Truncated apolipoprotein C-I induces apoptosis in neuroblastoma by activating caspases in the extrinsic and intrinsic pathways.

Truncated apolipoprotein C-I is a post-translationally modified protein characterized by the loss of threonine and proline residues from the N-terminus of the mature peptide. The truncated peptide is involved in many physiological and pathological processes in vivo and is related to malignant diseases. The aim of the present study was to assess the effects of the truncated peptide on tumorigenesis in neuroblastoma. The truncated peptide was chemically synthesized, and a signal peptide was used as the negative control. The results of the CCK-8 assay showed that the truncated peptide selectively inhibited cell proliferation compared with the signal peptide, and inhibited migration and invasion as determined by wound healing and Transwell assays. Flow cytometry analysis demonstrated that the truncated peptide induced apoptosis and cell cycle arrest in the S phase. Bax, Bim, and tBid upregulation, and Bcl‑2 and Bcl‑xl downregulation were associated with permeabilization of the mitochondrial membrane, as detected by the JC-1 assay and the release of cytochrome c and apoptosis. Activation of caspase‑8 was associated with activation of cell death receptors such as the tumor necrosis factor receptor. PARP cleavage indicated apoptosis, and DNA damage was observed in the TUNEL assay. The results showed that the truncated apoC-I induced apoptosis in neuroblastoma by the extrinsic and intrinsic pathways. The anticancer effects were confirmed in vivo in a xenograft mouse model. In conclusion, the endogenous protein apoC-I may be a new promising therapeutic target to suppress tumor growth.

[1]  R. Wolterbeek,et al.  Apolipoprotein C‐I plays a role in the pathogenesis of glomerulosclerosis , 2017, The Journal of pathology.

[2]  C. Van Waes,et al.  Therapeutic Small Molecules Target Inhibitor of Apoptosis Proteins in Cancers with Deregulation of Extrinsic and Intrinsic Cell Death Pathways , 2016, Clinical Cancer Research.

[3]  J. Dou,et al.  Wip1 inhibitor GSK2830371 inhibits neuroblastoma growth by inducing Chk2/p53-mediated apoptosis , 2016, Scientific Reports.

[4]  H. Juan,et al.  A multidisciplinary team care approach improves outcomes in high-risk pediatric neuroblastoma patients , 2016, Oncotarget.

[5]  Zhenghu Chen,et al.  Novel multi-targeted ErbB family inhibitor afatinib blocks EGF-induced signaling and induces apoptosis in neuroblastoma , 2016, Oncotarget.

[6]  H. Ko,et al.  Apolipoprotein C1 (APOC1) as a novel diagnostic and prognostic biomarker for lung cancer: A marker phase I trial , 2014, Thoracic cancer.

[7]  Shu Zheng,et al.  Identification of Apolipoprotein C-I as a Potential Wilms’ Tumor Marker after Excluding Inflammatory Factors , 2014, International journal of molecular sciences.

[8]  D. L. Puppione.  Higher primates, but not New World monkeys, have a duplicate set of enhancers flanking their apoC-I genes. , 2014, Comparative biochemistry and physiology. Part D, Genomics & proteomics.

[9]  Feng Zhang,et al.  Tetramethylpyrazine induces G0/G1 cell cycle arrest and stimulates mitochondrial-mediated and caspase-dependent apoptosis through modulating ERK/p53 signaling in hepatic stellate cells in vitro , 2013, Apoptosis.

[10]  T. Han,et al.  HCV NS4B induces apoptosis through the mitochondrial death pathway. , 2012, Virus research.

[11]  Wan-ying Wu,et al.  Cardio-Protection of Salvianolic Acid B through Inhibition of Apoptosis Network , 2011, PloS one.

[12]  Shengshou Hu,et al.  Apoptosis of mesenchymal stem cells induced by hydrogen peroxide concerns both endoplasmic reticulum stress and mitochondrial death pathway through regulation of caspases, p38 and JNK , 2010, Journal of cellular biochemistry.

[13]  X. Xiao,et al.  Detection of two distinct forms of apoC-I in great apes. , 2010, Comparative biochemistry and physiology. Part D, Genomics & proteomics.

[14]  A. Strasser,et al.  The many roles of FAS receptor signaling in the immune system. , 2009, Immunity.

[15]  T. Tomonaga,et al.  The isolation and identification of apolipoprotein C-I in hormone-refractory prostate cancer using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry. , 2009, Asian journal of andrology.

[16]  M. Miyazaki,et al.  Apolipoprotein C-1 maintains cell survival by preventing from apoptosis in pancreatic cancer cells , 2008, Oncogene.

[17]  J. Romijn,et al.  Apolipoprotein C-I binds free fatty acids and reduces their intracellular esterification Published, JLR Papers in Press, March 5, 2007. , 2007, Journal of Lipid Research.

[18]  R. Kleemann,et al.  Apolipoprotein CI stimulates the response to lipopolysaccharide and reduces mortality in Gram‐negative sepsis , 2006, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[19]  S. Cook,et al.  Activation of the ERK1/2 Signaling Pathway Promotes Phosphorylation and Proteasome-dependent Degradation of the BH3-only Protein, Bim* , 2003, Journal of Biological Chemistry.

[20]  K. Lei,et al.  JNK phosphorylation of Bim-related members of the Bcl2 family induces Bax-dependent apoptosis , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[21]  S. Cory,et al.  The Bcl2 family: regulators of the cellular life-or-death switch , 2002, Nature Reviews Cancer.

[22]  D. Tang,et al.  Identification and characterization of Bimgamma, a novel proapoptotic BH3-only splice variant of Bim. , 2002, Cancer research.

[23]  A. Strasser,et al.  BH3-only Bcl-2 family member Bim is required for apoptosis of autoreactive thymocytes , 2002, Nature.

[24]  S. Korsmeyer,et al.  Proapoptotic BAX and BAK: A Requisite Gateway to Mitochondrial Dysfunction and Death , 2001, Science.

[25]  O. Bernard,et al.  Dominant-Negative c-Jun Promotes Neuronal Survival by Reducing BIM Expression and Inhibiting Mitochondrial Cytochrome c Release , 2001, Neuron.

[26]  S. Korsmeyer,et al.  Pro-apoptotic cascade activates BID, which oligomerizes BAK or BAX into pores that result in the release of cytochrome c , 2000, Cell Death and Differentiation.

[27]  X. Liu,et al.  An APAF-1·Cytochrome c Multimeric Complex Is a Functional Apoptosome That Activates Procaspase-9* , 1999, The Journal of Biological Chemistry.

[28]  P. Bondarenko,et al.  Mass spectral study of polymorphism of the apolipoproteins of very low density lipoprotein. , 1999, Journal of lipid research.

[29]  A. Strasser,et al.  Bim: a novel member of the Bcl‐2 family that promotes apoptosis , 1998, The EMBO journal.

[30]  T. Nagatsu,et al.  Activity of dipeptidyl peptidase IV and post-proline cleaving enzyme in sera from osteoporotic patients. , 1988, Clinical chemistry.

[31]  J. Scott,et al.  Characterisation of mRNAs encoding the precursor for human apolipoprotein CI. , 1984, Nucleic acids research.

[32]  T. Morimoto,et al.  Mechanisms for the incorporation of proteins in membranes and organelles , 1982, The Journal of cell biology.

[33]  D. Fredrickson,et al.  Thf complete amino acid sequence of C-I (apoLp-Ser), an apolipoprotein from human very low density lipoproteins. , 1975, The Journal of biological chemistry.

[34]  A. Gotto,et al.  The primary structure of apolopoprotein-serine. , 1974, The Journal of biological chemistry.

[35]  Yoshio Kobayashi,et al.  The effects of DPP-4 inhibitor on hypoxia-induced apoptosis in human umbilical vein endothelial cells. , 2017, Journal of pharmacological sciences.

[36]  L. Falstrault,et al.  Apolipoprotein C-I reduces cholesteryl esters selective uptake from LDL and HDL by binding to HepG2 cells and lipoproteins. , 2010, Biochimica et biophysica acta.

[37]  K. Murphy,et al.  Bcl-2 inhibits Bax translocation from cytosol to mitochondria during drug-induced apoptosis of human tumor cells , 2000, Cell Death and Differentiation.