Analysis of naproxen activation of cell death pathways in Colo320 cells

Chen,Andrew; Zhu,Wei; Goding,Rian

doi:10.3892/mco.2024.2759

Analysis of naproxen activation of cell death pathways in Colo320 cells

Authors:
- Andrew Chen
- Wei Zhu
- Rian Goding
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Affiliations: College of Arts and Sciences at The State University of New York at Stony Brook, Stony Brook, NY 11794, USA, SCI Research Institute, Jericho, NY 11753, USA
Published online on: July 3, 2024 https://doi.org/10.3892/mco.2024.2759
Article Number: 61
Copyright: © Chen et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Colorectal cancer is a life‑threatening and prevalent type of cancer. However, a number of current treatments have serious side effects, which increase the need for alternatives. Non‑steroidal anti‑inflammatory drugs have potential chemopreventive capabilities. The present study aimed to confirm this, as well as to investigate potential pathways and reasons for this trait. To accomplish this, cancerous Colo320 and healthy CCD‑18 cells were treated with various concentrations of naproxen sodium (NS). A caspase‑3 assay revealed a statistically significant increase in caspase‑3 activity in Colo320 cells (300%; P<0.01), but not in CCD‑18 cells. This chemical was also associated with a significant decrease in Colo320 cell survival (‑72.888%; P<0.01), but not CCD‑18 cell survival. Furthermore, NS was found to significantly decrease the migration of Colo320 cells (86.58%; P<0.01). Finally, RNA sequencing of cells treated with NS revealed the statistically significant downregulation of the mucin 5B, oligomeric mucus/gel‑forming, S100 calcium binding protein A9 and mucin 5AC, oligomeric mucus/gel‑forming genes, which are upregulated in colorectal cancer and are known to contribute to cancer proliferation, stemness and drug resistance. These novel biological pathway results were further confirmed using ELISAs. The present study identified a novel molecular mechanism of the anti‑colorectal cancer activity of NS.

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1	Centers for Disease Control and Prevention: What is Colorectal Cancer? Centers for Disease Control and Prevention. Available from: https://www.cdc.gov/cancer/colorectal/basic_info/what-is-colorectal-cancer.htm. Accepted at March 19, 2022.
2	American Society of Clinical Oncology. Colorectal Cancer-Types of Treatment. Cancer.Net. Available from: https://www.cancer.net/cancer-types/colorectal-cancer/types-treatment. Accepted at March 19, 2022.
3	Brasky TM, Liu J, White E, Peters U, Potter JD, Walter RB, Baik CS, Lane DS, Manson JAE, Vitolins MZ, et al: Non-steroidal anti-inflammatory drugs and cancer risk in women: Results from the women's health initiative. Int J Cancer. 135:1869–1883. 2014.PubMed/NCBI View Article : Google Scholar
4	Kodela R, Nath N, Chattopadhyay M, Nesbitt DE, Velázquez-Martínez CA and Kashfi K: Hydrogen sulfide-releasing naproxen suppresses colon cancer cell growth and inhibits NF-κB signaling. Drug Des Devel Ther. 9:4873–4882. 2015.PubMed/NCBI View Article : Google Scholar
5	Mohammed A, Janakiram NB, Madka V, Zhang Y, Singh A, Biddick L, Li Q, Lightfoot S, Steele VE, Lubet RA, et al: Intermittent dosing regimens of aspirin and naproxen inhibit azoxymethane-induced colon adenoma progression to adenocarcinoma and invasive carcinoma. Cancer Prev Res (Phila). 12:751–762. 2019.PubMed/NCBI View Article : Google Scholar
6	Reyes-Uribe L, Wu W, Gelincik O, Bommi PV, Francisco-Cruz A, Solis LM, Lynch PM, Lim R, Stoffel EM, Kanth P, et al: Naproxen chemoprevention promotes immune activation in Lynch syndrome colorectal mucosa. Gut. 70:555–566. 2021.PubMed/NCBI View Article : Google Scholar
7	CasPASE™ Apoptosis Assay-G-Biosciences. G-Biosciences. Available from: https://cdn.gbiosciences.com/pdfs/protocol/Caspase_Apoptosis_Colorimentric_Assay_BAQ.pdf. Accepted March 12, 2022.
8	Bumgarner R: Overview of DNA microarrays: Types, applications, and their future. Curr Protoc Mol Biol Chapter 22:Unit 22.1, 2013.
9	Rao MS, Van Vleet TR, Ciurlionis R, Buck WR, Mittelstadt SW, Blomme EAG and Liguori MJ: Comparison of RNA-seq and microarray gene expression platforms for the toxicogenomic evaluation of liver from short-term RAT toxicity studies. Front Genet. 9(636)2019.PubMed/NCBI View Article : Google Scholar
10	Elabscience. Human muc5ac(mucin 5 subtype AC) Elisa Kit. Elabscience. Available from: https://www.elabscience.com/p-human_muc5ac_mucin_5_subtype_ac_elisa_kit-19687.html. Accepted May 6, 2023.
11	Smith CE, Soti S, Jones TA, Nakagawa A, Xue D and Yin H: Non-steroidal anti-inflammatory drugs are caspase inhibitors. Cell Chem Biol. 24:281–292. 2017.PubMed/NCBI View Article : Google Scholar
12	Crowley LC and Waterhouse NJ: Detecting cleaved caspase-3 in apoptotic cells by flow cytometry. Cold Spring Harb Protoc 2016: 2016.
13	Lahdaoui F, Messager M, Vincent A, Hec F, Gandon A, Warlaumont M, Renaud F, Leteurtre E, Piessen G, Jonckheere N, et al: Depletion of MUC5B mucin in gastrointestinal cancer cells alters their tumorigenic properties: Implication of the Wnt/β-catenin pathway. Biochem J. 474:3733–3746. 2017.PubMed/NCBI View Article : Google Scholar
14	Wang Y, Yin K, Tian J, Xia X, Ma J, Tang X, Xu H and Wang S: Granulocytic myeloid-derived suppressor cells promote the stemness of colorectal cancer cells through exosomal S100A9. Adv Sci (Weinh). 6(1901278)2019.PubMed/NCBI View Article : Google Scholar
15	Pothuraju R, Rachagani S, Krishn SR, Chaudhary S, Nimmakayala RK, Siddiqui JA, Ganguly K, Lakshmanan I, Cox JL, Mallya K, et al: Molecular implications of MUC5AC-CD44 axis in colorectal cancer progression and chemoresistance. Mol Cancer. 19(37)2020.PubMed/NCBI View Article : Google Scholar
16	Yuan S, Liu Q, Hu Z, Zhou Z, Wang G, Li C, Xie W, Meng G, Xiang Y, Wu N, et al: Long non-coding RNA MUC5B-AS1 promotes metastasis through mutually regulating MUC5B expression in lung adenocarcinoma. Cell Death Dis. 9(450)2018.PubMed/NCBI View Article : Google Scholar
17	Choi YS, Na HG, Bae CH, Song S and Kim Y: Pepsin exposure in a non-acidic environment upregulates mucin 5AC (MUC5AC) expression via matrix metalloproteinase 9 (mmp9)/nuclear factor ΚB (NF-ΚB) in human airway epithelial cells. Int Forum Allergy Rhinol. 11:894–901. 2021.PubMed/NCBI View Article : Google Scholar
18	Song EJ, Bae CH, Kim JY, Kim YW, Park SY, Song SY and Kim YD: Effect of epigallocatechin-3-gallate on PMA-induced MUC5B expression in human airway epithelial cells. Clin Exp Otorhinolaryngol. 6:237–242. 2013.PubMed/NCBI View Article : Google Scholar
19	Shi S and Yi JL: S100A8/A9 promotes MMP-9 expression in the fibroblasts from cardiac rupture after myocardial infarction by inducing macrophages secreting TNFα. Eur Rev Med Pharmacol Sci. 22:3925–3935. 2018.PubMed/NCBI View Article : Google Scholar
20	Murray D, Morrin M and McDonnell S: Increased invasion and expression of MMP-9 in human colorectal cell lines by a CD44-dependent mechanism. Anticancer Res. 24:489–494. 2004.PubMed/NCBI
21	Espinosa-Cano E, Huerta-Madroñal M, Cámara-Sánchez P, Seras-Franzoso J, Schwartz S Jr, Abasolo I, San Román J and Aguilar MR: Hyaluronic acid (ha)-coated naproxen-nanoparticles selectively target breast cancer stem cells through COX-independent pathways. Mater Sci Eng C Mater Biol Appl. 124(112024)2021.PubMed/NCBI View Article : Google Scholar
22	Kim MS, Kim JE, Lim DY, Huang Z, Chen H, Langfald A, Lubet RA, Grubbs CJ, Dong Z and Bode AM: Naproxen induces cell-cycle arrest and apoptosis in human urinary bladder cancer cell lines and chemically induced cancers by targeting PI3K. Cancer Prev Res (Phila). 7:236–245. 2014.PubMed/NCBI View Article : Google Scholar