Inhibition of neuroblastoma proliferation by PF-3758309, a small-molecule inhibitor that targets p21-activated kinase 4

Neuroblastoma is the most common extracranial solid childhood tumor. Despite the availability of advanced multimodal therapy, high-risk patients still have low survival rates. p21-activated kinase 4 (PAK4) has been shown to regulate many cellular processes in cancer cells, including migration, polarization and proliferation. However, the role of PAK4 in neuroblastoma remains unclear. In the present study, we demonstrated that PAK4 was overexpressed in neuroblastoma tissues and was correlated with tumor malignance and prognosis. To investigate the function of PAK4 in neuroblastoma, we used a small-molecule inhibitor that targets PAK4, that is, PF-3758309. Our results showed that PF-3758309 significantly induced cell cycle arrest at the G1 phase and apoptosis in neuroblastoma cell lines. Meanwhile, the inhibition of PAK4 by PF-3758309 increased the expression of CDKN1A, BAD and BAK1 and decreased the expression of Bcl-2 and Bax. In addition, we screened the target genes of PAK4 by PCR array and found that 23 genes were upregulated (including TP53I3, TBX3, EEF1A2, CDKN1A, IFNB1 and MAPK8IP2) and 20 genes were downregulated (including TNFSF8, Bcl2-A1, Bcl2L1, SOCS3, BIRC3 and NFKB1) after PAK4 inhibition by PF-3758309. Moreover, PAK4 was found to regulate the cell cycle and apoptosis via the ERK signaling pathway. In conclusion, the present study demonstrated, for the first time, the expression and function of PAK4 in neuroblastomas and the inhibitory effect of PF-3758309, which deserves further investigation as an alternative strategy for neuroblastoma treatment.

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[38]  O. Bernard,et al.  Cytoskeletal Changes Regulated by the PAK4 Serine/Threonine Kinase Are Mediated by LIM Kinase 1 and Cofilin* , 2001, The Journal of Biological Chemistry.

[39]  N. Gnesutta,et al.  The Serine/Threonine Kinase PAK4 Prevents Caspase Activation and Protects Cells from Apoptosis* , 2001, The Journal of Biological Chemistry.

[40]  A. Abo,et al.  PAK4, a novel effector for Cdc42Hs, is implicated in the reorganization of the actin cytoskeleton and in the formation of filopodia , 1998, The EMBO journal.

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[43]  D. Cheresh,et al.  Glioblastomas require integrin αvβ3/PAK4 signaling to escape senescence. , 2015, Cancer research.

[44]  He Sun,et al.  PAK4 confers the malignance of cervical cancers and contributes to the cisplatin-resistance in cervical cancer cells via PI3K/AKT pathway , 2015, Diagnostic Pathology.

[45]  Ye Zhang,et al.  Activated Pak4 expression correlates with poor prognosis in human gastric cancer patients , 2015, Tumor Biology.

[46]  I. Ellis,et al.  Small molecule inhibition of group I p21-activated kinases in breast cancer induces apoptosis and potentiates the activity of microtubule stabilizing agents , 2015, Breast Cancer Research.

[47]  Peng-Sheng Zheng,et al.  CIP2A cooperates with H-Ras to promote epithelial-mesenchymal transition in cervical-cancer progression. , 2015, Cancer letters.

[48]  Liya Liu,et al.  MicroRNA-433 inhibits cell proliferation in hepatocellular carcinoma by targeting p21 activated kinase (PAK4) , 2014, Molecular and Cellular Biochemistry.

[49]  A. Singh,et al.  p-21 activated kinase 4 promotes proliferation and survival of pancreatic cancer cells through AKT- and ERK-dependent activation of NF-κB pathway , 2014, Oncotarget.

[50]  Liu Cao,et al.  Oncogenic PAK4 regulates Smad2/3 axis involving gastric tumorigenesis , 2014, Oncogene.

[51]  Jiarui Feng,et al.  PAK4 confers cisplatin resistance in gastric cancer cells via PI3K/Akt- and MEK/Erk-dependent pathways. , 2014, Bioscience reports.

[52]  Jongsung Lee,et al.  PF-3758309, p21-activated kinase 4 inhibitor, suppresses migration and invasion of A549 human lung cancer cells via regulation of CREB, NF-κB, and β-catenin signalings , 2013, Molecular and Cellular Biochemistry.

[53]  J. Chernoff,et al.  PAK signalling during the development and progression of cancer , 2013, Nature Reviews Cancer.

[54]  Armand Bankhead,et al.  Kinase pathway dependence in primary human leukemias determined by rapid inhibitor screening. , 2013, Cancer research.

[55]  A. Minden.  The Pak4 Protein Kinase in Breast Cancer , 2012, ISRN oncology.

[56]  M. Gujrati,et al.  Functional cooperativity by direct interaction between PAK4 and MMP-2 in the regulation of anoikis resistance, migration and invasion in glioma , 2012, Cell Death and Disease.

[57]  W. T. Schaiff,et al.  Evaluation of Potential Gastrointestinal Biomarkers in a PAK4 Inhibitor-treated Preclinical Toxicity Model to Address Unmonitorable Gastrointestinal Toxicity , 2012, Toxicologic pathology.

[58]  A. Minden,et al.  PAK4 is required for regulation of the cell‐cycle regulatory protein p21, and for control of cell‐cycle progression , 2011, Journal of cellular biochemistry.

[59]  A. Cheung,et al.  Overexpressed PAK4 promotes proliferation, migration and invasion of choriocarcinoma. , 2011, Carcinogenesis.

[60]  S. Tsao,et al.  p21-activated kinase 4 regulates ovarian cancer cell proliferation, migration, and invasion and contributes to poor prognosis in patients , 2010, Proceedings of the National Academy of Sciences.

[61]  S. Strömblad,et al.  Integrin-mediated Cell Attachment Induces a PAK4-dependent Feedback Loop Regulating Cell Adhesion through Modified Integrin αvβ5 Clustering and Turnover , 2010, Molecular biology of the cell.

[62]  E. Manser,et al.  Do PAKs make good drug targets? , 2010, F1000 biology reports.

[63]  N. Chen,et al.  The protein kinase Pak4 disrupts mammary acinar architecture and promotes mammary tumorigenesis , 2010, Oncogene.

[64]  J. Maris.  Recent advances in neuroblastoma. , 2010, The New England journal of medicine.

[65]  J. Masters,et al.  PAK4: a pluripotent kinase that regulates prostate cancer cell adhesion , 2010, Journal of Cell Science.

[66]  D. Knighton,et al.  Small-molecule p21-activated kinase inhibitor PF-3758309 is a potent inhibitor of oncogenic signaling and tumor growth , 2010, Proceedings of the National Academy of Sciences.

[67]  Hang Zheng,et al.  [Expression of PAK4 in breast cancer and benign breast pathological changes]. , 2010, Nan fang yi ke da xue xue bao = Journal of Southern Medical University.

[68]  A. Minden,et al.  Essential role for the Pak4 protein kinase in extraembryonic tissue development and vessel formation , 2009, Mechanisms of Development.

[69]  M. Naujokas,et al.  Pak4, a Novel Gab1 Binding Partner, Modulates Cell Migration and Invasion by the Met Receptor , 2009, Molecular and Cellular Biology.

[70]  J. Kendall,et al.  Copy number alterations in pancreatic cancer identify recurrent PAK4 amplification , 2008, Cancer biology & therapy.

[71]  A. Minden,et al.  The Pak4 Protein Kinase Plays a Key Role in Cell Survival and Tumorigenesis in Athymic Mice , 2008, Molecular Cancer Research.

[72]  A. Minden,et al.  Pak4 Induces Premature Senescence via a Pathway Requiring p16INK4/p19ARF and Mitogen-Activated Protein Kinase Signaling , 2005, Molecular and Cellular Biology.

[73]  A. Minden,et al.  PAK4 Functions in Tumor Necrosis Factor (TNF) α-induced Survival Pathways by Facilitating TRADD Binding to the TNF Receptor* , 2005, Journal of Biological Chemistry.

[74]  G. Bokoch.  Biology of the p21-activated kinases. , 2003, Annual review of biochemistry.

[75]  R. Bronson,et al.  PAK4 Kinase Is Essential for Embryonic Viability and for Proper Neuronal Development , 2003, Molecular and Cellular Biology.

[76]  G. Brodeur.  Neuroblastoma: biological insights into a clinical enigma , 2003, Nature Reviews Cancer.

[77]  D. Whyte,et al.  Requirement for PAK4 in the Anchorage-independent Growth of Human Cancer Cell Lines* , 2002, The Journal of Biological Chemistry.

[78]  O. Bernard,et al.  Cytoskeletal Changes Regulated by the PAK4 Serine/Threonine Kinase Are Mediated by LIM Kinase 1 and Cofilin* , 2001, The Journal of Biological Chemistry.

[79]  N. Gnesutta,et al.  The Serine/Threonine Kinase PAK4 Prevents Caspase Activation and Protects Cells from Apoptosis* , 2001, The Journal of Biological Chemistry.

[80]  A. Abo,et al.  PAK4, a novel effector for Cdc42Hs, is implicated in the reorganization of the actin cytoskeleton and in the formation of filopodia , 1998, The EMBO journal.