Buket KOSOVA, Asu Fergün YILMAZ, Burçin KAYMAZ, Filiz VURAL, Fahri ŞAHİN, Çağdaş AKTAN, Güray SAYDAM, Melda CÖMERT, Nur SOYER, Ajda GÜNEŞ

Determining expression of miRNAs that potentially regulate STAT5A and 5B in dasatinib-sensitive K562 cells

In the era of tyrosine kinase inhibitors, resistance still constitutes a problem in chronic myeloid leukemia (CML) patients;thus, new pathway-specific inhibitors like miRNAs have become important in the treatment of refractory patients. There are nosatisfying data regarding the miRNAs and anti-miRNA treatment targeting STAT5A and 5B. In this study, we first researched the effectof dasatinib on apoptosis in the CML cell line K562. The expressions of miRNAs possibly targeting both STAT5A and 5B were thendetermined. The down- and upregulation of the miRNAs were compared using the ΔΔCT method. At the last stage of the study, we useda new primer probe in order to validate the results. The level of hsa-miR-940 was decreased 4.4 times and the levels of hsa-miR-527 andhsa-miR-518a-5p were increased 12.1 and 8 times, respectively, in the dasatinib-treated group when compared to the control group.We detected similar results in the validation step. As a conclusion, we determined the expression profiles of miRNAs targeting STAT5Aand 5B that had an important role in the pathogenesis of CML. The data obtained could lead to determining new therapeutic targetsfor CML patients.

PDF

___

Agirre X, Jiménez-Velasco A, San José-Enériz E, Garate L, Bandrés E, Cordeu L, Aparicio O, Saez B, Navarro G, Vilas-Zornoza A et al. (2008). Down-regulation of hsa-miR-10a in chronic myeloid leukemia CD34+ cells increases USF2-mediated cell growth. Mol Cancer Res 6: 1830-1840.
Apperley JF (2007). Part I: Mechanisms of resistance to imatinib in chronic myeloid leukaemia. Lancet Oncol 8: 1018-1029. Avery-Kiejda KA, Braye SG, Mathe A , Forbes JF , Scott RJ (2014). Decreased expression of key tumour suppressor microRNAs is associated with lymph node metastases in triple negative breast cancer. BMC Cancer 31: 14-51.
Bartel DP (2004). MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116: 281-297.
Benekli M, Baumann H, Wetzler M (2009). Targeting signal transducer and activator of transcription signaling pathway in leukemias. J Clin Oncol 27: 4422-4432.
Bowman T, Garcia R, Turkson J, Jove R (2000). STATs in oncogenesis. Oncogene 19: 2474-2488.
Bueno MJ, Pérez de Castro I, Gómez de Cedrón M, Santos J, Calin GA, Cigudosa JC, Croce CM, Fernández-Piqueras J, Malumbres M (2008). Genetic and epigenetic silencing of microRNA-203 enhances ABL1 and BCR-ABL1 oncogene expression. Cancer Cell 13: 496-506.
Calin GA, Croce CM (2006). MicroRNA signatures in human cancers. Nat Rev Cancer 6: 857-866.
Calin GA, Dumitru CD, Shimizu M, Bichi R, Zupo S, Noch E, Aldler H, Rattan S, Keating M, Rai K et al. (2002), Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. P Natl Acad Sci USA 99: 15524-15529.
Cheng AM, Byrom MW , Shelton J , Ford LP (2005). Antisense inhibition of human miRNAs and indications for an involvement of miRNA in cell growth and apoptosis. Nucleic Acids Res 33: 1290-1297.
Dalgiç CT, Kaymaz BT, Dalmizrak A, Cömert Özkan M, Kosova B, Şahin F, Saydam G (2015). Investigating the role of JAK/STAT pathway on dasatinib-induced apoptosis for CML cell model K562. Clin Lymphoma Myeloma Leuk 15: 161-166.
Dixon-McIver A, East P, Mein CA, Cazier JB, Molloy G, Chaplin T, Lister TA, Young BD, Debernardi S (2008). Distinctive patterns of microRNA expression associated with karyotype in acute myeloid leukaemia. PLoS One 3: e2141.
Eiring AM, Harb JG, Neviani P, Garton C, Oaks JJ, Spizzo R, Liu S, Schwind S, Santhanam R, Hickey CJ et al. (2010). miR- 328 functions as an RNA decoy to modulate hnRNP E2 regulation of mRNA translation in leukemic blasts. Cell 140: 652-665.
Farazi TA, Spitzer JI, Morozov P, Tuschl T (2011). miRNAs in human cancer. J Pathol 223: 102-115.
Ferreira AF, de Oliveira GL, Tognon R, Collassanti MD, Zanichelli MA, Hamerschlak N, de Souza AM, Covas DT, Kashima S, de C a s t ro FA (2015). Apoptosis-related gene expression profile in chronic myeloid leukemia patients after imatinib mesylate and dasatinib therapy. Acta Haematol 133: 354-364.
Flamant S, Ritchie W, Guilhot J, Holst J, Bonnet ML, Chomel JC, Guilhot F, Turhan AG, Rasko JE (2010). Micro-RNA response to imatinib mesylate in patients with chronic myeloid leukemia. Haematologica 95: 1325-1333.
Garzon R, Garofalo M, Martelli MP, Briesewitz R, Wang L, Fernandez- Cymering C, Volinia S, Liu CG, Schnittger S, Haferlach T et al. (2008a). Distinctive microRNA signature of acute myeloid leukemia bearing cytoplasmic mutated nucleophosmin. P Natl Acad Sci USA 105: 3945-3950.
Garzon R, Pichiorri F, Palumbo T, Visentini M, Aqeilan R, Cimmino A, Wang H, Sun H, Volinia S, Alder H et al. (2007). MicroRNA gene expression during retinoic acid-induced differentiation of human acute promyelocytic leukemia. Oncogene 26: 4148- 4157.
Garzon R, Volinia S, Liu CG, Fernandez-Cymering C, Palumbo T, Pichiorri F, Fabbri M, Coombes K, Alder H, Nakamura T (2008b). MicroRNA signatures associated with cytogenetics and prognosis in acute myeloid leukemia. Blood 111: 3183- 3189.
Groffen J, Stephenson JR, Heisterkamp N, Groffen J, Stephenson JR, Heisterkamp N, de Klein A, Bartram CR, Grosveld G (1984). Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22. Cell 36: 93-99.
Han YC, Park CY, Bhagat G, Zhang J, Wang Y, Fan JB, Liu M, Zou Y, Weissman IL, Gu H (2010). microRNA-29a induces aberrant self-renewal capacity in hematopoietic progenitors, biased myeloid development, and acute myeloid leukemia. J Exp Med 207: 475-489.
Heo SK, Noh EK, Yoon DJ, Jo JC, Park JH, Kim H (2014). Dasatinib accelerates valproic acid-induced acute myeloid leukemia cell death by regulation of differentiation capacity. PLoS One 9: e98859.
Hershkovitz-Rokah O, Modai S, Pasmanik-Chor M, Toren A, Shomron N, Raanani P, Shpilberg O, Granot G (2015). Restoration of miR-424 suppresses BCR-ABL activity and sensitizes CM cells to imatinib treatment. Cancer Lett 360: 245-256.
Kantarjian H, Sawyers C, Hochhaus A, Guilhot F, Schiffer C, Gambacorti-Passerini C, Niederwieser D, Resta D, Capdeville R, Zoellner U et al. (2002). Hematologic and cytogenetic responses to imatinib mesylate in chronic myelogenous leukemia. N Engl J Med. 346: 645-652.
Kaymaz BT, Çetintaş VB, Aktan Ç, Kosova B (2014). MicroRNA- 520a-5p displays a therapeutic effect upon chronic myelogenous leukemia cells by targeting STAT3 and enhances the anticarcinogenic role of capsaicin. Tumour Biol 35: 8733- 8742.
Kaymaz BT, Günel NS, Ceyhan M, Çetintaş VB, Ö zel B, Yandım MK, Kıpçak S, Aktan Ç, Gökbulut AA, Baran Y et al. (2015). Revealing genome-wide mRNA and microRNA expression patterns in leukemic cells highlighted hsa-miR-2278 as a tumor suppressor for regain of chemotherapeutic imatinib response due to targeting STAT5A. Tumour Biol 36: 7915- 7927.
Kosova B, Tezcanli B, Ekiz HA, Cakir Z, Selvi N, Dalmizrak A, Kartal M, Gunduz U, Baran Y (2010). Suppression of STAT5A increases chemotherapeutic sensitivity in imatinib-resistant and imatinib-sensitive K562 cells. Leuk Lymphoma 51: 1895- 1901.
Li Y, Wang H, Tao K, Xiao Q, Huang Z, Zhong L, Cao W, Wen J, Feng W (2013). miR-29b suppresses CML cell proliferation and induces apoptosis via regulation of BCR/ABL1 protein. Exp Cell Res 319: 109-101.
Liu L, Wang S, Chen R, Wu Y, Zhang B, Huang S, Zhang J, Xiao F, Wang M, Liang Y (2012). Myc induced miR-144/451 contributes to the acquired imatinib resistance in chronic myelogenous leukemia cell K562. Biochem Biophys Res Commun 425: 368-373.
Liu X, Ge X, Zhang Z, Zhang X, Chang J, Wu Z, Tang W, Gan L, Sun M, Li J (2015). MicroRNA-940 promotes tumor cell invasion and metastasis by downregulating ZNF24 in gastric cancer. Oncotarget 6: 25418-25428.
Mi S, Lu J, Sun M, Li Z, Zhang H, Neilly MB, Wang Y, Qian Z, Jin J, Zhang Y et al. (2007). MicroRNA expression signatures accurately discriminate acute lymphoblastic leukemia from acute myeloid leukemia. P Natl Acad Sci USA 104: 19971- 19976.
Miao L, Xiong X, Lin Y, Cheng Y, Lu J, Zhang J, Cheng N (2014). miR- 203 inhibits tumor cell migration and invasion via caveolin-1 in pancreatic cancer cells. Oncol Lett 7: 658-662.
Port M, Glaesener S, Ruf C, Riecke A, Bokemeyer C, Meineke V, Honecker F, Abend M (2011). Micro-RNA expression in cisplatin resistant germ cell tumor cell lines. Mol Cancer 10: 52.
Ramkissoon SH, Mainwaring LA, Ogasawara Y, Keyvanfar K, McCoy JP Jr, Sloand EM, Kajigaya S, Young NS (2006). Hematopoietic- specific microRNA expression in human cells. Leuk Res 30: 643-647.
Rawlings JS, Rosler KM, Harrison DA (2004). The JAK/STAT signaling pathway. J Cell Sci 117: 1281-1283.
Rodriguez Calleja L, Jacques C, Lamoureux F, Baudhuin M, Tellez Gabriel M, Quillard T, Sahay D, Perrot P, Amiaud J, Charrier C et al. (2016). ΔNp63α silences a microRNA program to aberrantly initiate a wound healing program that promotes TGFβ-induced metastasis. Cancer Res 76: 3236-3251.
Rokah OH, Granot G, Ovcharenko A, Modai S, Pasmanik-Chor M, Toren A, Shomron N, Shpilberg O (2012). Downregulation of miR-31, miR-155 and miR-564 in chronic myeloid leukemia cells. PLoS One 7: 35501.
Rubie C, Kruse B, Frick VO, Kölsch K, Ghadjar P, Wagner M, Grässer F, Wagenpfeil S, Glanemann M (2014). Chemokine receptor CCR6 expression is regulated by miR-518a-5p in colorectal cancer cells. J Transl Med 12: 48.
Saumet A, Vetter G, Bouttier M, Portales-Casamar E, Wasserman WW, Maurin T, Mari B, Barbry P, Vallar L, Friederich E et al. (2009). Transcriptional repression of microRNA genes by PML-RARA increases expression of key cancer proteins in acute promyelocytic leukemia. Blood 113: 412-421.
Shi Y, Gao X, Hu Q, Li X, Xu J, Lu S, Liu Y, Xu C, Jiang D, Lin J et al. (2016). PIK3C2A is a gene-specific target of microRNA- 518a-5p in imatinib mesylate-resistant gastrointestinal stromal tumor. Lab Invest 96: 652-660.
Simara P, Stejskal S, Koutna I, Potesil D, Tesarova L, Potesilova M, Zdrahal Z, Mayer J (2013). Apoptosis in chronic myeloid leukemia cells transiently treated with imatinib or dasatinib is caused by residual BCR ABL kinase inhibition. Am J Hematol 88: 385-393.
Song B, Zhang C, Li G, Jin G, Liu C (2015). MiR-940 inhibited pancreatic ductal adenocarcinoma growth by targeting MyD88. Cell Physiol Biochem 35: 1167-1177.
Spizzo R, Nicoloso MS, Croce CM, Calin GA (2009). SnapShot: MicroRNAs in cancer. Cell 137: 586-586.
Steelman LS, Pohnert SC, Shelton JG, Franklin RA, Bertrand FE, McCubrey JA (2004). JAK/STAT, Raf/MEK/ERK, PI3K/Akt and BCR-ABL in cell cycle progression and leukemogenesis. Leukemia 18: 189-218.
Suresh S, McCallum L, Lu W, Lazar N, Perbal B, Irvine AE (2011). MicroRNAs 130a/b are regulated by BCR-ABL and downregulate expression of CCN3 in CML. J Cell Commun Signal 5: 183-191.
Troppan K, Wenzl K, Deutsch A, Ling H, Neumeister P, Pichler M (2014). MicroRNAs in diffuse large b-cell lymphoma: implications for pathogenesis, diagnosis, prognosis and therapy. Anticancer Res 34: 557-564.
Venturini L, Battmer K, Castoldi M, Schultheis B, Hochhaus A, Muckenthaler MU, Ganser A, Eder M, Scherr M (2007). Expression of the miR-17-92 polycistron in chronic myeloid leukemia (CML) CD34+ cells. Blood 109: 4399-4405.
Xu C, Fu H, Gao L, Wang L, Wang W, Li J, Li Y, Dou L, Gao X, Luo X et al. (2014). BCR-ABL/GATA1/miR-138 mini circuitry contributes to the leukemogenesis of chronic myeloid leukemia. Oncogene 33: 44-54.
Yu H, Hove R (2004). The stats of cancer: new molecular targets come of age. Nat Rev Cancer 4: 97-105.
Yuan B, Liang Y, Wang D, Luo F (2015). MiR-940 inhibits hepatocellular carcinoma growth and correlates with prognosis of hepatocellular carcinoma patients. Cancer Sci 106: 819-824.
Zheng G, Xiong Y, Xu W, Wang Y, Chen F, Wang Z, Yan Z (2014). A two-microRNA signature as a potential biomarker for early gastric cancer. Oncol Lett 7: 679-684.
Zimmerman EI, Dollins CM, Crawford M, Grant S, Nana-Sinkam SP, Richards KL, Hammond SM, Graves LM (2010). Lyn kinase-dependent regulation of miR181 and myeloid cell leukemia-1 expression: implications for drug resistance in myelogenous leukemia. Mol Pharmacol 78: 811-817.