Chemosensitizing effect of pentoxifylline in sensitive and multidrug-resistant non-small cell lung cancer cells

Abstract

Aim: Multidrug resistance (MDR) is frequent in non-small cell lung cancer (NSCLC) patients, which can be due to its fibrotic stroma. This work explores the combination of pentoxifylline, an anti-fibrotic and chitinase 3-like-1 (CHI3L1) inhibitor drug, with conventional chemotherapy to improve NSCLC treatment.

Methods: The effect of pentoxifylline in the expression levels of P-glycoprotein (P-gp), CHI3L1 and its main downstream proteins, as well as on cell death, cell cycle profile, and P-gp activity was studied in two pairs of sensitive and MDR counterpart NSCLC cell lines (NCI-H460/NCI-H460/R and A549/A549-CDR2). Association studies between CHI3L1 gene expression and NSCLC patients’ survival were performed using The Cancer Genome Atlas (TCGA) analysis. The sensitizing effect of pentoxifylline to different drug regimens was evaluated in both sensitive and MDR NSCLC cell lines. The cytotoxicity of the drug combinations was assessed in MCF10A non-tumorigenic cells.

Results: Pentoxifylline slightly decreased the expression levels of CHI3L1, β-catenin and signal transducer and activator of transcription 3 (STAT3), and caused a significant increase in the G1 phase of the cell cycle in both pairs of NSCLC cell lines. A significant increase in the % of cell death was observed in the sensitive NCI-H460 cell line. TCGA analysis revealed that high levels of CHI3L1 are associated with low overall survival (OS) in NSCLC patients treated with vinorelbine. Moreover, pentoxifylline sensitized both pairs of sensitive and MDR NSCLC cell lines to the different drug regimens, without causing significant toxicity to non-tumorigenic cells.

Conclusion: This study suggests the possibility of combining pentoxifylline with chemotherapy to increase NSCLC therapeutic response, even in cases of MDR.

Keywords

Chitinase 3-like-1 / combined therapies / drug repurposing / multidrug resistance / non-small cell lung cancer / pentoxifylline

References

[1] Alexander M,Cheng H.Update 2020: management of non-small cell lung cancer.Lung2020;198:897-907 PMCID:PMC7656891

[2] Duma N,Molina JR.Non-small cell lung cancer: epidemiology, screening, diagnosis, and treatment.Mayo Clin Proc2019;94:1623-40

[3] Indini A,Bareggi C.Novel treatment strategies for early-stage lung cancer: the oncologist’s perspective.J Thorac Dis2020;12:3390-8 PMCID:PMC7330760

[4] Lim JU.Update on adjuvant therapy in completely resected NSCLC patients.Thorac Cancer2022;13:277-83 PMCID:PMC8807337

[5] Soon YY,Kim J.Clinical trial and real-world outcomes of patients with metastatic NSCLC in the post-platinum-based chemotherapy failure setting.JTO Clin Res Rep2023;4:100579 PMCID:PMC10628857

[6] Liu WJ,Wen R,Xu J.Drug resistance to targeted therapeutic strategies in non-small cell lung cancer.Pharmacol Ther2020;206:107438

[7] Rotow J.Understanding and targeting resistance mechanisms in NSCLC.Nat Rev Cancer2017;17:637-58

[8] Assaraf YG,Gonçalves AC.The multi-factorial nature of clinical multidrug resistance in cancer.Drug Resist Updat2019;46:100645

[9] Maloney SM,Morejon-Lasso LV.Mechanisms of taxane resistance.Cancers2020;12:3323 PMCID:PMC7697134

[10] Tian Y,Wang Y,Wang J.Mechanism of multidrug resistance to chemotherapy mediated by P-glycoprotein (Review).Int J Oncol2023;63:119 PMCID:PMC10546381

[11] Wang C,Sun B.Hsa-miR-4271 downregulates the expression of constitutive androstane receptor and enhances in vivo the sensitivity of non-small cell lung cancer to gefitinib.Pharmacol Res2020;161:105110

[12] Chen C,Yu S.Role of cancer-associated fibroblasts in the resistance to antitumor therapy, and their potential therapeutic mechanisms in non-small cell lung cancer.Oncol Lett2021;21:413 PMCID:PMC8020389

[13] Han X,Geranpayehvaghei M,Li Y.Emerging nanomedicines for anti-stromal therapy against desmoplastic tumors.Biomaterials2020;232:119745

[14] Zhang H,Zhu L,Ma S.Cancer-associated fibroblasts in non-small cell lung cancer: recent advances and future perspectives.Cancer Lett2021;514:38-47

[15] Lin Y,Zhou B,Jiang M.Pentoxifylline inhibits pulmonary fibrosis by regulating cellular senescence in mice.Front Pharmacol2022;13:848263 PMCID:PMC9160723

[16] Wen WX,Siang R.Repurposing pentoxifylline for the treatment of fibrosis: an overview.Adv Ther2017;34:1245-69

[17] Xavier CPR,Caires HR.Chitinase 3-like-1 and fibronectin in the cargo of extracellular vesicles shed by human macrophages influence pancreatic cancer cellular response to gemcitabine.Cancer Lett2021;501:210-23

[18] Bravo-Cuellar A,Lerma-Diaz JM.Sensitization of cervix cancer cells to adriamycin by pentoxifylline induces an increase in apoptosis and decrease senescence.Mol Cancer2010;9:114 PMCID:PMC2890603

[19] Bravo-Cuellar A,Sierra-Díaz E.Pentoxifylline sensitizes cisplatin-resistant human cervical cancer cells to cisplatin treatment: involvement of mitochondrial and NF-kappa B pathways.Front Oncol2020;10:592706 PMCID:PMC7931705

[20] Cancino-Marentes ME,Ortiz-Lazareno PC.Sensitizing the cytotoxic action of docetaxel induced by pentoxifylline in a PC3 prostate cancer cell line.BMC Urol2021;21:38 PMCID:PMC7953714

[21] Kim JH,Park WS,Kuh HJ.Antifibrotic effects of pentoxifylline improve the efficacy of gemcitabine in human pancreatic tumor xenografts.Cancer Sci2017;108:2470-7 PMCID:PMC5715266

[22] Ohsaki Y,Fujikane T.Pentoxifylline potentiates the antitumor effect of cisplatin and etoposide on human lung cancer cell lines.Oncology1996;53:327-33

[23] Goel PN.Curbing the focal adhesion kinase and its associated signaling events by pentoxifylline in MDA-MB-231 human breast cancer cells.Eur J Pharmacol2013;714:432-41

[24] Goel PN.Unravelling the antimetastatic potential of pentoxifylline, a methylxanthine derivative in human MDA-MB-231 breast cancer cells.Mol Cell Biochem2011;358:141-51

[25] Wang Y,Li J,Shang B.Pentoxifylline induces apoptosis of HepG2 cells by reducing reactive oxygen species production and activating the MAPK signaling.Life Sci2017;183:60-8

[26] Al-husein BA,Alzoubi KH.Pentoxifylline induces caspase-dependent apoptosis in colorectal cancer cells.Inform Med Unlocked2022;31:100997

[27] Rao FV,Vora KA,van Aalten DM.Methylxanthine drugs are chitinase inhibitors: investigation of inhibition and binding modes.Chem Biol2005;12:973-80

[28] Chen HT,Zhang YZ.Overexpression of YKL-40 predicts poor prognosis in patients undergoing curative resection of pancreatic cancer.Pancreas2017;46:323-34

[29] De Robertis M,Cardone RA.Upregulation of YKL-40 promotes metastatic phenotype and correlates with poor prognosis and therapy response in patients with colorectal cancer.Cells2022;11:3568 PMCID:PMC9688424

[30] Lee YE,Tian YF.High expression of chitinase 3-like-1 is an unfavorable prognostic factor in urothelial carcinoma of upper urinary tract and urinary bladder.Urol Oncol2019;37:299.e7-18

[31] Jefri M,Huang WC,Chen WL.YKL-40 regulated epithelial-mesenchymal transition and migration/invasion enhancement in non-small cell lung cancer.BMC Cancer2015;15:590 PMCID:PMC4537570

[32] Park KR,Yoo K,Han SB.Chitinase 3 like 1 suppresses the stability and activity of p53 to promote lung tumorigenesis.Cell Commun Signal2020;18:5 PMCID:PMC7055043

[33] Zhao T,Li Y,You Q.Chitinase-3 like-protein-1 function and its role in diseases.Signal Transduct Target Ther2020;5:201 PMCID:PMC7490424

[34] Geng B,Zhao T.Chitinase 3-like 1-CD44 interaction promotes metastasis and epithelial-to-mesenchymal transition through β-catenin/Erk/Akt signaling in gastric cancer.J Exp Clin Cancer Res2018;37:208 PMCID:PMC6117920

[35] Jaén M,Bartolomé RA,Casal JI.Interleukin 13 receptor alpha 2 (IL13Rα2): expression, signaling pathways and therapeutic applications in cancer.Biochim Biophys Acta Rev Cancer2022;1877:188802

[36] Yu JE,Han SB.Significance of chitinase-3-like protein 1 in the pathogenesis of inflammatory diseases and cancer.Exp Mol Med2024;56:1-18 PMCID:PMC10834487

[37] Pesic M,Jankovic D.Induced resistance in the human non small cell lung carcinoma (NCI-H460) cell line in vitro by anticancer drugs.J Chemother2006;18:66-73

[38] Podolski-Renić A,Stanković T.Molecular and cytogenetic changes in multi-drug resistant cancer cells and their influence on new compounds testing.Cancer Chemother Pharmacol2013;72:683-97

[39] Silva BR,Rodrigues JM,Vasconcelos MH.Synthesis of novel methyl 3-(hetero)arylthieno[3,2-b]pyridine-2-carboxylates and antitumor activity evaluation: studies in vitro and in ovo grafts of chick chorioallantoic membrane (CAM) with a triple negative breast cancer cell line.Molecules2021;26:1594 PMCID:PMC7999514

[40] Vichai V.Sulforhodamine B colorimetric assay for cytotoxicity screening.Nat Protoc2006;1:1112-6

[41] Branco H,Antunes C,Vasconcelos MH.Pirfenidone sensitizes NCI-H460 non-small cell lung cancer cells to paclitaxel and to a combination of paclitaxel with carboplatin.Int J Mol Sci2022;23:3631 PMCID:PMC8998757

[42] Teixeira A,Barros L.Eucalyptus globulus Labill. decoction extract inhibits the growth of NCI-H460 cells by increasing the p53 levels and altering the cell cycle profile.Food Funct2019;10:3188-97

[43] Castro I,Branco H,Xavier CPR.Establishing and characterizing a novel doxorubicin-resistant acute myeloid leukaemia cell line.J Chemother2023;35:307-21

[44] Lopes-Rodrigues V,Correia-da-Silva M.A novel curcumin derivative which inhibits P-glycoprotein, arrests cell cycle and induces apoptosis in multidrug resistance cells.Bioorg Med Chem2017;25:581-96

[45] Lopes-Rodrigues V,Mleczko J.Identification of the metabolic alterations associated with the multidrug resistant phenotype in cancer and their intercellular transfer mediated by extracellular vesicles.Sci Rep2017;7:44541 PMCID:PMC5356019

[46] Lopes-Rodrigues V,Sousa D.Multidrug resistant tumour cells shed more microvesicle-like EVs and less exosomes than their drug-sensitive counterpart cells.Biochim Biophys Acta2016;1860:618-27

[47] Bian B,Yang J.Prognostic value of YKL-40 in solid tumors: a meta-analysis of 41 cohort studies.Cancer Cell Int2019;19:259 PMCID:PMC6785874

[48] Barancik M,Gibalova L,Sulova Z.Potentiation of anticancer drugs: effects of pentoxifylline on neoplastic cells.Int J Mol Sci2012;13:369-82 PMCID:PMC3269692

[49] Drobná Z,Walther W,Breier A.Pentoxifylline influences drug transport activity of P-glycoprotein and decreases mdrl gene expression in multidrug resistant mouse leukemic L1210/VCR cells.Gen Physiol Biophys2002;21:103-9

[50] Feng C,Zhang S,Li C.Therapeutic effects of pentoxifylline on invasive pulmonary aspergillosis in immunosuppressed mice.BMC Pulm Med2021;21:31 PMCID:PMC7814429

[51] Gude RP,Presas HL,Bonfil RD.Studies on the mechanisms responsible for inhibition of experimental metastasis of B16-F10 murine melanoma by pentoxifylline.J Biomed Sci1999;6:133-41

[52] Kamran MZ.Pentoxifylline inhibits melanoma tumor growth and angiogenesis by targeting STAT3 signaling pathway.Biomed Pharmacother2013;67:399-405

[53] Park KR,Hong JT.G721-0282 inhibits cell growth and induces apoptosis in human osteosarcoma through down-regulation of the STAT3 pathway.Int J Biol Sci2020;16:330-41 PMCID:PMC6949149

[54] Lee DH,Hwang CJ.Decreased lung tumor development in SwAPP mice through the downregulation of CHI3L1 and STAT 3 activity via the upregulation of miRNA342-3p.Mol Ther Nucleic Acids2019;16:63-72 PMCID:PMC6406047

[55] Singh SK,Wilczynska KM,Kordula T.A complex of nuclear factor I-X3 and STAT3 regulates astrocyte and glioma migration through the secreted glycoprotein YKL-40.J Biol Chem2011;286:39893-903 PMCID:PMC3220556

[56] Yan C,Wu L,Qu P.Stat3 downstream gene product chitinase 3-like 1 is a potential biomarker of inflammation-induced lung cancer in multiple mouse lung tumor models and humans.PLoS One2013;8:e61984 PMCID:PMC3632574

[57] Castellanos-Esparza YC,Huang L.Synergistic promoting effects of pentoxifylline and simvastatin on the apoptosis of triple-negative MDA-MB-231 breast cancer cells.Int J Oncol2018;52:1246-54

[58] Yeo IJ,Han SB,Hong JT.Roles of chitinase 3-like 1 in the development of cancer, neurodegenerative diseases, and inflammatory diseases.Pharmacol Ther2019;203:107394

[59] Chiang YC,Chang CF.Overexpression of CHI3L1 is associated with chemoresistance and poor outcome of epithelial ovarian carcinoma.Oncotarget2015;6:39740-55 PMCID:PMC4741859

[60] Gong M,Ye X.Loss-of-function mutations in KEAP1 drive lung cancer progression via KEAP1/NRF2 pathway activation.Cell Commun Signal2020;18:98 PMCID:PMC7310414

[61] Adinolfi S,Jawahar Deen A.The KEAP1-NRF2 pathway: targets for therapy and role in cancer.Redox Biol2023;63:102726 PMCID:PMC10189287

[62] Telkoparan-Akillilar P,Cevik D,Saso L.Therapeutic targeting of the NRF2 signaling pathway in cancer.Molecules2021;26:1417 PMCID:PMC7961421

[63] Sadeghiani G,Sadighi Gilani MA,Malekzadeh M.Evaluation of Nrf2/ARE signaling pathway in the presence of pentoxifylline as a cryoprotectant in mouse spermatogonial stem cells.Biopreserv Biobank2023;21:294-307