Differential expression patterns of leukaemia associated genes in leukaemia cell lines compared to healthy controls

Ang Pei-Shen, Rajesh Ramasamy, Noor Hamidah Hussin, Cheong Soon-Keng, Seow Heng-Fong, Maha Abdullah

Research output: Contribution to journalArticle

Abstract

Introduction: The phenotype and genotype of cancer cells portray hallmarks of cancer which may have clinical value. Cancer cell lines are ideal models to study and confrm these characteristics. We previously established two subtracted cDNA libraries with differentially expressed genes from an acute myeloid leukaemia patient with poor prognosis (PP) and good prognosis (GP). Objective: To compare gene expression of the leukaemia associated genes with selected biological characteristics in leukaemia cell lines and normal controls. Methodology: Expression of 28 PP genes associated with early fetal/embryonic development, HOX-related genes, hematopoiesis and aerobic glycolysis/hypoxia genes and 36 GP genes involved in oxidative phosphorylation, protein synthesis, chromatin remodelling and cell motility were examined in B-lymphoid (BV173, Reh and RS4;11) and myeloid (HL-60, K562) leukaemia cell lines after 72h in culture as well as peripheral blood mononuclear cells from healthy controls (N=5) using semi-quantitative polymerase chain reaction (PCR) method. Cell cycle profles were analysed on?ow cytometry while MTT cytotoxicity assay was used to determine drug resistance to epirubicin. Results: Genes expressed signifcantly higher in B-lymphoid leukaemia cell lines compared to healthy controls were mostly of the GP library i.e. oxidative phosphorylation (3/10), protein synthesis (4/11), chromatin remodelling (3/3) and actin cytoskeleton genes (1/5). Only two genes with signifcant difference were from the PP library. Cancer associated genes, HSPA9 and PSPH (GP library) and BCAP31 (PP library) were signifcantly higher in the B-lymphoid leukemia cell lines. No signifcant difference was observed between myeloid cell lines and healthy controls. This may also be due heterogeneity of cell lines studied. PBMC from healthy controls were not in cell cycle. G2/M profles and growth curves showed B-lymphoid cells just reaching plateau after 72 hour culture while myeloid cells were declining. IC50 values from cytotoxicity assay revealed myeloid cell lines had an average 13-fold higher drug resistance to epirubicin compared to B-lymphoid cell lines. Only CCL1, was expressed at least two-fold higher in myeloid compared to B-lymphoid cell lines. In contrast, MTRNR2, EEF1A1, PTMA, HLA-DR, C6orf115, PBX3, ENPP4, SELL, and IL3Ra were expressed more than 2-fold higher in B-lymphoid compared to myeloid cell lines studied here. Conclusion: Thus, B-lymphoid leukaemia cell lines here exhibited active, proliferating characteristics closer to GP genes. Higher expression of several genes in B-lymphoid compared to myeloid leukaemia cell lines may be useful markers to study biological differences including drug resistance between lineages.

Original languageEnglish
Pages (from-to)33-45
Number of pages13
JournalMalaysian Journal of Medicine and Health Sciences
Volume12
Issue number1
Publication statusPublished - 1 Jan 2016

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Leukemia
Cell Line
Genes
Myeloid Cells
Lymphocytes
Lymphoid Leukemia
Libraries
Drug Resistance
Epirubicin
Chromatin Assembly and Disassembly
Oxidative Phosphorylation
Cell Cycle
Embryonic and Fetal Development
Gene Expression
Neoplasms
Myeloid Leukemia
K562 Cells
Neoplasm Genes
Hematopoiesis
HLA-DR Antigens

Keywords

  • Chromatin remodelling genes
  • Gene profling
  • Leukaemia cell lines
  • Protein synthesis associated genes

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Differential expression patterns of leukaemia associated genes in leukaemia cell lines compared to healthy controls. / Pei-Shen, Ang; Ramasamy, Rajesh; Hussin, Noor Hamidah; Soon-Keng, Cheong; Heng-Fong, Seow; Abdullah, Maha.

In: Malaysian Journal of Medicine and Health Sciences, Vol. 12, No. 1, 01.01.2016, p. 33-45.

Research output: Contribution to journalArticle

Pei-Shen, Ang ; Ramasamy, Rajesh ; Hussin, Noor Hamidah ; Soon-Keng, Cheong ; Heng-Fong, Seow ; Abdullah, Maha. / Differential expression patterns of leukaemia associated genes in leukaemia cell lines compared to healthy controls. In: Malaysian Journal of Medicine and Health Sciences. 2016 ; Vol. 12, No. 1. pp. 33-45.
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AU - Pei-Shen, Ang

AU - Ramasamy, Rajesh

AU - Hussin, Noor Hamidah

AU - Soon-Keng, Cheong

AU - Heng-Fong, Seow

AU - Abdullah, Maha

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N2 - Introduction: The phenotype and genotype of cancer cells portray hallmarks of cancer which may have clinical value. Cancer cell lines are ideal models to study and confrm these characteristics. We previously established two subtracted cDNA libraries with differentially expressed genes from an acute myeloid leukaemia patient with poor prognosis (PP) and good prognosis (GP). Objective: To compare gene expression of the leukaemia associated genes with selected biological characteristics in leukaemia cell lines and normal controls. Methodology: Expression of 28 PP genes associated with early fetal/embryonic development, HOX-related genes, hematopoiesis and aerobic glycolysis/hypoxia genes and 36 GP genes involved in oxidative phosphorylation, protein synthesis, chromatin remodelling and cell motility were examined in B-lymphoid (BV173, Reh and RS4;11) and myeloid (HL-60, K562) leukaemia cell lines after 72h in culture as well as peripheral blood mononuclear cells from healthy controls (N=5) using semi-quantitative polymerase chain reaction (PCR) method. Cell cycle profles were analysed on?ow cytometry while MTT cytotoxicity assay was used to determine drug resistance to epirubicin. Results: Genes expressed signifcantly higher in B-lymphoid leukaemia cell lines compared to healthy controls were mostly of the GP library i.e. oxidative phosphorylation (3/10), protein synthesis (4/11), chromatin remodelling (3/3) and actin cytoskeleton genes (1/5). Only two genes with signifcant difference were from the PP library. Cancer associated genes, HSPA9 and PSPH (GP library) and BCAP31 (PP library) were signifcantly higher in the B-lymphoid leukemia cell lines. No signifcant difference was observed between myeloid cell lines and healthy controls. This may also be due heterogeneity of cell lines studied. PBMC from healthy controls were not in cell cycle. G2/M profles and growth curves showed B-lymphoid cells just reaching plateau after 72 hour culture while myeloid cells were declining. IC50 values from cytotoxicity assay revealed myeloid cell lines had an average 13-fold higher drug resistance to epirubicin compared to B-lymphoid cell lines. Only CCL1, was expressed at least two-fold higher in myeloid compared to B-lymphoid cell lines. In contrast, MTRNR2, EEF1A1, PTMA, HLA-DR, C6orf115, PBX3, ENPP4, SELL, and IL3Ra were expressed more than 2-fold higher in B-lymphoid compared to myeloid cell lines studied here. Conclusion: Thus, B-lymphoid leukaemia cell lines here exhibited active, proliferating characteristics closer to GP genes. Higher expression of several genes in B-lymphoid compared to myeloid leukaemia cell lines may be useful markers to study biological differences including drug resistance between lineages.

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