NDRG1 is down-regulated in the early apoptotic event induced by camptothecin analogs: The potential role in proteolytic activation of PKCδ and apoptosis

• Published in: Proteomics (Weinheim) 2009-04, Vol.9 (8), p.2064-2075
• Main Authors: Zheng, Ying, Wang, Li-Shun, Xia, Li, Han, Yu-Hui, Liao, Shi-Hua, Wang, Xiao-Ling, Cheng, Jin-Ke, Chen, Guo-Qiang
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Antibiotics, Antineoplastic - pharmacology; Antineoplastic Agents, Phytogenic - pharmacology; Apoptosis; Apoptosis - drug effects; Apoptosis - genetics; Biological and medical sciences; Camptothecin; Camptothecin - analogs & derivatives; Camptothecin - pharmacology; Cell Cycle Proteins - genetics; Cell Cycle Proteins - metabolism; Down-Regulation; Doxorubicin - pharmacology; Enzyme Activation - drug effects; Fundamental and applied biological sciences. Psychology; Humans; Intracellular Signaling Peptides and Proteins - genetics; Intracellular Signaling Peptides and Proteins - metabolism; Miscellaneous; N-myc downstream regulated gene 1; Protein kinase C delta; Protein Kinase C-delta - metabolism; Proteins; RNA, Small Interfering - metabolism; Topotecan - pharmacology; Transcription, Genetic - drug effects; U937 Cells
• ISSN: 1615-9853; 1615-9861
• DOI: 10.1002/pmic.200800031

We previously reported that NSC606985, a new camptothecin analog, induces apoptosis of acute myeloid leukemic cells, which is triggered by proteolytic activation of protein kinase C delta (PKCδ). Here, we performed quantitative proteomic analysis of NSC606985-treated and untreated leukemic U937 cells with two-dimensional fluorescence difference gel electrophoresis (2-D DIGE) in combination with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight tandem mass spectrometry. Thirty-three proteins were found to be deregulated. Then, we focused on N-myc downstream regulated gene 1 (NDRG1) down-regulated during apoptosis induction. The results demonstrated that the down-regulation of NDRG1 protein but not its mRNA was an early event prior to proteolytic activation of PKCδ in U937 cells under treatments of NSC606985 as well as other camptothecin analogs. With the ectopic expression of NDRG1, the proteolytic activation of PKCδ in NSC606985-treated leukemic cells was delayed and the cells were less sensitive to apoptosis. On the contrary, the suppression of NDRG1 expression by specific small interfering RNA significantly enhanced NSC606985-induced activation of PKCδ and apoptosis of U937 cells. In summary, our study suggests that the down-regulation of NDRG1 is involved in proteolytic activation of PKCδ during apoptosis induction, which would shed new light on the understanding the apoptotic process initiated by camptothecin.