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[p73 gene expression in apoptotic process of acute myeloid leukemia cell line U937 induced by methotrexate]

Guang-Fen Xiao1, Qing Lu, Xiang-Dong Yang

  • 1Department of Hematology, The Second Affiliated Hospital of Nanhua University, Hengyang 421001, China.

Zhongguo Shi Yan Xue Ye Xue Za Zhi
|January 7, 2003
PubMed

Insights

Methotrexate effectively induces apoptosis in acute myeloid leukemia U937 cells. However, this process does not alter the expression of the p73 gene mRNA in these cancer cells.

Area of Science:

  • Molecular Biology
  • Cancer Research
  • Cell Biology

Background:

  • Acute myeloid leukemia (AML) is a heterogeneous hematological malignancy.
  • Understanding the molecular mechanisms of apoptosis in AML is crucial for developing targeted therapies.
  • The p73 gene is a tumor suppressor and plays a role in apoptosis.

Purpose of the Study:

  • To investigate the role of p73 gene expression during methotrexate-induced apoptosis in the U937 AML cell line.
  • To analyze the impact of methotrexate on cell morphology, DNA integrity, and cell cycle progression in U937 cells.

Main Methods:

  • U937 cells were treated with methotrexate (MTX).
  • Apoptosis was assessed via microscopy, Wright's + Giemsa staining, and DNA laddering (agarose gel electrophoresis).
  • Cell cycle analysis was performed using flow cytometry (FCM), and p73 mRNA expression was quantified using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR).

Main Results:

  • Methotrexate effectively induced apoptosis in U937 cells, evidenced by morphological changes like condensed nuclei and chromosome fragmentation.
  • DNA laddering and a Sub-G1 peak in FCM indicated DNA damage and apoptosis.
  • Cell cycle arrest was observed in the S and G2/M phases.
  • Crucially, p73 mRNA expression levels remained constant throughout the apoptotic process.

Conclusions:

  • Methotrexate is an effective inducer of apoptosis in the U937 AML cell line.
  • The apoptotic process triggered by methotrexate in U937 cells does not involve alterations in p73 mRNA levels.
  • These findings suggest that p73 gene expression is not a key regulator in MTX-induced apoptosis in this AML model.

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