Gene expression alterations in hypoxic A549 lung cancer cell line

Abualnadi,Rania; Tarboush,Nafez Abu; Shhab,Mohammad; Zihlif,Malek

doi:10.3892/br.2024.1871

Gene expression alterations in hypoxic A549 lung cancer cell line

Authors:
- Rania Abualnadi
- Nafez Abu Tarboush
- Mohammad Shhab
- Malek Zihlif
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Affiliations: Department of Pharmacology, School of Medicine, The University of Jordan, Amman 11942, Jordan, Department of Physiology and Biochemistry, School of Medicine, The University of Jordan, Amman 11942, Jordan
Published online on: October 4, 2024 https://doi.org/10.3892/br.2024.1871
Article Number: 183
Copyright: © Abualnadi et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Human non‑small cell lung cancer (NSCLC)is a very common disease with limited treatment options. Hypoxia is a characteristic feature of solid tumors associated with the resistance of cancer cells to radiotherapy and chemotherapy. Therefore, the expression changes in cancer‑resistance genes may be biomarkers of hypoxia with value in targeted therapy. The aim of the present study was to examine the effect of hypoxia on gene expression and the changes that occur in relation to drug resistance in a human NSCLC cell line (A549). A549 cells were exposed to 72‑h hypoxic episodes (<1% oxygen) for a total of 10 episodes (acute). The alterations in gene expression were examined using PCR array technology after 10 episodes of acute hypoxia and compared with normoxic cells. The chemoresistance of hypoxic cells toward doxorubicin was measured using a MTT cell proliferation assay. A549 cells were affected by acute hypoxia leading to induced doxorubicin chemoresistance. Evident changes in the gene expression level were identified following episodes of acute hypoxia. The most important changes occurred in the estrogen receptor 1 (ESR1) and Finkel‑Biskis‑Jinkins osteosarcoma (FOS) pathways and in different nucleic transcription factors such as aryl hydrocarbon receptor and cyclin‑dependent kinase inhibitor. The present study showed that exposing cells to prolonged periods of hypoxia results in different gene expression changes. There was induction of chemo‑resistance due to acute hypoxia. ESR1 and c‑FOS are proposed as a potential hypoxia genes in lung cancer.

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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.PubMed/NCBI View Article : Google Scholar
2	Jemal A, Siegel R, Ward E, Hao Y, Xu J, Murray T and Thun MJ: Cancer Statistics, 2008. CA Cancer J Clin. 58:71–96. 2008.PubMed/NCBI View Article : Google Scholar
3	Sinha BK, Mimnaugh EG, Rajagopalan S and Myers CE: Adriamycin activation and oxygen free radical formation in human breast tumor cells: Protective role of glutathione peroxidase in adriamycin resistance 1. Cancer Res. 49:3844–3848. 1989.PubMed/NCBI
4	Vaupel P and Harrison L: Tumor hypoxia: Causative factors, compensatory mechanisms, and cellular response. Oncologist. 9 (Suppl 5):S4–S9. 2004.PubMed/NCBI View Article : Google Scholar
5	Das B, Tsuchida R, Malkin D, Koren G, Baruchel S and Yeger H: Hypoxia enhances tumor stemness by increasing the invasive and tumorigenic side population fraction. Stem Cells. 26:1818–1830. 2008.PubMed/NCBI View Article : Google Scholar
6	Chen Z, Han F, Du Y, Shi H and Zhou W: Hypoxic microenvironment in cancer: Molecular mechanisms and therapeutic interventions. Sig Transduct Target Ther. 8(70)2023.PubMed/NCBI View Article : Google Scholar
7	Bayer C and Vaupel P: Acute versus chronic hypoxia in tumors : Controversial data concerning time frames and biological consequences. Strahlenther Onkol. 188:616–627. 2012.PubMed/NCBI View Article : Google Scholar
8	Le QT, Chen E, Salim A, Cao H, Kong CS, Whyte R, Donington J, Cannon WA, Wakelee H, Tibshirani R, et al: An evaluation of tumor oxygenation and gene expression in patients with early stage non-small cell lung cancers. Clin. Cancer Res. 12:1507–1514. 2006.PubMed/NCBI View Article : Google Scholar
9	Ziółkowska-Suchanek I: Mimicking tumor hypoxia in non-small cell lung cancer employing three-dimensional in vitro models. Cells. 10(141)2021.PubMed/NCBI View Article : Google Scholar
10	Marhuenda E, Campillo N, Gabasa M, Martínez-García MA, Campos-Rodríguez F, Gozal D, Navajas D, Alcaraz J, Farré R and Almendros I: Effects of Sustained and Intermittent Hypoxia on Human Lung Cancer Cells. Am J Respir Cell Mol Biol. 61:540–544. 2019.PubMed/NCBI View Article : Google Scholar
11	Rouschop KM, Ramaekers CH, Schaaf MB, Keulers TG, Savelkouls KG, Lambin P, Koritzinsky M and Wouters BG: Autophagy is required during cycling hypoxia to lower production of reactive oxygen species. Radiother Oncol. 92:411–416. 2009.PubMed/NCBI View Article : Google Scholar
12	Hsieh CH, Shyu WC, Chiang CY, Kuo JW, Shen WC and Liu RS: NADPH oxidase subunit 4-mediated reactive oxygen species contribute to cycling hypoxia-promoted tumor progression in glioblastoma multiforme. PLoS One. 6(e23945)2011.PubMed/NCBI View Article : Google Scholar
13	Rofstad EK, Gaustad JV, Egeland TA, Mathiesen B and Galappathi K: Tumors exposed to acute cyclic hypoxic stress show enhanced angiogenesis, perfusion and metastatic dissemination. Int J Cancer. 127:1535–1546. 2010.PubMed/NCBI View Article : Google Scholar
14	Pires IM, Bencokova Z, Milani M, Folkes LK, Li JL, Stratford MR, Harris AL and Hammond EM: Effects of acute versus chronic hypoxia on DNA damage responses and genomic instability. Cancer Res. 70:925–935. 2010.PubMed/NCBI View Article : Google Scholar
15	Erler JT, Cawthorne CJ, Williams KJ, Koritzinsky M, Wouters BG, Wilson C, Miller C, Demonacos C, Stratford IJ and Dive C: Hypoxia-Mediated Down-Regulation of Bid and Bax in Tumors Occurs via Hypoxia-Inducible Factor 1-Dependent and -Independent Mechanisms and Contributes to Drug Resistance. Mol Cell Biol. 24:2875–2889. 2004.PubMed/NCBI View Article : Google Scholar
16	Vaupel P, Kelleher DK and Höckel M: Oxygenation status of malignant tumors: Pathogenesis of hypoxia and significance for tumor therapy. Semin Oncol. 28 (2 Suppl 8):29–35. 2001.PubMed/NCBI View Article : Google Scholar
17	Al-Najjar LM, Zihlif M and Jarrar Y: Differential molecular alterations promoting non-small cell lungcancer under hypoxia. Genome Instab Dis. 3:108–121. 2022.
18	Chen YL, Yang TY, Chen KC, Wu CL, Hsu SL and Hsueh CM: Hypoxia can impair doxorubicin resistance of non-small cell lung cancer cells by inhibiting MRP1 and P-gp expression and boosting the chemosensitizing effects of MRP1 and P-gp blockers. Cell Oncol (Dordr). 39:411–433. 2016.PubMed/NCBI View Article : Google Scholar
19	Chou CW, Wang CC, Wu CP, Lin YJ, Lee YC, Cheng YW and Hsieh CH: Tumor cycling hypoxia induces chemoresistance in glioblastoma multiforme by upregulating the expression and function of ABCB1. Neuro Oncol. 14:1227–1238. 2012.PubMed/NCBI View Article : Google Scholar
20	Green SL and Giaccia AJ: Tumor hypoxia and the cell cycle: Implications for malignant progression and response to therapy. Cancer J Sci Am. 4:218–223. 1998.PubMed/NCBI
21	Zhou SF, Wang B, Yang LP and Liu JP: Structure, function, regulation and polymorphism and the clinical significance of human cytochrome P450 1A2. Drug Metab Rev. 42:268–354. 2010.PubMed/NCBI View Article : Google Scholar
22	AbuHammad S and Zihlif M: Gene expression alterations in doxorubicin resistant MCF7 breast cancer cell line. Genomics. 101:213–220. 2013.PubMed/NCBI View Article : Google Scholar
23	Lei JT, Gou X, Seker S and Ellis MJ: ESR1 alterations and metastasis in estrogen receptor positive breast cancer. J Cancer Metastasis Treat. 5(38)2019.PubMed/NCBI View Article : Google Scholar
24	Fialka I, Schwarz H, Reichmann E, Oft M, Busslinger M and Beug H: The estrogen-dependent C-junER protein causes a reversible loss of mammary epithelial cell polarity involving a destabilization of adherens junctions. J Cell Biol. 132:1115–1132. 1996.PubMed/NCBI View Article : Google Scholar
25	Gartel AL and Tyner AL: The role of the cyclin-dependent kinase inhibitor p21 in apoptosis. Mol Cancer Ther. 1:639–649. 2002.PubMed/NCBI
26	Bunz F, Hwang PM, Torrance C, Waldman T, Zhang Y, Dillehay L, Williams J, Lengauer C, Kinzler KW and Vogelstein B: Disruption of p53 in human cancer cells alters the responses to therapeutic agents. J Clin. Invest. 104:263–269. 1999.PubMed/NCBI View Article : Google Scholar
27	Muhammad N, Bhattacharya S, Steele R, Phillips N and Ray RB: Involvement of c-Fos in the promotion of cancer stem-like cell properties in head and neck squamous cell carcinoma. Clin Cancer Res. 23:3120–3128. 2017.PubMed/NCBI View Article : Google Scholar
28	Sermeus A, Genin M, Maincent A, Fransolet M, Notte A, Leclere L, Riquier H, Arnould T and Michiels C: Hypoxia-Induced modulation of apoptosis and BCL-2 family proteins in different cancer cell types. PLoS One. 7(e47519)2012.PubMed/NCBI View Article : Google Scholar
29	Adams JM and Cory S: The Bcl-2 apoptotic switch in cancer development and therapy. Oncogene. 26:1324–1337. 2007.PubMed/NCBI View Article : Google Scholar
30	Brtko J: Role of retinoids and their cognate nuclear receptors in breast cancer chemoprevention. Cent Eur J Public Health. 15:3–6. 2007.PubMed/NCBI View Article : Google Scholar
31	Hua F, Fang N, Li X, Zhu S, Zhang W and Gu J: A meta-analysis of the relationship between RARβ gene promoter methylation and non-small cell lung cancer. PLoS One. 9(e96163)2014.PubMed/NCBI View Article : Google Scholar
32	Liu Z, Zhang L, Ding F, Li J, Guo M, Li W, Wang Y, Yu Z, Zhan Q, Wu M and Liu Z: 5-Aza-2'-deoxycytidine induces retinoic acid receptor-beta(2) demethylation and growth inhibition in esophageal squamous carcinoma cells. Cancer Lett. 230:271–283. 2005.PubMed/NCBI View Article : Google Scholar
33	Pérez RJ, Benoit YD and Gudas LJ: Deletion of retinoic acid receptor β (RARβ) impairs pancreatic endocrine differentiation. Exp Cell Res. 319:2196–204. 2013.PubMed/NCBI View Article : Google Scholar
34	Ren HY, Chen B, Huang GL, Liu Y and Shen DY: Upregulation of retinoic acid receptor-β reverses drug resistance in cholangiocarcinoma cells by enhancing susceptibility to apoptosis. Mol Med Rep. 14:3602–3608. 2016.PubMed/NCBI View Article : Google Scholar
35	Li Y, Lu DG, Ma YM and Liu H: Association between Retinoic acid receptor-β hypermethylation and NSCLC risk: A meta-analysis and literature review. Oncotarget. 8:5814–5822. 2017.PubMed/NCBI View Article : Google Scholar
36	Demchenko YN, Glebov OK, Zingone A, Keats JJ, Bergsagel PL and Kuehl WM: Classical and/or alternative NF-kappaB pathway activation in multiple myeloma. Blood. 115:3541–3552. 2010.PubMed/NCBI View Article : Google Scholar
37	Qin H, Zhou J, Zhou P, Xu J, Tang Z, Ma H and Guo F: Prognostic significance of RelB overexpression in non-small cell lung cancer patients. Thorac. Cancer. 7:415–421. 2016.PubMed/NCBI View Article : Google Scholar
38	Adam AP, George A, Schewe D, Bragado P, Iglesias BV, Ranganathan AC, Kourtidis A, Conklin DS and Aguirre-Ghiso JA: Computational identification of a p38SAPK-regulated transcription factor network required for tumor cell quiescence. Cancer Res. 69:5664–5672. 2009.PubMed/NCBI View Article : Google Scholar
39	Zhao Y, Lu H, Yan A, Yang Y, Meng Q, Sun L, Pang H, Li C, Dong X and Cai L: ABCC3 as a marker for multidrug resistance in non-small cell lung cancer. Sci Rep. 3(3120)2013.PubMed/NCBI View Article : Google Scholar
40	Fukuda Y and Schuetz JD: ABC transporters and their role in nucleoside and nucleotide drug resistance. Biochem Pharmacol. 83:1073–1083. 2012.PubMed/NCBI View Article : Google Scholar
41	Kirovski G, Stevens AP, Czech B, Dettmer K, Weiss TS, Wild P, Hartmann A, Bosserhoff AK, Oefner PJ and Hellerbrand C: Down-regulation of methylthioadenosine phosphorylase (MTAP) induces progression of hepatocellular carcinoma via accumulation of 5'-deoxy-5'-methylthioadenosine (MTA). Am J Pathol. 178:1145–1152. 2011.PubMed/NCBI View Article : Google Scholar
42	Chu IM, Hengst L and Slingerland JM: The Cdk inhibitor p27 in human cancer: Prognostic potential and relevance to anticancer therapy. Nat Rev Cancer. 8:253–267. 2008.PubMed/NCBI View Article : Google Scholar
43	Semenza GL: Intratumoral hypoxia and mechanisms ofimmune evasion mediated by hypoxia-inducible factors. Physiology (Bethesda). 36:73–83. 2021.PubMed/NCBI View Article : Google Scholar