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Megakaryoblast ve bazofil hücre tipine sahip Ph (+) KML hücre hatlarının Wnt ve Frizzled ilişkili gen ifadelerinin karşılaştırılması

Yıl 2021, Cilt: 46 Sayı: 1, 318 - 324, 31.03.2021
https://doi.org/10.17826/cumj.795833

Öz

Amaç: Bu çalışmada blastik fazda farklı hücre serilerinde tutulum gösteren iki farklı Ph (+) kronik miyeloid lösemi (KML) hücre hattında, Wnt sinyal yolağı ana oyuncuları olan Wnt ve Frizzled genlerinin ifade farklılıklarının incelenmesi amaçlanmıştır.
Gereç ve Yöntem: Megakaryoblast (MEG-01) ve bazofil (KU812) hücre tipine sahip Ph (+) KML hücre hatları kültüre edildikten sonra, RNA izolasyonları gerçekleştirilmiştir. RNA konsantrasyonları spektrofotometrede belirlendikten sonra, cDNA çevrimi yapılmıştır. PCR ile Wnt ve Frizzled genlerinin ifade düzeyleri analiz edilmiş, iki hücre hattının Wnt ve Frizzled ekspresyon verileri birbirleri ile karşılaştırılmıştır.
Bulgular: Yapılan analizler sonrasında MEG-01 hücrelerinin Fzd1-9 genlerini, KU812’nin ise Fzd3, Fzd4, Fzd5, Fzd6, Fzd7, Fzd9 genlerini ifade ettiği gözlenmiştir. MEG-01 hücreleri Wnt2b ve Wnt3 genlerini eksprese ederken, KU812 hücrelerinin ise sadece Wnt3’ü eksprese ettiği saptanmıştır.
Sonuç: KU812 ve MEG-01 hücre hatlarının, Wnt sinyal genlerinin ifade düzeyleri açısından birbirinden farklılık gösterdiği belirlenmiştir. İki KML Ph (+) hücre hattında gözlenen Wnt sinyal genlerinin ifade düzeyleri arasındaki farklara, hücrenin orjini, füzyon bölgeleri ve hücre tipine özgü olabilecek sinyal ileti uyarı ve yolaklarındaki farklılıkların neden olabileceği düşünülmektedir. Bu nedenle model hücre hatları kullanılarak yapılan çalışmalar da birden fazla hücre hattının seçilmesi önerilmektedir.

Destekleyen Kurum

Dokuz Eylül Üniversitesi, İKÇÜ

Kaynakça

  • Jung YS, Park J Il. Wnt signaling in cancer: therapeutic targeting of Wnt signaling beyond β-catenin and the destruction complex. Experimental and Molecular Medicine. 2020. PMID: 32037398
  • Zhong Z, Virshup DM. Wnt signaling and drug resistance in cancer. Mol Pharmacol. 2020; PMID: 31787618
  • Van Andel H, Kocemba KA, Spaargaren M, Pals ST. Aberrant Wnt signaling in multiple myeloma: molecular mechanisms and targeting options. Leukemia. 2019. PMID: 30770859
  • Flores-Hernández E, Velázquez DM, Castañeda-Patlán MC, Fuentes-García G, Fonseca-Camarillo G, Yamamoto-Furusho JK, Romero-Avila MT, García-Sáinz JA, Robles-Flores M. Canonical and non-canonical Wnt signaling are simultaneously activated by Wnts in colon cancer cells. Cell Signal. 2020; PMID: 32283254
  • Chiarini F, Paganelli F, Martelli AM, Evangelisti C. The Role Played by Wnt/β-Catenin Signaling Pathway in Acute Lymphoblastic Leukemia. Int J Mol Sci [Internet]. MDPI; 2020 Feb 7;21(3):1098. Available from: https://pubmed.ncbi.nlm.nih.gov/32046053
  • An X, Tiwari AK FAU - Sun Y, Sun Y FAU - Ding P-R, Ding PR FAU - Ashby CRJ, Ashby CR Jr FAU - Chen Z-S, ZS C. BCR-ABL tyrosine kinase inhibitors in the treatment of Philadelphia chromosome positive chronic myeloid leukemia: a review. PG - 1255-68 LID - 10.1016/j.leukres.2010.04.016 [doi]. Department of Pharmaceutical Sciences, College of Pharmacy and Allied Health Professions, St John’s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA.
  • Kleppe M, Levine RL. Targeting β-catenin in CML: Leukemia stem cells beware! Cell Stem Cell. 2012. Hu LH, Pu LF, Yang DD, Zhang C, Wang HP, Ding YY, Li MM, Zhai ZM, Xiong S. How to detect the rare BCR‑ABL (e14a3) transcript: A case report and literature review. Oncology Letters. 2017.
  • Balatzenko G, Vundinti BR, Margarita G. Correlation between the type of bcr-abl transcripts and blood cell counts in chronic myeloid leukemia - a possible influence of mdr1 gene expression. Hematol Rev. 2011.
  • Farhat-Maghribi S, Habbal W, Monem F. Frequency of BCR-ABL Transcript Types in Syrian CML Patients. J Oncol. 2016.
  • Sharma P, Kumar L FAU - Mohanty S, Mohanty S FAU - Kochupillai V, Kochupillai V. Response to Imatinib mesylate in chronic myeloid leukemia patients with variant BCR-ABL fusion transcripts. PG - 241-7 LID - 10.1007/s00277-009-0822-7 [doi]. Department of Medical Oncology, Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India. FAU - Kumar, Lalit; 2009.
  • Sercan HO, Pehlivan M, Simsek O, Ates H, Sercan Z. Induction of apoptosis increases expression of non-canonical WNT genes in myeloid leukemia cell lines. Oncol Rep. 2007; PMID: 17982645
  • www.atcc.org ATCC [Internet]. Available from:www.atcc.org.https://www.lgcstandardsatcc.org/Products/Cells_and_Microorganisms/Bacteria.aspx?geo_country=tr
  • Grassi S, Palumbo S, Mariotti V, Liberati D, Guerrini F, Ciabatti E, Salehzadeh S, Baratè C, Balducci S, Ricci F, Buda G, Iovino L, Mazziotta F, Ghio F, Ercolano G, Di Paolo A, Cecchettini A, Baldini C, Mattii L, Pellegrini S, Petrini M, Galimberti S. The WNT pathway is relevant for the BCR-ABL1-independent resistance in chronic myeloid leukemia. Front Oncol. 2019.
  • Hu J, Feng M, ZL L, Liu Y, ZL H, Li H, WL F. Potential role of Wnt/β-catenin signaling in blastic transformation of chronic myeloid leukemia: cross talk between β-catenin and BCR-ABL. Department of Clinical Hematology, Key Laboratory of Laboratory Medical Diagnostics Designated by the Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Sercan Z, Pehlivan M, Sercan HO. Expression profile of WNT, FZD and sFRP genes in human hematopoietic cells. Leuk Res. 2010; PMID: 20206998
  • Liu F, Wang B, Wang Q, Qi Z, Chen C, Kong LL, Chen JY, Liu X, Wang A, Hu C, Wang W, Wang H, Wu F, Ruan Y, Qi S, Liu J, Zou F, Hu Z, Wang W, Wang L, Zhang S, Yun CH, Zhai Z, Liu J, Liu Q. Discovery and characterization of a novel potent type II native and mutant BCR-ABL inhibitor (CHMFL-074) for Chronic Myeloid Leukemia (CML). Oncotarget. 2016; PMID: 27322145.
  • Peng Z, Luo HW, Yuan Y, Jing S, Huang SF, Li CL, Cao WX, Huang ZG, Feng WL. Growth of chronic myeloid leukemia cells is inhibited by infection with Ad-SH2-HA adenovirus that disrupts Grb2-Bcr-Abl complexes. Oncol Rep. 2011; PMID: 21369701.
  • Chu S, Li A, Singh H, Bhatia R. BCR-tyrosine 177 plays an essential role in Ras and Akt activation and in human hematopoietic progenitor transformation in chronic myelogenous leukemia. Cancer Res. 2007; PMID: 17638918.
  • Sekikawa T, Iwase S, Saito S, Arakawa Y, Agawa M, Horiguchi-Yamada J, Yamada H. JAS-R, a new megakaryo-erythroid leukemic cell line that secretes erythropoietin. Anticancer Res. 2006; PMID: 16619478.
  • Haaß W, Kleiner H, Weiß C, Haferlach C, Schlegelberger B, Müller MC, Hehlmann R, Hofmann WK, Fabarius A, Seifarth W. Clonal evolution and blast crisis correlate with enhanced proteolytic activity of Separase in BCR-ABL b3a2 fusion type CML under imatinib therapy. PLoS One. 2015; PMID: 26087013.
  • Gong Z, Medeiros LJ, Cortes JE, Zheng L, Khoury JD, Wang W, Tang G, Loghavi S, Luthra R, Yang W, Kantarjian HM, Hu S. Clinical and prognostic significance of e1a2 BCR-ABL1 transcript subtype in chronic myeloid leukemia. Blood Cancer Journal. 2017. PMID: 28708130.

Comparison of Wnt and Frizzled related gene expressions of Ph (+) CML cell lines with megakaryoblast and basophil cell types

Yıl 2021, Cilt: 46 Sayı: 1, 318 - 324, 31.03.2021
https://doi.org/10.17826/cumj.795833

Öz

Purpose: The aim of this study was to investigate the expression changes in Wnt and Frizzled genes that initiate Wnt signaling in two different Ph (+) chronic myeloid leukemia (CML) cell lines.
Materials and Methods: Total RNA isolations were performed from Ph (+) CML cell lines with megakaryoblast and basophil cell types. RNA concentrations was evaluated by spectrophotometry and reverted to cDNA. Expression levels of Wnt and Frizzled genes were analyzed by PCR and data of two cell lines were compared with each other.
Results: In the analyzes, it was observed that MEG-01 cells expressed Fzd1-9 genes, while KU812 expressed Fzd3, Fzd4, Fzd5, Fzd6, Fzd7, Fzd9 genes. It was determined that MEG-01 cells expressed Wnt2b and Wnt3 genes, while KU812 cells expressed only Wnt3 gene.
Conclusion: KU812, MEG-01 cell lines differ from each other in terms of expression levels of Wnt signaling genes. It is thought that the differences between the expression levels of Wnt genes observed in the two CML Ph (+) cell lines may be caused by the differences in the cell's origin, fusion regions and cell type-specific signal transduction stimuli and pathways. Therefore, it is recommended to select more than one cell line in studies using model cell lines.

Kaynakça

  • Jung YS, Park J Il. Wnt signaling in cancer: therapeutic targeting of Wnt signaling beyond β-catenin and the destruction complex. Experimental and Molecular Medicine. 2020. PMID: 32037398
  • Zhong Z, Virshup DM. Wnt signaling and drug resistance in cancer. Mol Pharmacol. 2020; PMID: 31787618
  • Van Andel H, Kocemba KA, Spaargaren M, Pals ST. Aberrant Wnt signaling in multiple myeloma: molecular mechanisms and targeting options. Leukemia. 2019. PMID: 30770859
  • Flores-Hernández E, Velázquez DM, Castañeda-Patlán MC, Fuentes-García G, Fonseca-Camarillo G, Yamamoto-Furusho JK, Romero-Avila MT, García-Sáinz JA, Robles-Flores M. Canonical and non-canonical Wnt signaling are simultaneously activated by Wnts in colon cancer cells. Cell Signal. 2020; PMID: 32283254
  • Chiarini F, Paganelli F, Martelli AM, Evangelisti C. The Role Played by Wnt/β-Catenin Signaling Pathway in Acute Lymphoblastic Leukemia. Int J Mol Sci [Internet]. MDPI; 2020 Feb 7;21(3):1098. Available from: https://pubmed.ncbi.nlm.nih.gov/32046053
  • An X, Tiwari AK FAU - Sun Y, Sun Y FAU - Ding P-R, Ding PR FAU - Ashby CRJ, Ashby CR Jr FAU - Chen Z-S, ZS C. BCR-ABL tyrosine kinase inhibitors in the treatment of Philadelphia chromosome positive chronic myeloid leukemia: a review. PG - 1255-68 LID - 10.1016/j.leukres.2010.04.016 [doi]. Department of Pharmaceutical Sciences, College of Pharmacy and Allied Health Professions, St John’s University, 8000 Utopia Parkway, Jamaica, NY 11439, USA.
  • Kleppe M, Levine RL. Targeting β-catenin in CML: Leukemia stem cells beware! Cell Stem Cell. 2012. Hu LH, Pu LF, Yang DD, Zhang C, Wang HP, Ding YY, Li MM, Zhai ZM, Xiong S. How to detect the rare BCR‑ABL (e14a3) transcript: A case report and literature review. Oncology Letters. 2017.
  • Balatzenko G, Vundinti BR, Margarita G. Correlation between the type of bcr-abl transcripts and blood cell counts in chronic myeloid leukemia - a possible influence of mdr1 gene expression. Hematol Rev. 2011.
  • Farhat-Maghribi S, Habbal W, Monem F. Frequency of BCR-ABL Transcript Types in Syrian CML Patients. J Oncol. 2016.
  • Sharma P, Kumar L FAU - Mohanty S, Mohanty S FAU - Kochupillai V, Kochupillai V. Response to Imatinib mesylate in chronic myeloid leukemia patients with variant BCR-ABL fusion transcripts. PG - 241-7 LID - 10.1007/s00277-009-0822-7 [doi]. Department of Medical Oncology, Institute Rotary Cancer Hospital, All India Institute of Medical Sciences, Ansari Nagar, New Delhi 110029, India. FAU - Kumar, Lalit; 2009.
  • Sercan HO, Pehlivan M, Simsek O, Ates H, Sercan Z. Induction of apoptosis increases expression of non-canonical WNT genes in myeloid leukemia cell lines. Oncol Rep. 2007; PMID: 17982645
  • www.atcc.org ATCC [Internet]. Available from:www.atcc.org.https://www.lgcstandardsatcc.org/Products/Cells_and_Microorganisms/Bacteria.aspx?geo_country=tr
  • Grassi S, Palumbo S, Mariotti V, Liberati D, Guerrini F, Ciabatti E, Salehzadeh S, Baratè C, Balducci S, Ricci F, Buda G, Iovino L, Mazziotta F, Ghio F, Ercolano G, Di Paolo A, Cecchettini A, Baldini C, Mattii L, Pellegrini S, Petrini M, Galimberti S. The WNT pathway is relevant for the BCR-ABL1-independent resistance in chronic myeloid leukemia. Front Oncol. 2019.
  • Hu J, Feng M, ZL L, Liu Y, ZL H, Li H, WL F. Potential role of Wnt/β-catenin signaling in blastic transformation of chronic myeloid leukemia: cross talk between β-catenin and BCR-ABL. Department of Clinical Hematology, Key Laboratory of Laboratory Medical Diagnostics Designated by the Ministry of Education, Chongqing Medical University, Chongqing, China.
  • Sercan Z, Pehlivan M, Sercan HO. Expression profile of WNT, FZD and sFRP genes in human hematopoietic cells. Leuk Res. 2010; PMID: 20206998
  • Liu F, Wang B, Wang Q, Qi Z, Chen C, Kong LL, Chen JY, Liu X, Wang A, Hu C, Wang W, Wang H, Wu F, Ruan Y, Qi S, Liu J, Zou F, Hu Z, Wang W, Wang L, Zhang S, Yun CH, Zhai Z, Liu J, Liu Q. Discovery and characterization of a novel potent type II native and mutant BCR-ABL inhibitor (CHMFL-074) for Chronic Myeloid Leukemia (CML). Oncotarget. 2016; PMID: 27322145.
  • Peng Z, Luo HW, Yuan Y, Jing S, Huang SF, Li CL, Cao WX, Huang ZG, Feng WL. Growth of chronic myeloid leukemia cells is inhibited by infection with Ad-SH2-HA adenovirus that disrupts Grb2-Bcr-Abl complexes. Oncol Rep. 2011; PMID: 21369701.
  • Chu S, Li A, Singh H, Bhatia R. BCR-tyrosine 177 plays an essential role in Ras and Akt activation and in human hematopoietic progenitor transformation in chronic myelogenous leukemia. Cancer Res. 2007; PMID: 17638918.
  • Sekikawa T, Iwase S, Saito S, Arakawa Y, Agawa M, Horiguchi-Yamada J, Yamada H. JAS-R, a new megakaryo-erythroid leukemic cell line that secretes erythropoietin. Anticancer Res. 2006; PMID: 16619478.
  • Haaß W, Kleiner H, Weiß C, Haferlach C, Schlegelberger B, Müller MC, Hehlmann R, Hofmann WK, Fabarius A, Seifarth W. Clonal evolution and blast crisis correlate with enhanced proteolytic activity of Separase in BCR-ABL b3a2 fusion type CML under imatinib therapy. PLoS One. 2015; PMID: 26087013.
  • Gong Z, Medeiros LJ, Cortes JE, Zheng L, Khoury JD, Wang W, Tang G, Loghavi S, Luthra R, Yang W, Kantarjian HM, Hu S. Clinical and prognostic significance of e1a2 BCR-ABL1 transcript subtype in chronic myeloid leukemia. Blood Cancer Journal. 2017. PMID: 28708130.
Toplam 21 adet kaynakça vardır.

Ayrıntılar

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

Melek Pehlivan 0000-0001-8755-4812

Hakkı Ogün Sercan 0000-0002-2449-1794

Yayımlanma Tarihi 31 Mart 2021
Kabul Tarihi 21 Ekim 2020
Yayımlandığı Sayı Yıl 2021 Cilt: 46 Sayı: 1

Kaynak Göster

MLA Pehlivan, Melek ve Hakkı Ogün Sercan. “Megakaryoblast Ve Bazofil hücre Tipine Sahip Ph (+) KML hücre hatlarının Wnt Ve Frizzled ilişkili Gen Ifadelerinin karşılaştırılması”. Cukurova Medical Journal, c. 46, sy. 1, 2021, ss. 318-24, doi:10.17826/cumj.795833.