Introduction <p>Leukemia, like most cancers, likely originates from a unique genetic defect. Accessible models that accurately reflect this uniqueness for studying cellular characteristics and responses are still lacking. The <i>KMT2A/MLLT1</i> fusion gene (encoding the KMT2A/ENL fusion protein) is a common chromosomal translocation in acute leukemia and demonstrates strong oncogenic potential. <i>KMT2A</i> is also referred to as mixed lineage leukemia (<i>MLL</i>).</p> Objective <p>The aim of this study was to generate leukemia-like B-cells from human adult cells by transfecting KMT2A/MLLT1 fusion gene into healthy PBMCs.</p> Method <p>The <i>KMT2A/MLLT1</i> fusion gene was inserted into peripheral blood mononuclear cells (PBMCs) isolated from three healthy individuals. Triplicate experiments were performed using the lipofection method. Flow cytometry confirmed induced cells by detecting expression of the KMT2A/ENL (MLL +) fusion protein and characterized B-cell leukemia markers (CD19, CD34, CD45 and CD38).</p> Results <p>MLL + CD45 + cells significantly increased to 45.7 ± 19.5% (N = 9) in transfected cells on day 10. The mean fluorescence intensity of CD45, CD34 and CD38, however, remained unchanged. Interestingly, unlike untransfected cells, the percentage of MLL + CD19 + (CD45 + CD38 +) B cells also significantly increased by day 10 (35.4 ± 32.0% vs 3.7 ± 2.4%) demonstrating abnormal growth characteristics.</p> Conclusions <p>These rapid, accessible and reproducible KMT2A/ENL + leukemia-like cells may serve as a suitable model for studying leukemia.</p>

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Altered growth in KMT2A/ENL induced human B cells from peripheral blood

  • Siti Zuleha Idris,
  • Vivek Prasad Jayamoney,
  • Amrina Mohd Amin,
  • Sethu Thakanachy Subha,
  • Mei I Lai,
  • Fazlina Nordin,
  • Maha Abdullah

摘要

Introduction

Leukemia, like most cancers, likely originates from a unique genetic defect. Accessible models that accurately reflect this uniqueness for studying cellular characteristics and responses are still lacking. The KMT2A/MLLT1 fusion gene (encoding the KMT2A/ENL fusion protein) is a common chromosomal translocation in acute leukemia and demonstrates strong oncogenic potential. KMT2A is also referred to as mixed lineage leukemia (MLL).

Objective

The aim of this study was to generate leukemia-like B-cells from human adult cells by transfecting KMT2A/MLLT1 fusion gene into healthy PBMCs.

Method

The KMT2A/MLLT1 fusion gene was inserted into peripheral blood mononuclear cells (PBMCs) isolated from three healthy individuals. Triplicate experiments were performed using the lipofection method. Flow cytometry confirmed induced cells by detecting expression of the KMT2A/ENL (MLL +) fusion protein and characterized B-cell leukemia markers (CD19, CD34, CD45 and CD38).

Results

MLL + CD45 + cells significantly increased to 45.7 ± 19.5% (N = 9) in transfected cells on day 10. The mean fluorescence intensity of CD45, CD34 and CD38, however, remained unchanged. Interestingly, unlike untransfected cells, the percentage of MLL + CD19 + (CD45 + CD38 +) B cells also significantly increased by day 10 (35.4 ± 32.0% vs 3.7 ± 2.4%) demonstrating abnormal growth characteristics.

Conclusions

These rapid, accessible and reproducible KMT2A/ENL + leukemia-like cells may serve as a suitable model for studying leukemia.