Araştırma Makalesi
BibTex RIS Kaynak Göster

Global DNA Methylation analysis of imatinib resistant and sensitive K562 cells

Yıl 2024, Cilt: 7 Sayı: 1, 13 - 17, 14.05.2024
https://doi.org/10.33713/egetbd.1450605

Öz

OBJECTIVE: Chronic myeloid leukemia (CML) is a hematological disease which is known by the presence of Philadelphia chromosome (Ph+). BCR-ABL protein is expressed by Ph+ chromosome, represents constant increased tyrosine kinase activity. Imatinib (IMA) is a tyrosine kinase inhibitor (TKI) which is utilized as a first line treatment in CML. Emergence of IMA resistance at some point of therapy leads to treatment failure. DNA methylation is considered to be the most stable epigenetic change and several studies have shown that epigenetic changes may play a role in drug resistance. We investigated the global methylation profile of IMA-sensitive K562S, IMA-resistant K562R and IMA-resistant and adherent K562R (K562R-adh) cells to determine whether epigenetic reprogramming is involved in the resistance to IMA and the change in phenotype due to this resistance.
MATERIAL AND METHODS: In this study, morphologically distinct, IMA-sensitive K562S and 5µM IMA-resistant K562R and K562R-adh in-vitro CML cell models were used to analyze the global DNA methylation profile. After DNA was isolated from the cells, global 5mC DNA methylation profiles were investigated by ELISA using equal amounts of DNA.
RESULTS: Compared to K562S, the global methylation of K562R showed an increase in DNA methylation profile, but this increase in methylation was not statistically significant. Whereas, a slight hypermethylation was observed in the DNA of the K562R-adh vs K562S and K562R-adh vs K562R which is statistically significant. We observed slight hypermethylation in IMA-resistant cells lines versus to the IMA-sensitive cell line.
CONCLUSION: Our observed differences in 5methyl-Cytosine on CpG islands (5mC) in K562S versus K562R and K562R-adh cell lines suggest that the DNA methylation alteration in resistant cells may partly contributed in phenotype switching.

Kaynakça

  • Groffen J, Stephenson JR, Heisterkamp N, de Klein A, Bartram CR, Grosveld G. Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22. Cell. 1984;36(1):93-9.
  • Ansari S, Verma M. Control of Ph(+) and additional chromosomal abnormalities in chronic myeloid leukemia by tyrosine kinase inhibitors. Med Oncol. 2023;40(8):237.
  • Bugler J, Kinstrie R, Scott MT, Vetrie D. Epigenetic Reprogramming and Emerging Epigenetic Therapies in CML. Front Cell Dev Biol. 2019;7:136.
  • Loscocco F, Visani G, Galimberti S, Curti A, Isidori A. BCR-ABL Independent Mechanisms of Resistance in Chronic Myeloid Leukemia. Front Oncol. 2019;9:939.
  • You R-I, Ho C-L, Hung H-M, Hsieh Y-F, Ju J-C, Chao T-Y. Identification of DNA methylation biomarkers in imatinib resistant chronic myeloid leukemia cells. Genomic Medicine, Biomarkers, and Health Sciences. 2012;4(1-2):12-5.
  • Hekmatshoar Y, Ozkan T, Altinok Gunes B, Bozkurt S, Karadag A, Karabay AZ, et al. Characterization of imatinib-resistant K562 cell line displaying resistance mechanisms. Cell Mol Biol (Noisy-le-grand). 2018;64(6):23-30.
  • Kaehler M, Litterst M, Kolarova J, Bohm R, Bruckmueller H, Ammerpohl O, et al. Genome‑wide expression and methylation analyses reveal aberrant cell adhesion signaling in tyrosine kinase inhibitor‑resistant CML cells. Oncol Rep. 2022;48(2).
  • Kemper K, de Goeje PL, Peeper DS, van Amerongen R. Phenotype switching: tumor cell plasticity as a resistance mechanism and target for therapy. Cancer Res. 2014;74(21):5937-41.
  • Urbanova M, Buocikova V, Trnkova L, Strapcova S, Kajabova VH, Melian EB, et al. DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines. Int J Mol Sci. 2022;23(4).
  • Koschmieder S, Vetrie D. Epigenetic dysregulation in chronic myeloid leukaemia: A myriad of mechanisms and therapeutic options. Semin Cancer Biol. 2018;51:180-97.
  • Leo E. DNA Methylation in Chronic Myeloid Leukemia. Journal of Molecular and Genetic Medicine. 2016;10(02).
  • Deaton AM, Bird A. CpG islands and the regulation of transcription. Genes Dev. 2011;25(10):1010-22.
  • Byun HM, Eshaghian S, Douer D, Trent J, Garcia-Manero G, Bhatia R, et al. Impact of Chromosomal Rearrangement upon DNA Methylation Patterns in Leukemia. Open Med (Wars). 2017;12:76-85.
  • Jiang D, Hong Q, Shen Y, Xu Y, Zhu H, Li Y, et al. The diagnostic value of DNA methylation in leukemia: a systematic review and meta-analysis. PLoS One. 2014;9(5):e96822.
  • Hekmatshoar Y, Karadag Gurel A, Ozkan T, Rahbar Saadat Y, Koc A, Karabay AZ, et al. Phenotypic and functional characterization of subpopulation of Imatinib resistant chronic myeloid leukemia cell line. Adv Med Sci. 2023;68(2):238-48.
  • Bhamidipati PK, Kantarjian H, Cortes J, Cornelison AM, Jabbour E. Management of imatinib-resistant patients with chronic myeloid leukemia. Ther Adv Hematol. 2013;4(2):103-17.
  • Lebecque B, Bourgne C, Vidal V, Berger MG. DNA Methylation and Intra-Clonal Heterogeneity: The Chronic Myeloid Leukemia Model. Cancers (Basel). 2021;13(14).
  • Galle E, Thienpont B, Cappuyns S, Venken T, Busschaert P, Van Haele M, et al. DNA methylation-driven EMT is a common mechanism of resistance to various therapeutic agents in cancer. Clin Epigenetics. 2020;12(1):27.
  • Tam WL, Weinberg RA. The epigenetics of epithelial-mesenchymal plasticity in cancer. Nat Med. 2013;19(11):1438-49.
  • Ko TK, Javed A, Lee KL, Pathiraja TN, Liu X, Malik S, et al. An integrative model of pathway convergence in genetically heterogeneous blast crisis chronic myeloid leukemia. Blood. 2020;135(26):2337-53.
  • Guru SA, Sumi MP, Mir R, Beg MMA, Koner BC, Saxena A. Aberrant hydroxymethylation in promoter CpG regions of genes related to the cell cycle and apoptosis characterizes advanced chronic myeloid leukemia disease, poor imatinib respondents and poor survival. BMC Cancer. 2022;22(1):405.
  • Baykal-Kose S, Acikgoz E, Yavuz AS, Gonul Geyik O, Ates H, Sezerman OU, et al. Adaptive phenotypic modulations lead to therapy resistance in chronic myeloid leukemia cells. PLoS One. 2020;15(2):e0229104.

İmatinibe dirençli ve duyarlı K562 hücrelerinin global DNA Metilasyon analizi

Yıl 2024, Cilt: 7 Sayı: 1, 13 - 17, 14.05.2024
https://doi.org/10.33713/egetbd.1450605

Öz

Kronik miyeloid lösemi (KML), Philadelphia kromozomunun (Ph+) varlığı ile bilinen hematolojik bir hastalıktır. BCR-ABL proteini Ph+ kromozomu tarafından ifade edilir ve sürekli olarak artmış tirozin kinaz aktivitesi göstermektedir. İmatinib (IMA), KML'de ilk basamak tedavi olarak kullanılan bir tirozin kinaz inhibitörüdür (TKI). Tedavinin bir noktasında IMA direncinin ortaya çıkması tedavi başarısızlığına yol açar. DNA metilasyonu en stabil epigenetik değişiklik olarak kabul edilir ve epigenetik değişikliklerin ilaç direncinde rol oynayabileceği çeşitli çalışmalarda gösterilmiştir. Bu çalışmada, IMA’ya karşı gelişen dirençte ve bu direnç nedeniyle fenotipte görülen değişimde epigenetik yeniden programlamanın rol alıp almadığını belirlemek amacıyla, IMA'ya duyarlı K562S, IMA'ya dirençli K562R ve IMA'ya dirençli ve yapışan K562R (K562R-adh) hücrelerinin global metilasyon profilini araştırdık.
GEREÇ VE YÖNTEM: Bu çalışmada, global DNA metilasyon profilini analiz etmek için morfolojik olarak farklılık gösteren, IMA'ya karşı duyarlı K562S ve 5µM IMA'ya karşı dirençli K562R ve K562R-adh in-vitro KML hücre modelleri kullanılmıştır. Hücrelerden DNA izole edildikten sonra, eşit miktardaki DNA’lar kullanılarak ELISA yöntemi ile global 5mC DNA metilasyon profilleri araştırılmıştır.
BULGULAR: K562R’nin global metilasyonu K562S'ye göre kıyaslandığında, DNA metilasyon profilinde artış saptanırken, metilasyondaki bu artış istatistiksel olarak anlamlı bulunmamıştır. Buna karşılık, K562R-adh ile K562S ve K562R-adh ile K562R kıyaslandığında, K562R-adh hücrelerinin global DNA metilasyon profilinde K562S ve K562R hücrelerinine göre istatistiksel olarak anlamlı derecede artış saptanmıştır.
SONUÇ: K562S, K562R ve K562R-adh hücre hatlarında CpG adalarındaki 5metil-Sitozin (5mC)'de gözlemlediğimiz farklılıklar, dirençli hücrelerdeki DNA metilasyon değişikliğinin fenotip değişimine kısmen katkıda bulunabileceğini düşündürmektedir.

Kaynakça

  • Groffen J, Stephenson JR, Heisterkamp N, de Klein A, Bartram CR, Grosveld G. Philadelphia chromosomal breakpoints are clustered within a limited region, bcr, on chromosome 22. Cell. 1984;36(1):93-9.
  • Ansari S, Verma M. Control of Ph(+) and additional chromosomal abnormalities in chronic myeloid leukemia by tyrosine kinase inhibitors. Med Oncol. 2023;40(8):237.
  • Bugler J, Kinstrie R, Scott MT, Vetrie D. Epigenetic Reprogramming and Emerging Epigenetic Therapies in CML. Front Cell Dev Biol. 2019;7:136.
  • Loscocco F, Visani G, Galimberti S, Curti A, Isidori A. BCR-ABL Independent Mechanisms of Resistance in Chronic Myeloid Leukemia. Front Oncol. 2019;9:939.
  • You R-I, Ho C-L, Hung H-M, Hsieh Y-F, Ju J-C, Chao T-Y. Identification of DNA methylation biomarkers in imatinib resistant chronic myeloid leukemia cells. Genomic Medicine, Biomarkers, and Health Sciences. 2012;4(1-2):12-5.
  • Hekmatshoar Y, Ozkan T, Altinok Gunes B, Bozkurt S, Karadag A, Karabay AZ, et al. Characterization of imatinib-resistant K562 cell line displaying resistance mechanisms. Cell Mol Biol (Noisy-le-grand). 2018;64(6):23-30.
  • Kaehler M, Litterst M, Kolarova J, Bohm R, Bruckmueller H, Ammerpohl O, et al. Genome‑wide expression and methylation analyses reveal aberrant cell adhesion signaling in tyrosine kinase inhibitor‑resistant CML cells. Oncol Rep. 2022;48(2).
  • Kemper K, de Goeje PL, Peeper DS, van Amerongen R. Phenotype switching: tumor cell plasticity as a resistance mechanism and target for therapy. Cancer Res. 2014;74(21):5937-41.
  • Urbanova M, Buocikova V, Trnkova L, Strapcova S, Kajabova VH, Melian EB, et al. DNA Methylation Mediates EMT Gene Expression in Human Pancreatic Ductal Adenocarcinoma Cell Lines. Int J Mol Sci. 2022;23(4).
  • Koschmieder S, Vetrie D. Epigenetic dysregulation in chronic myeloid leukaemia: A myriad of mechanisms and therapeutic options. Semin Cancer Biol. 2018;51:180-97.
  • Leo E. DNA Methylation in Chronic Myeloid Leukemia. Journal of Molecular and Genetic Medicine. 2016;10(02).
  • Deaton AM, Bird A. CpG islands and the regulation of transcription. Genes Dev. 2011;25(10):1010-22.
  • Byun HM, Eshaghian S, Douer D, Trent J, Garcia-Manero G, Bhatia R, et al. Impact of Chromosomal Rearrangement upon DNA Methylation Patterns in Leukemia. Open Med (Wars). 2017;12:76-85.
  • Jiang D, Hong Q, Shen Y, Xu Y, Zhu H, Li Y, et al. The diagnostic value of DNA methylation in leukemia: a systematic review and meta-analysis. PLoS One. 2014;9(5):e96822.
  • Hekmatshoar Y, Karadag Gurel A, Ozkan T, Rahbar Saadat Y, Koc A, Karabay AZ, et al. Phenotypic and functional characterization of subpopulation of Imatinib resistant chronic myeloid leukemia cell line. Adv Med Sci. 2023;68(2):238-48.
  • Bhamidipati PK, Kantarjian H, Cortes J, Cornelison AM, Jabbour E. Management of imatinib-resistant patients with chronic myeloid leukemia. Ther Adv Hematol. 2013;4(2):103-17.
  • Lebecque B, Bourgne C, Vidal V, Berger MG. DNA Methylation and Intra-Clonal Heterogeneity: The Chronic Myeloid Leukemia Model. Cancers (Basel). 2021;13(14).
  • Galle E, Thienpont B, Cappuyns S, Venken T, Busschaert P, Van Haele M, et al. DNA methylation-driven EMT is a common mechanism of resistance to various therapeutic agents in cancer. Clin Epigenetics. 2020;12(1):27.
  • Tam WL, Weinberg RA. The epigenetics of epithelial-mesenchymal plasticity in cancer. Nat Med. 2013;19(11):1438-49.
  • Ko TK, Javed A, Lee KL, Pathiraja TN, Liu X, Malik S, et al. An integrative model of pathway convergence in genetically heterogeneous blast crisis chronic myeloid leukemia. Blood. 2020;135(26):2337-53.
  • Guru SA, Sumi MP, Mir R, Beg MMA, Koner BC, Saxena A. Aberrant hydroxymethylation in promoter CpG regions of genes related to the cell cycle and apoptosis characterizes advanced chronic myeloid leukemia disease, poor imatinib respondents and poor survival. BMC Cancer. 2022;22(1):405.
  • Baykal-Kose S, Acikgoz E, Yavuz AS, Gonul Geyik O, Ates H, Sezerman OU, et al. Adaptive phenotypic modulations lead to therapy resistance in chronic myeloid leukemia cells. PLoS One. 2020;15(2):e0229104.
Toplam 22 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Biyokimya ve Hücre Biyolojisi (Diğer)
Bölüm Orijinal Araştırma
Yazarlar

Yalda Hekmatshoar 0000-0003-4683-074X

Erken Görünüm Tarihi 14 Mayıs 2024
Yayımlanma Tarihi 14 Mayıs 2024
Gönderilme Tarihi 11 Mart 2024
Kabul Tarihi 24 Nisan 2024
Yayımlandığı Sayı Yıl 2024 Cilt: 7 Sayı: 1

Kaynak Göster

EndNote Hekmatshoar Y (01 Mayıs 2024) Global DNA Methylation analysis of imatinib resistant and sensitive K562 cells. Ege Tıp Bilimleri Dergisi 7 1 13–17.

Creative Commons Lisansı


Bu eser Creative Commons Atıf-GayriTicari 4.0 Uluslararası Lisansı ile lisanslanmıştır.


13425                13428            13426            13433            13427