EHF enhances malignancy by modulating AKT and MAPK/ERK signaling in non‑small cell lung cancer cells

Gao,Lei; Yang,Tian; Zhang,Shuo; Liang,Yiqian; Shi,Puyu; Ren,Hui; Hou,Peng; Chen,Mingwei

doi:10.3892/or.2021.8053

EHF enhances malignancy by modulating AKT and MAPK/ERK signaling in non‑small cell lung cancer cells

Authors:
- Lei Gao
- Tian Yang
- Shuo Zhang
- Yiqian Liang
- Puyu Shi
- Hui Ren
- Peng Hou
- Mingwei Chen
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Affiliations: Department of Endocrinology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China, Shanxi Provincial Research Center for The Project of Prevention and Treatment of Respiratory Diseases, Xi'an, Shaanxi 710061, P.R. China, Department of Endocrinology, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China
Published online on: April 15, 2021 https://doi.org/10.3892/or.2021.8053
Article Number: 102
Copyright: © Gao et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Overexpression of ETS‑homologous factor (EHF) in non‑small cell lung cancer (NSCLC) is associated with poor patient prognosis. To explore the mechanism of the effect of EHF in NSCLC, EHF expression was examined in NSCLC and its role in cell proliferation, invasion, cell cycle, and apoptosis of NSCLC cells was evaluated by overexpressing EHF and/or knocking down EHF expression in NSCLC cells in vitro and in cancer cell grafted mice in vivo. The results revealed that the knockdown of EHF expression in NSCLC with siRNA significantly inhibited cell proliferation and invasion, arrested the cell cycle at the G₀/G₁ phase, and induced apoptosis, whereas overexpression of EHF in NSCLC promoted cell proliferation, tumor growth, and cancer cell migration in vitro. The in vivo experiments demonstrated that siRNA‑mediated downregulation of EHF expression in NSCLC cells significantly suppressed tumor growth in xenografted nude mice as compared to cancer progression in the mice grafted with NSCLC cells transfected with non‑specific control siRNA. The biochemical analyses revealed that EHF promoted NSCLC growth by regulating the transcription of Erb‑B2 receptor tyrosine kinase 2/3 (ERBB2, ERBB3) and mesenchymal‑epithelial transition (MET) factor tyrosine kinase receptors and modulating the AKT and ERK signaling pathways in the NSCLC cells. The present findings indicated that EHF could be used as a prognostic marker for NSCLC, and tyrosine kinase receptors of ERBB2, ERBB3 and MET could be drug targets for NSCLC treatment.

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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	Devesa SS, Bray F, Vizcaino AP and Parkin DM: International lung cancer trends by histologic type: Male:Female differences diminishing and adenocarcinoma rates rising. Int J Cancer. 117:294–299. 2005. View Article : Google Scholar : PubMed/NCBI
3	Jemal A, Bray F, Center MM, Ferlay J, Ward E and Forman D: Global cancer statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar : PubMed/NCBI
4	Sementchenko VI and Watson DK: Ets target genes: Past, present and future. Oncogene. 19:6533–6548. 2000. View Article : Google Scholar : PubMed/NCBI
5	Seth A and Watson DK: ETS transcription factors and their emerging roles in human cancer. Eur J Cancer. 41:2462–2478. 2005. View Article : Google Scholar : PubMed/NCBI
6	Wasylyk B, Hagman J and Gutierrez-Hartmann A: Ets transcription factors: Nuclear effectors of the Ras-MAP-kinase signaling pathway. Trends Biochem Sci. 23:213–216. 1998. View Article : Google Scholar : PubMed/NCBI
7	Trojanowska M: Ets factors and regulation of the extracellular matrix. Oncogene. 19:6464–6471. 2000. View Article : Google Scholar : PubMed/NCBI
8	Benz CC, O'Hagan RC, Richter B, Scott GK, Chang CH, Xiong X, Chew K, Ljung BM, Edgerton S, Thor A and Hassell JA: HER2/Neu and the Ets transcription activator PEA3 are coordinately upregulated in human breast cancer. Oncogene. 15:1513–1525. 1997. View Article : Google Scholar : PubMed/NCBI
9	Eckel KL, Tentler JJ, Cappetta GJ, Diamond SE and Gutierrez-Hartmann A: The epithelial-specific ETS transcription factor ESX/ESE-1/Elf-3 modulates breast cancer-associated gene expression. DNA Cell Biol. 22:79–94. 2003. View Article : Google Scholar : PubMed/NCBI
10	Kas K, Finger E, Grall F, Gu X, Akbarali Y, Boltax J, Weiss A, Oettgen P, Kapeller R and Libermann TA: ESE-3, a novel member of an epithelium-specific ets transcription factor subfamily, demonstrates different target gene specificity from ESE-1. J Biol Chem. 275:2986–2998. 2000. View Article : Google Scholar : PubMed/NCBI
11	Kleinbaum LA, Duggan C, Ferreira E, Coffey GP, Buttice G and Burton FH: Human chromosomal localization, tissue/tumor expression, and regulatory function of the ets family gene EHF. Biochem Biophys Res Commun. 264:119–126. 1999. View Article : Google Scholar : PubMed/NCBI
12	Cangemi R, Mensah A, Albertini V, Jain A, Mello-Grand M, Chiorino G, Catapano CV and Carbone GM: Reduced expression and tumor suppressor function of the ETS transcription factor ESE-3 in prostate cancer. Oncogene. 27:2877–2885. 2008. View Article : Google Scholar : PubMed/NCBI
13	Shaikhibrahim Z, Lindstrot A, Langer B, Buettner R and Wernert N: Differential expression of ETS family members in prostate cancer tissues and androgen-sensitive and insensitive prostate cancer cell lines. Int J Mol Med. 28:89–93. 2011.PubMed/NCBI
14	Turcotte S, Forget MA, Beauseigle D, Nassif E and Lapointe R: Prostate-derived Ets transcription factor overexpression is associated with nodal metastasis and hormone receptor positivity in invasive breast cancer. Neoplasia. 9:788–796. 2007. View Article : Google Scholar : PubMed/NCBI
15	Brenne K, Nymoen DA, Hetland TE, Trope CG and Davidson B: Expression of the Ets transcription factor EHF in serous ovarian carcinoma effusions is a marker of poor survival. Hum Pathol. 43:496–505. 2012. View Article : Google Scholar : PubMed/NCBI
16	Hynes NE and MacDonald G: ErbB receptors and signaling pathways in cancer. Curr Opin Cell Biol. 21:177–184. 2009. View Article : Google Scholar : PubMed/NCBI
17	Gelsomino F, Facchinetti F, Haspinger ER, Garassino MC, Trusolino L, De Braud F and Tiseo M: Targeting the MET gene for the treatment of non-small-cell lung cancer. Crit Rev Oncol Hematol. 89:284–299. 2014. View Article : Google Scholar : PubMed/NCBI
18	Zhang S, Gao L, Thakur A, Shi P, Liu F, Feng J, Wang T, Liang Y, Liu JJ, Chen M and Ren H: MiRNA-204 suppresses human non-small cell lung cancer by targeting ATF2. Tumour Biol. 37:11177–11186. 2016. View Article : Google Scholar : PubMed/NCBI
19	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
20	Wang P, Henning SM and Heber D: Limitations of MTT and MTS-based assays for measurement of antiproliferative activity of green tea polyphenols. PLoS One. 5:e102022010. View Article : Google Scholar : PubMed/NCBI
21	Shi J, Liu W, Sui F, Lu R, He Q, Yang Q, Lv H, Shi B and Hou P: Frequent amplification of AIB1, a critical oncogene modulating major signaling pathways, is associated with poor survival in gastric cancer. Oncotarget. 6:14344–14359. 2015. View Article : Google Scholar : PubMed/NCBI
22	Hollenhorst PC, Jones DA and Graves BJ: Expression profiles frame the promoter specificity dilemma of the ETS family of transcription factors. Nucleic Acids Res. 32:5693–5702. 2004. View Article : Google Scholar : PubMed/NCBI
23	Roskoski R Jr: The ErbB/HER family of protein-tyrosine kinases and cancer. Pharmacol Res. 79:34–74. 2014. View Article : Google Scholar : PubMed/NCBI
24	Holbro T, Civenni G and Hynes NE: The ErbB receptors and their role in cancer progression. Exp Cell Res. 284:99–110. 2003. View Article : Google Scholar : PubMed/NCBI
25	Roskoski R Jr: The ErbB/HER receptor protein-tyrosine kinases and cancer. Biochem Biophys Res Commun. 319:1–11. 2004. View Article : Google Scholar : PubMed/NCBI
26	Petrini I: Biology of MET: A double life between normal tissue repair and tumor progression. Ann Transl Med. 3:822015.PubMed/NCBI
27	Cappuzzo F, Marchetti A, Skokan M, Rossi E, Gajapathy S, Felicioni L, Del Grammastro M, Sciarrotta MG, Buttitta F, Incarbone M, et al: Increased MET gene copy number negatively affects survival of surgically resected non-small-cell lung cancer patients. J Clin Oncol. 27:1667–1674. 2009. View Article : Google Scholar : PubMed/NCBI
28	Fu P, Du F, Yao M, Lv K and Liu Y: MicroRNA-185 inhibits proliferation by targeting c-Met in human breast cancer cells. Exp Ther Med. 8:1879–1883. 2014. View Article : Google Scholar : PubMed/NCBI
29	Hagman Z, Haflidadottir BS, Ansari M, Persson M, Bjartell A, Edsjö A and Ceder Y: The tumour suppressor miR-34c targets MET in prostate cancer cells. Br J Cancer. 109:1271–1278. 2013. View Article : Google Scholar : PubMed/NCBI
30	Phan LM, Fuentes-Mattei E, Wu W, Velazquez-Torres G, Sircar K, Wood CG, Hai T, Jimenez C, Cote GJ, Ozsari L, et al: Hepatocyte Growth Factor/cMET pathway activation enhances cancer hallmarks in adrenocortical carcinoma. Cancer Res. 75:4131–4142. 2015. View Article : Google Scholar : PubMed/NCBI
31	Maroun CR and Rowlands T: The Met receptor tyrosine kinase: A key player in oncogenesis and drug resistance. Pharmacol Ther. 142:316–338. 2014. View Article : Google Scholar : PubMed/NCBI
32	Engelman JA, Zejnullahu K, Mitsudomi T, Song Y, Hyland C, Park JO, Lindeman N, Gale CM, Zhao X, Christensen J, et al: MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling. Science. 316:1039–1043. 2007. View Article : Google Scholar : PubMed/NCBI
33	Karamouzis MV, Konstantinopoulos PA and Papavassiliou AG: Targeting MET as a strategy to overcome crosstalk-related resistance to EGFR inhibitors. Lancet Oncol. 10:709–717. 2009. View Article : Google Scholar : PubMed/NCBI
34	Troiani T, Martinelli E, Napolitano S, Vitagliano D, Ciuffreda LP, Costantino S, Morgillo F, Capasso A, Sforza V, Nappi A, et al: Increased TGF-α as a mechanism of acquired resistance to the anti-EGFR inhibitor cetuximab through EGFR-MET interaction and activation of MET signaling in colon cancer cells. Clin Cancer Res. 19:6751–6765. 2013. View Article : Google Scholar : PubMed/NCBI
35	Luk IY, Reehorst CM and Mariadason JM: ELF3, ELF5, EHF and SPDEF transcription factors in tissue homeostasis and cancer. Molecules. 23:21912018. View Article : Google Scholar : PubMed/NCBI
36	Cheng Z, Guo J, Chen L, Luo N, Yang W and Qu X: Knockdown of EHF inhibited the proliferation, invasion and tumorigenesis of ovarian cancer cells. Mol Carcinog. 55:1048–1059. 2016. View Article : Google Scholar : PubMed/NCBI
37	Fossum SL, Mutolo MJ, Yang R, Dang H, O'Neal WK, Knowles MR, Leir SH and Harris A: Ets homologous factor regulates pathways controlling response to injury in airway epithelial cells. Nucleic Acids Res. 42:13588–13598. 2014. View Article : Google Scholar : PubMed/NCBI