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Preparation of Primary Acute Lymphoblastic Leukemia Cells in Different Cell Cycle Phases by Centrifugal Elutriation
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[The Study of PI3K-Ⅲ Like Functional Polypeptide on Leukemia Cell K562 during the Process of Programmed Cell Death]

Ben Liu1, Wen Dong1, Jie Sun1

  • 1Pharmaceutical and Medical Technology College, Key Laboratory of Medical Microecology (Putian University), Fujian Province University.

Zhongguo Shi Yan Xue Ye Xue Za Zhi
|August 18, 2022
PubMed
Abstract

Insights

The PI3K-III like functional domain protein induces programmed cell death in K562 leukemia cells by inhibiting proliferation and promoting apoptosis. This process involves the caspase pathway and may coordinate autophagy and apoptosis.

Area of Science:

  • Molecular Biology
  • Cell Biology
  • Biochemistry

Background:

  • Leukemia remains a significant health challenge, necessitating research into novel therapeutic targets.
  • Understanding programmed cell death mechanisms is crucial for developing effective anti-cancer strategies.
  • The PI3K-III pathway plays a role in cellular processes, but its specific function in leukemia cell death requires further elucidation.

Purpose of the Study:

  • To investigate the molecular mechanisms by which the PI3K-III like functional domain induces programmed cell death in the K562 leukemia cell line.
  • To determine the role of autophagy and apoptosis in this process.

Main Methods:

  • Protein expression and purification of the PI3K-III like functional domain using the Pichia pastoris system.
  • Assessing K562 cell proliferation and colony formation using MTT and colony-forming assays.
  • Analyzing apoptosis, cell cycle, and ultrastructural changes via flow cytometry and transmission electron microscopy.
  • Quantifying caspase-3 expression using ELISA and evaluating autophagy-related proteins (ATG4B, Beclin-1, Bcl-2, LC3-II) with Western blot.

Main Results:

  • The PI3K-III like functional domain protein significantly inhibited K562 cell proliferation and colony formation.
  • Flow cytometry revealed increased apoptosis and autophagosome formation, with cell cycle arrest in the G0/G1 phase.
  • Caspase-3 expression was significantly upregulated in a concentration-dependent manner.
  • Western blot analysis indicated dynamic changes in autophagy markers (ATG4B, Beclin-1, LC3-II) and the apoptosis regulator Bcl-2.

Conclusions:

  • The PI3K-III like functional polypeptide effectively induces programmed cell death in K562 leukemia cells.
  • The Beclin-1/Bcl-2 and caspase pathways are implicated in this cell death induction.
  • Autophagy and apoptosis appear to function concurrently in mediating the anti-leukemic effects of the PI3K-III like functional domain.
Keywords:
PI3K-Ⅲapoptosisautophagyleukemia

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