TNF‑α inhibits xenograft tumor formation by A549 lung cancer cells in nude mice via the HIF‑1α/VASP signaling pathway

Liu,Wen; Chen,Xiang; He,Yanqi; Tian,Yihao; Xu,Liu; Ma,Yanbin; Hu,Pengchao; Su,Ke; Luo,Zhenyu; Wei,Lei; Zhang,Jingwei

doi:10.3892/or.2019.7026

TNF‑α inhibits xenograft tumor formation by A549 lung cancer cells in nude mice via the HIF‑1α/VASP signaling pathway

Authors:
- Wen Liu
- Xiang Chen
- Yanqi He
- Yihao Tian
- Liu Xu
- Yanbin Ma
- Pengchao Hu
- Ke Su
- Zhenyu Luo
- Lei Wei
- Jingwei Zhang
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Affiliations: Hubei Provincial Key Laboratory of Developmentally Originated Disease, Department of Pathology and Pathophysiology, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei 430071, P.R. China, Department of Human Anatomy and Histology and Embryology, School of Basic Medical Sciences, Wuhan University, Wuhan, Hubei 430071, P.R. China, Renmin Hospital of Wuhan University, The First College of Clinical Medicine of Wuhan University, Wuhan, Hubei 430060, P.R. China, Department of Breast and Thyroid Surgery, Zhongnan Hospital of Wuhan University, Hubei Key Laboratory of Tumor Biological Behaviors, Hubei Cancer Clinical Study Center, Wuhan, Hubei 430071, P.R. China
Published online on: February 21, 2019 https://doi.org/10.3892/or.2019.7026
Pages: 2418-2430

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Abstract

Lung cancer is the leading cause of cancer‑associated mortality worldwide. Tumor necrosis factor α (TNF‑α) is an important cytokine in the tumor microenvironment that serves a function in the balance of cell survival and cell death pathways. Our previous studies indicated that hypoxia‑inducible factor 1α (HIF‑1α) acts downstream of TNF‑α in MCF‑7 luminal breast cancer cells. However, whether vasodilator‑stimulated phosphoprotein (VASP) is implicated in the direct regulation of HIF‑1α in response to TNF‑α in lung cancer remains unknown. In vitro studies were performed using A549 and H226 lung carcinoma cells and in vivo studies of tumor xenograft models were performed to investigate the effects of TNF‑α. The results demonstrated that TNF‑α decreased VASP expression by upregulating the expression of HIF‑1α to inhibit A549 cell proliferation and adhesion. Inhibition of transplanted tumor growth was associated with downregulation of VASP expression in nude mice. Bioinformatics analysis indicated that expression levels of VASP or HIF‑1α lead to differential outcomes of overall survival in lung carcinoma. These results suggest that the HIF‑1α/VASP signaling pathway serves an important function in the regulation of TNF‑α‑induced suppression of A549 cell proliferation and xenograft growth. This may improve our understanding of the antitumor effect of TNF‑α.

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1	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
2	Zagryazhskaya A, Gyuraszova K and Zhivotovsky B: Cell death in cancer therapy of lung adenocarcinoma. Int J Dev Biol. 59:119–129. 2015. View Article : Google Scholar : PubMed/NCBI
3	Yatabe Y, Borczuk AC and Powell CA: Do all lung adenocarcinomas follow a stepwise progression? Lung Cancer. 74:7–11. 2011. View Article : Google Scholar : PubMed/NCBI
4	Chen ZY, Zhong WZ, Zhang XC, Su J, Yang XN, Chen ZH, Yang JJ, Zhou Q, Yan HH, An SJ, et al: EGFR mutation heterogeneity and the mixed response to EGFR tyrosine kinase inhibitors of lung adenocarcinomas. Oncologist. 17:978–985. 2012. View Article : Google Scholar : PubMed/NCBI
5	Devarakonda S, Morgensztern D and Govindan R: Genomic alterations in lung adenocarcinoma. Lancet Oncol. 16:e342–e351. 2015. View Article : Google Scholar : PubMed/NCBI
6	Ben-Baruch A: The tumor-promoting flow of cells into, within and out of the tumor site: Regulation by the inflammatory axis of TNF^α and chemokines. Cancer Microenviron. 5:151–164. 2012. View Article : Google Scholar : PubMed/NCBI
7	Wajant H: The role of TNF in cancer. Results Probl Cell Differ. 49:1–15. 2009. View Article : Google Scholar : PubMed/NCBI
8	Zelová H and Hošek J: TNF-^α signalling and inflammation: Interactions between old acquaintances. Inflamm Res. 62:641–651. 2013. View Article : Google Scholar : PubMed/NCBI
9	Wu Y and Zhou BP: TNF-alpha/NF-kappaB/Snail pathway in cancer cell migration and invasion. Br J Cancer. 102:639–644. 2010. View Article : Google Scholar : PubMed/NCBI
10	Carswell EA, Old LJ, Kassel RL, Green S, Fiore N and Williamson B: An endotoxin-induced serum factor that causes necrosis of tumors. Proc Natl Acad Sci USA. 72:3666–3670. 1975. View Article : Google Scholar : PubMed/NCBI
11	Haranaka K, Satomi N and Sakurai A: Antitumor activity of tumor necrosis factor (TNF) in vitro and in vivo. Gan To Kagaku Ryoho. 11:1387–1393. 1984.(In Japanese). PubMed/NCBI
12	Calzascia T, Pellegrini M, Hall H, Sabbagh L, Ono N, Elford AR, Mak TW and Ohashi PS: TNF-alpha is critical for antitumor but not antiviral T cell immunity in mice. J Clin Invest. 117:3833–3845. 2007.PubMed/NCBI
13	Fajardo LF, Kwan HH, Kowalski J, Prionas SD and Allison AC: Dual role of tumor necrosis factor-alpha in angiogenesis. Am J Pathol. 140:539–544. 1992.PubMed/NCBI
14	Weishaupt A, Gold R, Hartung T, Gaupp S, Wendel A, Brück W and Toyka KV: Role of TNF-alpha in high-dose antigen therapy in experimental autoimmune neuritis: Inhibition of TNF-alpha by neutralizing antibodies reduces T-cell apoptosis and prevents liver necrosis. J Neuropathol Exp Neurol. 59:368–376. 2000. View Article : Google Scholar : PubMed/NCBI
15	Bertazza L and Mocellin S: The dual role of tumor necrosis factor (TNF) in cancer biology. Curr Med Chem. 17:3337–3352. 2010. View Article : Google Scholar : PubMed/NCBI
16	Semenza GL and Wang GL: A nuclear factor induced by hypoxia via de novo protein synthesis binds to the human erythropoietin gene enhancer at a site required for transcriptional activation. Mol Cell Biol. 12:5447–5454. 1992. View Article : Google Scholar : PubMed/NCBI
17	Wang GL, Jiang BH, Rue EA and Semenza GL: Hypoxia-inducible factor 1 is a basic-helix-loop-helix-PAS heterodimer regulated by cellular O2 tension. Proc Natl Acad Sci USA. 92:5510–5514. 1995. View Article : Google Scholar : PubMed/NCBI
18	Lee JW, Bae SH, Jeong JW, Kim SH and Kim KW: Hypoxia-inducible factor (HIF-1)alpha: Its protein stability and biological functions. Exp Mol Med. 36:1–12. 2004. View Article : Google Scholar : PubMed/NCBI
19	Weidemann A and Johnson RS: Biology of HIF-1alpha. Cell Death Differ. 15:621–627. 2008. View Article : Google Scholar : PubMed/NCBI
20	Semenza GL: Targeting HIF-1 for cancer therapy. Nat Rev Cancer. 3:721–732. 2003. View Article : Google Scholar : PubMed/NCBI
21	Schindl M, Schoppmann SF, Samonigg H, Hausmaninger H, Kwasny W, Gnant M, Jakesz R, Kubista E, Birner P and Oberhuber G; Austrian Breast and Colorectal Cancer Study Group, : Overexpression of hypoxia-inducible factor 1alpha is associated with an unfavorable prognosis in lymph node-positive breast cancer. Clin Cancer Res. 8:1831–1837. 2002.PubMed/NCBI
22	Chiavarina B, Whitaker-Menezes D, Migneco G, Martinez-Outschoorn UE, Pavlides S, Howell A, Tanowitz HB, Casimiro MC, Wang C, Pestell RG, et al: HIF1-alpha functions as a tumor promoter in cancer associated fibroblasts, and as a tumor suppressor in breast cancer cells: Autophagy drives compartment-specific oncogenesis. Cell Cycle. 9:3534–3551. 2010. View Article : Google Scholar : PubMed/NCBI
23	Fan LF, Diao LM, Chen DJ, Liu MQ, Zhu LQ, Li HG, Tang ZJ, Xia D, Liu X and Chen HL: Expression of HIF-1 alpha and its relationship to apoptosis and proliferation in lung cancer. Ai Zheng. 21:254–258. 2002.(In Chinese). PubMed/NCBI
24	Balamurugan K: HIF-1 at the crossroads of hypoxia, inflammation, and cancer. Int J Cancer. 138:1058–1066. 2016. View Article : Google Scholar : PubMed/NCBI
25	Haffner C, Jarchau T, Reinhard M, Hoppe J, Lohmann SM and Walter U: Molecular cloning, structural analysis and functional expression of the proline-rich focal adhesion and microfilament-associated protein VASP. EMBO J. 14:19–27. 1995. View Article : Google Scholar : PubMed/NCBI
26	Pula G and Krause M: Role of Ena/VASP proteins in homeostasis and disease. Handb Exp Pharmacol. 39–65. 2008. View Article : Google Scholar : PubMed/NCBI
27	Chen XJ, Squarr AJ, Stephan R, Chen B, Higgins TE, Barry DJ, Martin MC, Rosen MK, Bogdan S and Way M: Ena/VASP proteins cooperate with the WAVE complex to regulate the actin cytoskeleton. Dev Cell. 30:569–584. 2014. View Article : Google Scholar : PubMed/NCBI
28	Zhang Y, Han G, Fan B, Zhou Y, Zhou X, Wei L and Zhang J: Green tea (−)-epigallocatechin-3-gallate down-regulates VASP expression and inhibits breast cancer cell migration and invasion by attenuating Rac1 activity. Eur J Pharmacol. 606:172–179. 2009. View Article : Google Scholar : PubMed/NCBI
29	Galler AB, García Arguinzonis MI, Baumgartner W, Kuhn M, Smolenski A, Simm A and Reinhard M: VASP-dependent regulation of actin cytoskeleton rigidity, cell adhesion, and detachment. Histochem Cell Biol. 125:457–474. 2006. View Article : Google Scholar : PubMed/NCBI
30	Knauer O, Binai NA, Carra G, Beckhaus T, Hanschmann KM, Renné T, Backert S, Karas M and Wessler S: Differential phosphoproteome profiling reveals a functional role for VASP in Helicobacter pylori-induced cytoskeleton turnover in gastric epithelial cells. Cell Microbiol. 10:2285–2296. 2008. View Article : Google Scholar : PubMed/NCBI
31	Liu K, Li L, Nisson PE, Gruber C, Jessee J and Cohen SN: Reversible tumorigenesis induced by deficiency of vasodilator-stimulated phosphoprotein. Mol Cell Biol. 19:3696–3703. 1999. View Article : Google Scholar : PubMed/NCBI
32	Su K, Hu P, Wang X, Kuang C, Xiang Q, Yang F, Xiang J, Zhu S, Wei L and Zhang J: Tumor suppressor berberine binds VASP to inhibit cell migration in basal-like breast cancer. Oncotarget. 7:45849–45862. 2016. View Article : Google Scholar : PubMed/NCBI
33	Su K, Tian Y, Wang J, Shi W, Luo D, Liu J, Tong Z, Wu J, Zhang J and Wei L: HIF-1^α acts downstream of TNF-^α to inhibit vasodilator-stimulated phosphoprotein expression and modulates the adhesion and proliferation of breast cancer cells. DNA Cell Biol. 31:1078–1087. 2012. View Article : Google Scholar : PubMed/NCBI
34	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
35	Wang Y, Hu PC, Gao FF, Lv JW, Xu S, Kuang CC, Wei L and Zhang JW: The protective effect of curcumin on hepatotoxicity and ultrastructural damage induced by cisplatin. Ultrastruct Pathol. 38:358–362. 2014. View Article : Google Scholar : PubMed/NCBI
36	Dertsiz L, Ozbilim G, Kayisli Y, Gokhan GA, Demircan A and Kayisli UA: Differential expression of VASP in normal lung tissue and lung adenocarcinomas. Thorax. 60:576–581. 2005. View Article : Google Scholar : PubMed/NCBI
37	Gyorffy B, Surowiak P, Budczies J and Lanczky A: Online survival analysis software to assess the prognostic value of biomarkers using transcriptomic data in non-small-cell lung cancer. PLoS One. 8:e822412013. View Article : Google Scholar : PubMed/NCBI
38	Wang X and Adjei AA: Lung cancer and metastasis: New opportunities and challenges. Cancer Metastasis Rev. 34:169–171. 2015. View Article : Google Scholar : PubMed/NCBI
39	Baxevanis CN, Voutsas IF, Tsitsilonis OE, Tsiatas ML, Gritzapis AD and Papamichail M: Compromised anti-tumor responses in tumor necrosis factor-alpha knockout mice. Eur J Immunol. 30:1957–1966. 2000. View Article : Google Scholar : PubMed/NCBI
40	Prevost-Blondel A, Roth E, Rosenthal FM and Pircher H: Crucial role of TNF-alpha in CD8 T cell-mediated elimination of 3LL-A9 Lewis lung carcinoma cells in vivo. J Immunol. 164:3645–3651. 2000. View Article : Google Scholar : PubMed/NCBI
41	Tandle A, Hanna E, Lorang D, Hajitou A, Moya CA, Pasqualini R, Arap W, Adem A, Starker E, Hewitt S and Libutti SK: Tumor vasculature-targeted delivery of tumor necrosis factor-alpha. Cancer. 115:128–139. 2009. View Article : Google Scholar : PubMed/NCBI
42	Wang J, Zhang J, Wu J, Luo D, Su K, Shi W, Liu J, Tian Y and Wei L: MicroRNA-610 inhibits the migration and invasion of gastric cancer cells by suppressing the expression of vasodilator-stimulated phosphoprotein. Eur J Cancer. 48:1904–1913. 2012. View Article : Google Scholar : PubMed/NCBI
43	Henes J, Schmit MA, Morote-Garcia JC, Mirakaj V, Köhler D, Glover L, Eldh T, Walter U, Karhausen J, Colgan SP and Rosenberger P: Inflammation-associated repression of vasodilator-stimulated phosphoprotein (VASP) reduces alveolar-capillary barrier function during acute lung injury. FASEB J. 23:4244–4255. 2009. View Article : Google Scholar : PubMed/NCBI
44	Azoitei N, Becher A, Steinestel K, Rouhi A, Diepold K, Genze F, Simmet T and Seufferlein T: PKM2 promotes tumor angiogenesis by regulating HIF-1^α through NF-κB activation. Mol Cancer. 15:32016. View Article : Google Scholar : PubMed/NCBI
45	Carmeliet P, Dor Y, Herbert JM, Fukumura D, Brusselmans K, Dewerchin M, Neeman M, Bono F, Abramovitch R, Maxwell P, et al: Role of HIF-1alpha in hypoxia-mediated apoptosis, cell proliferation and tumour angiogenesis. Nature. 394:485–490. 1998. View Article : Google Scholar : PubMed/NCBI
46	Haddad JJ and Harb HL: Cytokines and the regulation of hypoxia-inducible factor (HIF)-1alpha. Int Immunopharmacol. 5:461–483. 2005. View Article : Google Scholar : PubMed/NCBI
47	Zhang H, Wong CC, Wei H, Gilkes DM, Korangath P, Chaturvedi P, Schito L, Chen J, Krishnamachary B, Winnard PT Jr, et al: HIF-1-dependent expression of angiopoietin-like 4 and L1CAM mediates vascular metastasis of hypoxic breast cancer cells to the lungs. Oncogene. 31:1757–1770. 2012. View Article : Google Scholar : PubMed/NCBI