An Active Compound from the Pyrazine Family Induces Apoptosis by Targeting the Bax/Bcl2 and Survivin Expression in Chronic Myeloid Leukemia K562 Cells

Author(s):

Saeedeh Rostampour, Farhad Eslami, Esmaeil Babaei, Hossein Mostafavi and Majid Mahdavi*   Pages 203 - 212 ( 10 )

Abstract:

<p>Background: It has been established that pyrazine derivatives, which have widespread bioactivities, can effectively treat cancer. <p> Objectives: In this study, we investigated the effects of <i>2-methoxy-5-(oxiran-2-ylmethyl) phenyl pyrazine-2- carboxylate</i> (2-mOPP), a new pyrazine derivative, on proliferation, viability, and apoptosis induction in human leukemia K562 cells. <p> Methods: For this purpose, the K562 cells were treated with various concentrations (20-120 μM) of the 2-mOPP for 24-72 hours. Cell viability was determined by MTT growth inhibition assay. Apoptotic activity of 2-mOPP was investigated morphologically by Hoechst staining, cell surface expression assay of phosphatidylserine by Annexin-V/PI technique, as well as DNA fragmentation assay. The effect of 2-mOPP on the K562 cell cycle was studied by flow cytometry. To determine the impact of 2-mOPP on the expression of intrinsic apoptosis-related genes, Bcl2 (anti-apoptotic), Bax (pro-apoptotic), and Survivin genes expression levels were evaluated before and after treatment with 2-mOPP through Real-Time PCR analysis. <p> Results: The results revealed that 2-mOPP inhibited viability with IC₅₀ of 25μM in 72 h. Morphological changes assessment by fluorescence microscopy, Annexin V/PI double staining by flow cytometry, and DNA ladders formation upon cell treatment with the 2-mOPP showed that this compound induces apoptosis at IC₅₀ value. Cell cycle arrest was observed in the G₀/G₁ phase, and the sub-G₁ cell population (the sign of apoptosis) increased in a time-dependent manner. Low expression levels of Bcl2 and Survivin in K562 cells were observed 24-72 h after treatment. Along with the down-regulation of Survivin and Bcl2, the expression of Bax was increased after treatment with 2-mOPP. <p> Conclusion: These findings demonstrate that the new pyrazine derivative plays a crucial role in blocking the proliferation of the leukemic cells by inducing cell cycle arrest and apoptosis.</p>

Keywords:

2-mOPP, apoptosis, leukemia, Bcl2, survivin, K562 cell.

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