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Modeling Chemotherapy Resistant Leukemia In Vitro
Published on: February 9, 2016
PRAME promotes in vitro leukemia cells death by regulating S100A4/p53 signaling
1Department of Haematology, People's Hospital of Linyi, Shandong, China. xypgj@sina.com.
Objective:
PRAME (Preferentially Expressed Antigen in Melanoma) is a tumor-associated antigen recognized by immunocytes, and it induces cytotoxic T cell-mediated responses in melanoma. PRAME is expressed in a wide variety of tumors, but in contrast with most other tumor-associated antigens, it is also expressed in leukemias. The physiologic role of PRAME remains elusive. Recently, it has found PRAME could be involved in the regulation of cell death in leukemias, but the mechanism of the function is unclear. Here, we confirm that PRAME induces leukemias cell death by regulation of S100A4/p53 signaling.
Materials And Methods:
The pCDNA3-PRAME plasmid and its control were transfected with the KG-1 cells. The pCDNA3-PRAME transfected KG-1 cells were then transiently transfected with S100A4 cDNA or wt-p53 siRNA. The PRAME siRNA and its control were transfected with the K562 cells. The PRAME siRNA transfected K562 cells were then transiently transfected with S100A4 siRNA or pGMp53-Lu. PRAME, S100A4 and P53 were detected by Western blot assay in different time point. Annexin V/propidium iodide and MTT methods were used to detect apoptosis and cell survival rate.
Results:
KG-1 cells overexpressing the PRAME gene significantly induces apoptosis and decreases proliferation in vitro, followed by down-regulation of S100A4 and up-regulation of p53. Up-regulation of S100A4 by S100A4 transfection inhibits PRAME-induced p53 up-regulation. Furthermore, up-regulation of S100A4 by S100A4 transfection or down-regulation of p53 by p53 siRNA transfection reduces apoptosis and increases proliferation in vitro. Knockdown of PRAME in K562 cells significantly increases proliferation in vitro, followed by up-regulation of S100A4 and down-regulation of p53. The downregulation of S100A4 by S100A4 siRNA transfection increased p53 expression. Furthermore, downregulation of S100A4 by S100A4 siRNA transfection or up-regulation of p53 by p53 transfection decreases proliferation in vitro.
Conclusions:
Our results suggest that the leukemias expressing high levels of PRAME has a favorable prognosis. PRAME promotes in vitro leukemia cells death by regulating S100A4/p53 signaling.
Insights
Preferentially Expressed Antigen in Melanoma (PRAME) promotes leukemia cell death by regulating the S100A4/p53 signaling pathway. High PRAME expression in leukemia indicates a favorable prognosis, suggesting its therapeutic potential.
Area of Science:
- Oncology
- Molecular Biology
- Immunology
Background:
- Preferentially Expressed Antigen in Melanoma (PRAME) is a tumor antigen involved in cytotoxic T cell responses.
- PRAME expression is observed in various tumors, including leukemias, but its precise role in leukemia pathogenesis is unclear.
- Previous studies suggest PRAME's involvement in regulating cell death in leukemias, yet the underlying mechanisms require elucidation.
Purpose of the Study:
- To investigate the mechanism by which PRAME influences leukemia cell death.
- To determine the role of the S100A4/p53 signaling pathway in PRAME-mediated apoptosis.
- To assess the prognostic significance of PRAME expression in leukemia.
Main Methods:
- Gene transfection and siRNA techniques were employed to manipulate PRAME, S100A4, and p53 expression in KG-1 and K562 leukemia cell lines.
- Western blot assays were utilized to detect protein expression levels of PRAME, S100A4, and p53.
- Apoptosis was quantified using Annexin V/propidium iodide staining, and cell proliferation was assessed via MTT assays.
Main Results:
- Overexpression of PRAME in KG-1 cells induced apoptosis and reduced proliferation, accompanied by decreased S100A4 and increased p53 levels.
- Modulation of S100A4 or p53 expression affected PRAME-induced apoptosis and proliferation, highlighting their regulatory roles.
- Knockdown of PRAME in K562 cells led to increased proliferation, elevated S100A4, and reduced p53, with subsequent interventions reversing these effects.
Conclusions:
- PRAME facilitates in vitro leukemia cell death through the regulation of S100A4/p53 signaling.
- Leukemia patients with high PRAME expression exhibit a favorable prognosis.
- These findings elucidate PRAME's function in leukemia and suggest its potential as a prognostic marker and therapeutic target.
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