Butein-instigated miR-186-5p-dependent modulation of TWIST1 affects resistance to cisplatin and bioenergetics of Malignant Pleural Mesothelioma cells

Abstract

Aim: Malignant pleural mesothelioma is a chemoresistant tumor, and biphasic and sarcomatoid histologies portend the worst prognosis for malignant pleural mesothelioma (MPM) patients. We obtained the microRNA expression profile of three biphasic-sarcomatoid MPM cell lines to identify commonly expressed microRNAs and evaluate the effect of butein, a chemo-sensitizing compound, on this microRNA subset.

Methods: Nanostring-based microRNA profiling and analysis through the ROSALIND platform were employed to identify the commonly modulated microRNAs and their targets. MicroRNA-mimic transfection, Luciferase assay, and Western blotting were employed to show specific perturbation of TWIST1 levels by miR-186-5p. Sphere-forming assays, invasion assay, and metabolic profiling were used to assess the biological consequences of the butein-instigated miR-186-5p-mediated perturbation of TWIST1 levels. TGCA analysis was used to search for the correlation between TWIST1 and miR-186-5p levels in biphasic and epithelioid MPM specimens.

Results: We identified a set of perturbed microRNAs, common to three biphasic/sarcomatoid MPM cell lines, after butein treatment. When focusing on miR-186-5p, we unraveled a butein-ignited and miR-186-5p-mediated modulation of TWIST1 levels which affected the 3D anchorage-independent growth, cisplatin resistance, invasion, and bioenergetics of the MPM cell lines tested. We showed that miR-186-5p and TWIST1 levels are anti-correlated in biphasic MPM specimens from TCGA.

Conclusion: We unraveled a novel mechanism of action of butein, which attenuated the pro-tumorigenic features of MPM at least through a miR-186-5p-TWIST1 axis. We suggest that those activities converge into the chemo-sensitizing effect of this compound and may be of translational relevance.

Keywords

Mesothelioma / butein / miR-186-5p / TWIST1 / pithelial-to-mesenchymal transition (EMT) / chemoresistance / cancer metabolism / invasion / anchorage-independent growth

References

[1] Asciak R,Rahman NM.Update on biology and management of mesothelioma.Eur Respir Rev2021;30:200226

[2] Hiltbrunner S,Kirschner MB.Tumor immune microenvironment and genetic alterations in mesothelioma.Front Oncol2021;11:660039 PMCID:PMC8261295

[3] Brcic L.Clinical significance of histologic subtyping of malignant pleural mesothelioma.Transl Lung Cancer Res2020;9:924-33 PMCID:PMC7354152

[4] Blum Y,Quetel L.Dissecting heterogeneity in malignant pleural mesothelioma through histo-molecular gradients for clinical applications.Nat Commun2019;10:1333 PMCID:PMC6430832

[5] Ramundo V,Aldieri E.The epithelial-to-mesenchymal transition (EMT) in the development and metastasis of malignant pleural mesothelioma.Int J Mol Sci2021;22:12216 PMCID:PMC8621591

[6] Iwanami T,Nakagawa M.Clinical significance of epithelial-mesenchymal transition-associated markers in malignant pleural mesothelioma.Oncology2014;86:109-16

[7] Nicolini F,Bronte G.Malignant Pleural mesothelioma: state-of-the-art on current therapies and promises for the future.Front Oncol2019;9:1519 PMCID:PMC6992646

[8] Baas P,Lacoin L.Treatment patterns and outcomes for patients with malignant pleural mesothelioma in England in 2013-2017: a nationwide CAS registry analysis from the I-O Optimise initiative.Lung Cancer2021;162:185-93

[9] Meirson T,Cerza F.Comparison of 3 randomized clinical trials of frontline therapies for malignant pleural mesothelioma.JAMA Netw Open2022;5:e221490 PMCID:PMC8908075

[10] Mujoomdar AA,Richards WG,Sugarbaker DJ.Prevalence of in vitro chemotherapeutic drug resistance in primary malignant pleural mesothelioma: result in a cohort of 203 resection specimens.J Thorac Cardiovasc Surg2010;140:352-5

[11] Canino C,Cambria A.SASP mediates chemoresistance and tumor-initiating-activity of mesothelioma cells.Oncogene2012;31:3148-63

[12] Cioce M,Pass H,Strano S.Arachidonic acid drives adaptive responses to chemotherapy-induced stress in malignant mesothelioma.J Exp Clin Cancer Res2021;40:344 PMCID:PMC8561918

[13] di Martino S,Nuvoli B.HSP90 inhibition alters the chemotherapy-driven rearrangement of the oncogenic secretome.Oncogene2018;37:1369-85

[14] Manfredi M,Gosetti F,Marengo E.The secretome signature of malignant mesothelioma cell lines.J Proteomics2016;145:3-10

[15] Canino C,Marcato P,Pass HI.A STAT3-NFkB/DDIT3/CEBPβ axis modulates ALDH1A3 expression in chemoresistant cell subpopulations.Oncotarget2015;6:12637-53 PMCID:PMC4494963

[16] Cioce M,Pulito C,Strano S.Butein impairs the protumorigenic activity of malignant pleural mesothelioma cells.Cell Cycle2012;11:132-40

[17] Cioce M,Pass HI.Insights into intra-tumoral heterogeneity: transcriptional profiling of chemoresistant MPM cell subpopulations reveals involvement of NFkB and DNA repair pathways and contributes a prognostic signature.Int J Mol Sci2021;22:12071 PMCID:PMC8585077

[18] Jayasooriya RGPT,Park C.Molecular chemotherapeutic potential of butein: a concise review.Food Chem Toxicol2018;112:1-10

[19] Moon DO,Lee JD,Kim GY.Butein suppresses c-Myc-dependent transcription and Akt-dependent phosphorylation of hTERT in human leukemia cells.Cancer Lett2009;286:172-9

[20] Zhang L,Li X.A novel anticancer effect of butein: inhibition of invasion through the ERK1/2 and NF-kappa B signaling pathways in bladder cancer cells.FEBS Lett2008;582:1821-8

[21] He L.MicroRNAs: small RNAs with a big role in gene regulation.Nat Rev Genet2004;5:522-31

[22] Meijer HA,Lu WT.Translational repression and eIF4A2 activity are critical for microRNA-mediated gene regulation.Science2013;340:82-5

[23] Doldi V,Zaffaroni N.MicroRNAs as epigenetic determinants of treatment response and potential therapeutic targets in prostate cancer.Cancers (Basel)2021;13:2380 PMCID:PMC8156532

[24] Moghbeli M.MicroRNAs as the critical regulators of Cisplatin resistance in ovarian cancer cells.J Ovarian Res2021;14:127 PMCID:PMC8485521

[25] Pouya FD,Gazouli M,Nemati M.MicroRNAs as therapeutic targets in breast cancer metastasis.Drug Deliv Transl Res2022;12:1029-46

[26] Semina EV,Troyanovskiy KE,Rubina KA.MicroRNAs in cancer: from gene expression regulation to the metastatic niche reprogramming.Biochemistry (Mosc)2021;86:785-99

[27] Xiang Y,Guan L.The dual role of miR-186 in cancers: oncomir battling with tumor suppressor miRNA.Front Oncol2020;10:233 PMCID:PMC7066114

[28] Zhang Z,Mao J,Fan M.miR-186-5p functions as a tumor suppressor in human osteosarcoma by targeting FOXK1.Cell Physiol Biochem2019;52:553-64

[29] Ye J,Sun L,Xu X.miR-186 regulates chemo-sensitivity to paclitaxel via targeting MAPT in non-small cell lung cancer (NSCLC).Mol Biosyst2016;12:3417-24

[30] Sun WJ,Xue P.miR-186 inhibits proliferation, migration, and epithelial-mesenchymal transition in breast cancer cells by targeting Twist1.J Cell Biochem2019;120:10001-9

[31] Liu X,Chen Y,He J.miR-186-5p targeting SIX1 inhibits cisplatin resistance in non-small-cell lung cancer cells (NSCLCs).Neoplasma2020;67:147-57

[32] Li J,Guo F.miR-186 reverses cisplatin resistance and inhibits the formation of the glioblastoma-initiating cell phenotype by degrading Yin Yang 1 in glioblastoma.Int J Mol Med2019;43:517-24

[33] Li C,Li Y.TUG1 mediates methotrexate resistance in colorectal cancer via miR-186/CPEB2 axis.Biochem Biophys Res Commun2017;491:552-7

[34] Cao C,Zhang L.miR-186 affects the proliferation, invasion and migration of human gastric cancer by inhibition of Twist1.Oncotarget2016;7:79956-63 PMCID:PMC5346763

[35] Wang XX,Zhang ZH.TWIST1 transcriptionally regulates glycolytic genes to promote the Warburg metabolism in pancreatic cancer.Exp Cell Res2020;386:111713

[36] Fischer KR,Lee S.Epithelial-to-mesenchymal transition is not required for lung metastasis but contributes to chemoresistance.Nature2015;5274:472-6 PMCID:PMC4662610

[37] Cioce M.On the connections between cancer stem cells and EMT.Cell Cycle2012;11:4301-2 PMCID:PMC3552909

[38] Merikallio H,Salmenkivi K,Harju T.Expression of snail, twist, and Zeb1 in malignant mesothelioma.APMIS2013;121:1-10

[39] Tan Z,Yan CW.Antimesothelioma Immunotherapy by CTLA-4 blockade depends on active PD1-based TWIST1 vaccination.Mol Ther Oncolytics2020;16:302-17 PMCID:PMC7068049

[40] Fassina A,Guzzardo V.Epithelial-mesenchymal transition in malignant mesothelioma.Mod Pathol2012;25:86-99

[41] Schramm A,Thies S.Prognostic significance of epithelial-mesenchymal transition in malignant pleural mesothelioma.Eur J Cardiothorac Surg2010;37:566-72

[42] Cortes-Dericks L,Boesch R,Karoubi G.Cisplatin-resistant cells in malignant pleural mesothelioma cell lines show ALDH^highCD⁴⁴⁺ phenotype and sphere-forming capacity.BMC Cancer2014;14:304 PMCID:PMC4021184

[43] Pasdar EA,Stapelberg M.Characterisation of mesothelioma-initiating cells and their susceptibility to anti-cancer agents.PLoS ONE2015;10:e0119549 PMCID:PMC4416766

[44] Baas P,Nowak AK.First-line nivolumab plus ipilimumab in unresectable malignant pleural mesothelioma (CheckMate 743): a multicentre, randomised, open-label, phase 3 trial.The Lancet2021;397:375-86

[45] Del Peso G,Bajo MA.Epithelial-to-mesenchymal transition of mesothelial cells is an early event during peritoneal dialysis and is associated with high peritoneal transport.Kidney Int Suppl2008;:S26-33

[46] Aroeira LS,Sánchez-Tomero JA.Epithelial to mesenchymal transition and peritoneal membrane failure in peritoneal dialysis patients: pathologic significance and potential therapeutic interventions.J Am Soc Nephrol2007;18:2004-13

[47] Li Z,Shen J,Chan MT.MicroRNA dysregulation in rhabdomyosarcoma: a new player enters the game.Cell Prolif2015;48:511-6 PMCID:PMC6496529

[48] Bahar E,Kim DC,Yoon H.Combination of Niraparib, cisplatin and twist knockdown in cisplatin-resistant ovarian cancer cells potentially enhances synthetic lethality through er-stress mediated mitochondrial apoptosis pathway.Int J Mol Sci2021;22:3916 PMCID:PMC8070209

[49] Wu YH,Chang TH.Activation of TWIST1 by COL11A1 promotes chemoresistance and inhibits apoptosis in ovarian cancer cells by modulating NF-κB-mediated IKKβ expression.Int J Cancer2017;141:2305-17

[50] Roberts CM,Pitruzzello MC.TWIST1 drives cisplatin resistance and cell survival in an ovarian cancer model, via upregulation of GAS6, L1CAM, and Akt signalling.Sci Rep2016;6:37652 PMCID:PMC5120297

[51] Dietze R,Gómez-Serrano M.Phosphoproteomics identify arachidonic-acid-regulated signal transduction pathways modulating macrophage functions with implications for ovarian cancer.Theranostics2021;11:1377-95 PMCID:PMC7738879

[52] Song NJ,Kim KH.Butein is a novel anti-adipogenic compound.J Lipid Res2013;54:1385-96 PMCID:PMC3622332

[53] Song NJ,Rajbhandari P.Prdm4 induction by the small molecule butein promotes white adipose tissue browning.Nat Chem Biol2016;12:479-81 PMCID:PMC4912433

[54] Taouis M.Is resistin the master link between inflammation and inflammation-related chronic diseases?.Mol Cell Endocrinol2021;533:111341

[55] Qiu L,Yu L,Jia XJ.Novel oncogenic and chemoresistance-inducing functions of resistin in ovarian cancer cells require miRNAs-mediated induction of epithelial-to-mesenchymal transition.Sci Rep2018;8:12522 PMCID:PMC6104088

[56] Su CM,Chi MJ.Resistin facilitates VEGF-C-associated lymphangiogenesis by inhibiting miR-186 in human chondrosarcoma cells.Biochem Pharmacol2018;154:234-42

[57] Kumar D,Puan KJ.Resistin expression in human monocytes is controlled by two linked promoter SNPs mediating NFKB p50/p50 binding and C-methylation.Sci Rep2019;9:15245 PMCID:PMC6811637

[58] Cioce M,Goparaju C,Carbone M.Autocrine CSF-1R signaling drives mesothelioma chemoresistance via AKT activation.Cell Death Dis2014;5:e1167 PMCID:PMC5424113