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Updated: Jul 13, 2026
Intracellular Phosphoflow Cytometry of Acute Myeloid Leukemia Patient-Derived Xenotransplants
Published on: June 6, 2025
Subcellular proteome analysis of camptothecin analogue NSC606985-treated acute myeloid leukemic cells
Yun Yu1, Li-Shun Wang, Shao-Ming Shen
1Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Rui-Jin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Abstract:
We reported previously that NSC606985, a camptothecin analogue, induces apoptosis of acute myeloid leukemia (AML) cells through proteolytic activation of protein kinase Cdelta. Here, we analyzed protein expression profiles of fractionated nuclei, mitochondria, raw endoplasmic reticula, and cytosols of NSC606985-induced apoptotic AML cell line NB4 cells by two-dimensional electrophoresis combined with MALDI-TOF/TOF tandem mass spectrometry. In total, 90 unique deregulated proteins, including 16 compartment-compartment translocated ones, were identified. They contributed to multiple functional activities such as DNA damage repairing, chromosome assembly, mRNA processing, biosynthesis, modification, and degradation of proteins. More interestingly, several increased oxidative stress-related proteins mainly presented in mitochondria, while upregulated glycolysis proteins mainly occurred in the nuclei. With their functional analyses, the possible roles of these deregulated proteins in NSC606985-induced apoptosis were discussed. Collectively, these discoveries would shed new insights for systematically understanding the mechanisms of the camptothecin-induced apoptosis.
Insights
NSC606985 triggers cancer cell death by altering protein functions within cellular compartments. This study identifies key proteins involved in DNA repair, stress responses, and energy metabolism during apoptosis.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- NSC606985, a camptothecin analogue, induces apoptosis in acute myeloid leukemia (AML) cells.
- Previous research indicated proteolytic activation of protein kinase Cdelta as a mechanism.
Purpose of the Study:
- To comprehensively analyze protein expression profiles in subcellular compartments of apoptotic AML cells.
- To identify deregulated proteins and their functional roles in NSC606985-induced apoptosis.
Main Methods:
- Utilized two-dimensional electrophoresis and MALDI-TOF/TOF tandem mass spectrometry.
- Analyzed protein expression in fractionated nuclei, mitochondria, endoplasmic reticula, and cytosols.
- Identified 90 unique deregulated proteins, including 16 showing compartment translocation.
Main Results:
- Identified proteins involved in DNA repair, chromosome assembly, mRNA processing, and protein metabolism.
- Observed increased oxidative stress-related proteins in mitochondria.
- Found upregulated glycolysis proteins predominantly in the nuclei.
Conclusions:
- Discovered novel insights into the mechanisms of camptothecin-induced apoptosis.
- Highlighted the differential localization and function of deregulated proteins in apoptosis.
- Provided a foundation for understanding NSC606985's effects on AML cells.
