal transition and metastasis of pancreatic cancer cells. BMC Cancer. 13:4962013. View Article : Google Scholar : PubMed/NCBI
25	Ford CE, Jary E, Ma SS, Nixdorf S, Heinzelmann-Schwarz VA and Ward RL: The Wnt gatekeeper SFRP4 modulates EMT, cell migration and downstream Wnt signalling in serous ovarian cancer cells. PLoS One. 8:e543622013. View Article : Google Scholar : PubMed/NCBI
26	Kwon M, Lee SJ, Wang Y, et al: Filamin A interacting protein 1-like inhibits WNT signaling and MMP expression to suppress cancer cell invasion and metastasis. Int J Cancer. 135:48–60. 2014. View Article : Google Scholar
27	Jiang W, Crossman DK, Mitchell EH, Sohn P, Crowley MR and Serra R: WNT5A inhibits metastasis and alters splicing of Cd44 in breast cancer cells. PLoS One. 8:e583292013. View Article : Google Scholar : PubMed/NCBI
28	Dey N, Barwick BG, Moreno CS, et al: Wnt signaling in triple negative breast cancer is associated with metastasis. BMC Cancer. 13:5372013. View Article : Google Scholar : PubMed/NCBI
29	Espinoza I and Miele L: Deadly crosstalk: Notch signaling at the intersection of EMT and cancer stem cells. Cancer Lett. 341:41–45. 2013. View Article : Google Scholar : PubMed/NCBI
30	Espinoza I, Pochampally R, Xing F, Watabe K and Miele L: Notch signaling: targeting cancer stem cells and epithelial-to-mesenchymal transition. Onco Targets Ther. 6:1249–1259. 2013.PubMed/NCBI
31	Du R, Sun W, Xia L, et al: Hypoxia-induced down-regulation of microRNA-34a promotes EMT by targeting the Notch signaling pathway in tubular epithelial cells. PLoS One. 7:e307712012. View Article : Google Scholar : PubMed/NCBI
32	Zhang Q, Bai X, Chen W, et al: Wnt/β-catenin signaling enhances hypoxia-induced epithelial-mesenchymal transition in hepatocellular carcinoma via crosstalk with hif-1α signaling. Carcinogenesis. 34:962–973. 2013. View Article : Google Scholar : PubMed/NCBI
33	Zhai Y, Iura A, Yeasmin S, et al: MSX2 is an oncogenic downstream target of activated WNT signaling in ovarian endometrioid adenocarcinoma. Oncogene. 30:4152–4162. 2011. View Article : Google Scholar : PubMed/NCBI
34	Gupta N, Xu Z, El-Sehemy A, Steed H and Fu Y: Notch3 induces epithelial-mesenchymal transition and attenuates carboplatin-induced apoptosis in ovarian cancer cells. Gynecol Oncol. 130:200–206. 2013. View Article : Google Scholar : PubMed/NCBI
35	Katoh M and Katoh M: NUMB is a break of WNT-Notch signaling cycle. Int J Mol Med. 18:517–521. 2006.PubMed/NCBI
36	Bertrand FE, Angus CW, Partis WJ and Sigounas G: Developmental pathways in colon cancer: crosstalk between WNT, BMP, Hedgehog and Notch. Cell Cycle. 11:4344–4351. 2012. View Article : Google Scholar : PubMed/NCBI