[Frontiers in Bioscience E3, 421-433, January 1, 2011]
MYCN, neuroblastoma and focal adhesion kinase (FAK)
Elizabeth A. Beierle1
1
TABLE OF CONTENTS
1. ABSTRACT
Neuroblastoma is the most common extracranial solid tumor of childhood. This tumor is characterized by poor survival, especially when it features amplification of the MYCN oncogene. The ability for human cancers to propagate is marked by their ability to invade and metastasize to distant sites. Focal adhesion kinase (FAK) is a key tyrosine kinase involved in the survival and metastasis of a number of human tumor types. We have shown that FAK is present in human neuroblastoma and that its expression in neuroblastoma is related to the MYCN oncogene. We have also demonstrated that inhibition of FAK in neuroblastoma leads to decreased tumor cell survival. The current review addresses the relationship between the MYCN oncogene, focal adhesion kinase and neuroblastoma.
2. INTRODUCTION
In the following review, we discuss the role of focal adhesion kinase in human neuroblastoma and its relationship to the MYCN oncogene in these tumors.
3. MYCN ONCOGENE
The MYCN oncogene plays a major role in human tumorigenesis. Previous data indicate that the primary function of MYCN is as a transcription factor, known to bind to the specific DNA E-box sequence, CACGTG (1, 2). Recent evidence supports a dual role for MYCN. Murphy et al recently showed that MYCN more commonly binds to the CATGTG E-box sequence, and MYCN binding correlates with DNA hypermethylation, thereby, also functioning as a mediator of chromatin structure (3). Normal expression of MYCN is limited to embryonic brain and kidney tissues, and lymphocytes early in their differentiation (4, 5, 6). The importance of MYCN in embryogenesis is demonstrated by the fact that in mice, homozygous MYCN null is an embryonic lethal mutation, with these embryos showing a significant decrease in mature neurons of neural crest origin (7). Abnormal expression of MYCN is associated with neuroblastoma, the most common extracranial solid tumor of childhood. The exact function and gene targets of MYCN have not been completely defined (8), and a thorough investigation of which target genes are required for the proliferation and tumorigenesis of neuroblastoma have not been published (9). However, there is significant interest in this arena and some progress is being made with studies showing in vitro (10, 11, 12), and in vivo (9, 13) binding of MYCN to the genes that it regulates. For example, MYCN has been shown to bind to the gene promoters for various oncogenes involved in cellular proliferation, differentiation, and survival and to increase the expression of these targets. These genes include the proto-oncogenes high mobility group A1 (HMGA1) (14) and Pax-3 (11), the multidrug resistance-associated protein gene (MRP1) (12), ATP-binding cassette (ABC) transporters that mediate efflux of chemotherapeutic agents from cancer cells (15), and the proteins nestin (10) and livin (16). There are also a number of negative transcriptional targets for MYCN in neuroblastoma including the growth-inhibitory gene Ndrg1 (17) and leukemia inhibitory factor (LIF) (18). The MYCN protein has a number of functional regions including a basic helix-loop-helix leucine zipper motif and two highly conserved areas, Myc box I and II. These regions are important for the function of the MYCN protein however, the specific regions required for transcriptional regulation are segregated and appear to vary with different target genes (19).
4. MYCN ONCOGENE AND NEUROBLASTOMA
Neuroblastoma is the most common extracranial solid tumor of childhood. Despite many advances in both medical and surgical care, the overall survival for children presenting with advanced stage disease remains less than 30%. The strongest adverse prognostic indicator in human neuroblastoma is gene amplification of greater than ten copies of the MYCN oncogene (20). Amplification of this gene occurs in about 20% of neuroblastomas (8, 20) and is associated with both increased recurrence of disease and decreased patient survival. Numerous studies have demonstrated the importance of MYCN in neuroblastoma tumorigenicity. The level of MYCN expression has been shown to correlate with the growth and proliferation of neuroblastoma cells in vitro (4, 21, 22, 23), and the interruption of MYCN induces cellular differentiation in neuroblastoma cells. Nara and colleagues showed that after blocking MYCN, neuroblastoma cells developed multidirectional neurite extension and increased cellular and nuclear size, findings consistent with differentiation (24). Downregulation of MYCN with antisense oligonucleotides resulted in a decrease in both cellular proliferation and anchorage independent growth in the cells (4, 23). Other authors have utilized small interfering RNAs (siRNAs) to silence MYCN. Woo et al demonstrated that MYCN interference resulted in a decrease in the number of neuroblastoma cells in the S-phase of the cell cycle (25). In addition, other investigators utilizing this method to silence MYCN have shown decreased neuroblastoma cell growth and increased neuroblastoma cell apoptosis (26). Finally, transgenic mice with MYCN over-expression develop spontaneous neuroblastomas (27) that have the same histological features seen in human tumors (28).
5. FOCAL ADHESION KINASE (FAK)
Focal adhesion kinase (FAK) is a nonreceptor, cytoplasmic 125kDa protein tyrosine kinase. Initial studies revealed that both the transcription of FAK mRNA (29) and the expression of FAK protein is significantly increased in primary and metastatic breast, colon, and thyroid tumors and hepatocellular carcinoma when compared normal tissues (30-37), and that these changes occur early in tumorigenesis. Real-time PCR analysis of colorectal carcinoma and liver metastasis with matched normal colonic tissues demonstrated increased FAK mRNA abundance in the tumors and metastatic tissues compared to control tissues (38), suggesting that the increased FAK expression in human tumors occurs at the level of transcription. FAK controls a number of cell signaling pathways including motility, proliferation, viability and survival (39-42). Silencing FAK expression with small interfering RNAs resulted in decreased migration of lung cancer (43) and glioblastoma cells (44), and decreased tumor growth in ovarian (45) and prostate cancer models (46). The inhibition of FAK with antisense oligonucleotides had been shown to cause decreased growth in tumor cells (47) and FAK inhibition with a dominant-negative FAK protein (FAK-CD), inhibited cell growth in human melanoma cells (29), and in human breast cancer cell lines (30, 48).
6. MYCN, NEUROBLASTOMA AND CELL ADHESION
An association between MYCN amplification and tumorigenicity through cell adhesion, motility, and invasiveness has been implied in some studies. MYCN amplified human neuroblastoma cells have both increased cellular motility and invasiveness (49) and decreased attachment (50) compared to MYCN non-amplified cell lines, but no mechanistic explanation for these observations has been elucidated. To this end, Ma and others recently demonstrated that MYCN amplification correlates with levels of microRNA-9 (miR-9) which is associated with increased cell motility and invasiveness by regulating E-cadherin in breast cancer cells (51). More specifically to neuroblastoma, Akeson and Bernards showed that rat neuroblastoma cells transfected with a MYCN expression vector have significant reductions in mRNA and protein expression of neural cell adhesion molecule, a specific cell-cell adhesion molecule (52). In addition, other researchers have shown that the expression of integrin subunits α3 and β1 are inversely related to overexpression of the N-Myc protein in neuroblastoma (53, 54). MYCN overexpression in neuroblastoma cells in vitro also results in decreased expression of α1 integrin, leading to decreased attachment and increased migratory activity (55). In addition, Wu and others have demonstrated that FAK activity is required to promote integrin stimulated neuroblastoma motility through α5β1 but not for α4β1 integrin (56). These data provide evidence that MYCN is involved in neuroblastoma cell adhesion.
7. MYCN, NEUROBLASTOMA AND FOCAL ADHESION KINASE (FAK)
The association between MYCN amplification and cellular motility and invasiveness in neuroblastoma implies a potential relationship between focal adhesion kinase (FAK) and MYCN, since FAK is a key protein involved in cellular motility.
7.1. FAK in human neuroblastoma
Early studies relating neuroblastoma and FAK primarily involved the use of SH-SY5Y neuroblastoma cells (non-amplified MYCN) to investigate the relationship between insulin-like growth factor -1 (IGF-1) and FAK activation in their role in neuronal morphology during cellular differentiation (57, 58). These investigators showed that IGF-1 stimulated cell motility was mediated through the phosphorylation of FAK. In these studies, they utilized SH-SY5Y cells, not because of their cancer properties, but because these cells can be induced to differentiate, and are thereby a good experimental model for neuronal differentiation studies. In more recent studies, Kim and Feldman began investigating the effects of FAK dephosphorylation in neuroblastoma cell survival (59, 60). In the first study they showed that treatment of SH-EP neuroblastoma cells with mannitol resulted in the loss of FAK phosphorylation and increased cellular detachment and apoptosis which were reversible by IGF-1 (59). In a subsequent study, they reported that okadaic acid treatment of SH-EP cells resulted in the loss of FAK phosphorylation and resultant apoptosis that was not reversible with the treatment of IGF-1 (60). However, until recently, there were no data demonstrating the expression of FAK in human neuroblastoma specimens, and none to clearly show that FAK and MYCN were related in neuroblastoma. An examination of 70 human neuroblastoma specimens with various INSS stage and MYCN amplification status revealed an increase in the expression of FAK protein, as detected by immunohistochemistry, in MYCN amplified human neuroblastoma specimens compared to those tumors that were not MYCN amplified (Figure 1, panels A, B) (61). Although FAK expression did not prove to be an independent prognostic indicator in that study, FAK expression was strongly associated with those advanced stage tumors that had amplification of the MYCN oncogene (61).
7.2. MYCN regulates FAK expression in neuroblastoma
Investigations into the mechanisms of FAK regulation in neuroblastoma have been ongoing. The FAK promoter has been cloned (62) and evaluations of this promoter have demonstrated a number of binding sites present for various oncogenes, such as p53. In addition, examination of the FAK promoter has revealed E-box sequences on this promoter that are potential binding sites for MYCN (Figure 2, panel A). Further studies including electrophoretic mobility shift, chromatin immunoprecipitation (ChIP), and dual luciferase assays have shown that MYCN does bind to the FAK promoter both in vitro and in vivo, resulting in an upregulation of FAK expression (63) (Figure 2, panel B). Developing a better understanding of the effects of MYCN upon the FAK promoter in neuroblastoma impacts our evaluation of the role of FAK in tumorigenesis in other tumor types. For example, MYCN is reported to be amplified in human melanoma and sarcomas (64) and is associated with poor outcomes in these tumor types (65). FAK has also been shown to be overexpressed in human sarcoma and melanoma tumors (66, 67), and downregulation of FAK results in decreased survival in human melanoma cells (68). These data provide evidence that MYCN is potentially a transcriptional regulator of FAK in these tumor types as well, but as of yet the role of MYCN here has not been fully investigated.
7.3. FAK as a target in neuroblastoma
The biological relevance of increased FAK expression by MYCN amplified neuroblastoma cell lines has begun to be explored. In an initial study, an isogenic MYCN neuroblastoma cell line with a repressible MYCN vector (SHEP Tet-21/N) (69) was utilized. Treatment of these cell lines with siRNA to FAK resulted in a significant increase in cellular apoptosis in the MYCN overexpressing cells compared to their MYCN non-expressing counterparts (63). Additional studies using AdFAK-CD, an adenoviral construct that functions as a dominant-negative of FAK, for FAK inhibition in the same isogenic MYCN neuroblastoma cell lines resulted in decreased cellular attachment and proliferation and increased cellular apoptosis in the neuroblastoma cells that had MYCN overexpression compared to the non-overexpressing cell line (70).
Recent advances have been made in the development of small molecule inhibitors of FAK. For instance, TAE226, a small molecule FAK inhibitor, has been reported to decrease cell survival in human glioma cells (71) and to decrease metastasis and enhance survival in xenograft models of pancreatic and breast cancer (72, 73). TAE226 has been studied in neuroblastoma and found to decrease tumor cell survival (74). FAK inhibition with TAE226 in a MYCN amplified neuroblastoma cell line (SK-N-BE(2)) results in significantly decreased cell viability at low concentrations with minimal effects upon a MYCN non-amplified cell line (SK-N-AS) (Figure 3). Other small molecule FAK inhibitors are also being examined. Golubovskaya et al have reported their results with 1,2,4,5-benzenetetraamine tetrahydrochloride, a small molecule directed towards the site of autophosphorylation on FAK (75). FAK inhibition using 1,2,4,5-benzenetetraamine tetrahydrochloride resulted in decreased breast cancer cell survival both in vitro and in vivo. Other investigators have demonstrated the efficacy of this small molecule in treating pancreatic cancer cell xenografts (76). Use of 1,2,4,5-benzenetetraamine tetrahydrochloride in human neuroblastoma has also been reported (77). Treatment with 1,2,4,5-benzenetetraamine tetrahydrochloride resulted in decreased cellular attachment, viability and increased apoptosis in vitro, and decreased neuroblastoma tumor growth in vivo, in a number of human neuroblastoma cell lines. As seen in previous studies, neuroblastoma cell lines with MYCN amplification were more sensitive to FAK inhibition with 1,2,4,5-benzenetetraamine tetrahydrochloride than those that were not MYCN amplified (77). Notably, further studies with 1,2,4,5-benzenetetraamine tetrahydrochloride have shown no effect upon the viability of normal ganglion cells (Figure 4). These studies suggest that FAK is a potential therapeutic target for the treatment of human neuroblastoma, and may be most effective when directed toward aggressive, MYCN amplified tumors.
8. SUMMARY AND PERSPECTIVE
Neuroblastoma is a common childhood malignancy that continues to have a poor response to the current available therapies. It is clear that novel interventions will be required to treat this disease, especially in the advanced stages. Fairly good evidence has been generated to show that in neuroblastoma, focal adhesion kinase is correlated with MYCN oncogene amplification, and that inhibition of FAK in these amplified tumors results in decreased tumor cell survival. These findings underscore the potential for FAK inhibition in the development of new therapeutic strategies for neuroblastoma.
9. REFERENCES
1. R. Alex, O. Sozeri, S. Meyer, R. Dildrop: Determination of the DNA sequence recognized by the bHLH-zip domain of the N-myc protein. Nucleic Acids Res 20, 2257-2263 (1992)
doi:10.1093/nar/20.9.2257
2. G. Packham, J. L. Cleveland: C-myc and apoptosis. Biochim Biophys Acta 1242, 11-28 (1995)
3. D. M. Murphy, P. G. Buckley, K. Bryan, A. Das, L. Alcock, N. H. Foley, S. Prenter, I. Bray, K. M. Watters, D. Higgins, R. L. Stallings: Global MYCN transcription factor binding analysis in neuroblastoma reveals association with distinct E-box motifs and regions of DNA hypermethylation. PLoS One 4, e8154 (2009)
doi:10.1371/journal.pone.0008154
4. A. Negroni, S. Scarpa, A. Romeo, S. Ferrari, A. Modesti, G. Raschella: Decrease of proliferation rate and induction of differentiation by MYCN antisense DNA oligomer in a human neuroblastoma cell line. Cell Growth Diff 2, 511-518 (1991)
5. L. Whitesell, A. Rosolen, L. M. Neckers: Episome-generated N-myc antisense RNA restricts the differentiation potential of primitive neuroectodermal cell lines. Mol Cell Biol 11, 1360-1371 (1991)
6. P. J. Hurlin: N-myc functions in transcription and development. Birth Defects Res 75, 340-352 (2005)
doi:10.1002/bdrc.20059
7. S. Sawai, A. Shimono, Y. Wakamatsu, C. Palmes, K. Hanaoka, H. Kondoh: Defects of embryonic organogenesis resulting from targeted disruption of the N-myc gene in the mouse. Development 117, 1445-1455 (1993)
8. L. W. Stanton, M. Schwab, J. M. Bishop: Nucleotide sequence of the human N-myc gene. Proc Natl Acad Sci USA 83, 1772-1776 (1986)
doi:10.1073/pnas.83.6.1772
9. S. Adhikary, M. Eilers: Transcriptional regulation and transformation by MYC proteins. Nature 6, 635-645 (2005)
10. D. K. Thomas, C. A. Messam, B. A. Spengler, J. L. Biedler, R. A. Ross: Nestin is a potential mediator of malignancy in human neuroblastoma cells. J Biol Chem 279, 27994-27999 (2004)
doi:10.1074/jbc.M312663200
11. R. G. Harris, E. White, E. S. Phillips, K. A. Lillycrop: The expression of developmentally regulated proto-oncogene Pax-3 is modulated by N-myc. J Biol Chem 277, 34815-34825 (2002)
doi:10.1074/jbc.M109609200
12. C. F. Manohar, J. A. Bray, H. R. Salwen, J. Madafiglio, A. Cheng, C. Flemming, G. M. Marshall, M. D. Norris, M. Haber, S. L. Cohn: MYCN-mediated regulation of the MRP1 promoter in human neuroblastoma. Oncogene 23, 753-762 (2004)
doi:10.1038/sj.onc.1207151
13. A. B. West, G. Kapatos, C. O'Farrell, F. Gonzalez-de-Chavez, K. Chiu, M. J. Farrer, N. T. Maidment: N-myc regulates parkin expression. J Biol Chem 279, 28896-28902 (2004)
doi:10.1074/jbc.M400126200
14. G. Giannini, F. Cerignoli, M. Mellone, I. Massimi, C. Ambrosi, C. Rinaldi, C. Dominici, L. Frati, I. Screpanti, A. Gulino: High mobility group A1 is a molecular target for MYCN in human neuroblastoma. Cancer Res 65, 8308-8316 (2005)
doi:10.1158/0008-5472.CAN-05-0607
15. A. Porro, M. Haber, D. Diolaiti, N. Iraci, M. Henderson, S. Gherardi, E. Valli, M. A. Munoz, C. Xue, C. Flemming, M. Schwab, J. H. Wong, G. M. Marshall, G. Della Valle, M. D. Norris, G. Perini: Direct and coordinate regulation of ATP-binding cassette (ABC) transporter genes by MYC factors generates specific transcription signatures which significantly affect the chemoresistance phenotype of cancer cells. J Biol Chem Epub PMID: 20233711 (2010)
16. A. Dasgupta, S. K. Pierce, H. W. Findley: MycN is a transcriptional regulator of livin in neuroblastoma. Oncol Rep 22, 831-835 (2009)
17. J. Li, L. Kretzner: The growth-inhibitory Ndrg1 gene is a Myc negative target in human neuroblastomas and other cell types with overexpressed N- or c-myc. Molec Cell Biochem 250, 91-105 (2003)
doi:10.1023/A:1024918328162
18. E. Hatzi, C. Murphy, A. Zoephel, H. Ahorn, U. Tontsch, A. M. Bamberger, K. Yamauchi-Takihara, L. Schweigerer, T. Fotsis: N-myc oncogene overexpression down-regulates leukemia inhibitory factor in neuroblastoma. Oncogene 13, 803-812 (2002)
19. S. K. Oster, D. Y. L. Mao, J. Kennedy, L. Z. Penn: Functional analysis of the N-terminal domain of the Myc protein. Oncogene 22, 1998-2010 (2003)
doi:10.1038/sj.onc.1206228
20. G. M. Brodeur, R. C. Seeger, M. Schwab, H. E. Varmus, J. M. Bishop: Amplification of N-myc in untreated human neuroblastomas correlates with advanced stage disease. Science 224, 1121-1124 (1984)
doi:10.1126/science.6719137
21. L. Schweigerer, S. Breit, A. Wenzel, K. Tsunamoto, R. Ludwig, M. Schwab: Augmented MYCN expression advances the malignant phenotype of human neuroblastoma cells: evidence for induction of autocrine growth factor activity. Cancer Res 15, 4411-4416 (1990)
22. N. Gross, G. Miescher, D. Beck, S. Favre, C. Beretta: Altered growth and phenotype in clonal mycN transfectants of the SK-N-SH neuroblastoma cell line. Int J Cancer 59, 141-148 (1994)
doi:10.1002/ijc.2910590124
23. M. L. Schmidt, H. R. Salwen, C. F. Manohar, N. Ikegaki, S. L. Cohn: The biological effects of antisense N-myc expression in human neuroblastoma. Cell Growth Differ 5, 171-178 (1994)
24. K. Nara, T. Kusafuka, A. Yoneda, T. Oue, S. Sangkhathat, M. Fukuzawa: Silencing of MYCN by RNA interference induces growth inhibition, apoptotic activity, and cell differentiation in a neuroblastoma cell line with MYCN amplification. Int J Oncol 30, 1189-1196 (2007)
25. C. W. Woo, F. Tan, H. Cassano, J. H. Lee, K. C. Lee, C. J. Thiele: Use of RNA interference to elucidate the effect of MYCN on cell cycle in neuroblastoma. Pediatr Blood Cancer 50, 208-212 (2008)
doi:10.1002/pbc.21195
26. J. H. Kang, P. G. Rychahou, T. A. Ishola, J. Qiao, B. M. Evers, D. H. Chung: MYCN silencing induces differentiation and apoptosis in human neuroblastoma cells. Biochem Biophys Res Commun 351, 192-197 (2006)
doi:10.1016/j.bbrc.2006.10.020
27. W. A. Weiss, K. Aldape, G. Mohapatra, B. G. Feuerstein, J. M. Bishop: Targeted expression of MYCN causes neuroblastoma in transgenic mice. EMBO J 16, 2985-2995 (1997)
doi:10.1093/emboj/16.11.2985
28. H. C. Moore, K. M. Wood, M. S. Jackson, M. A. Lastowska, D. Hall, H. Imrie, C. P. Redfern, P. E. Lovat, F. Ponthan, K. O'Toole, J. Lunec, D. A. Tweddle: Histological profile of tumours from MYCN transgenic mice. J Clin Pathol 61, 1098-1103 (2008)
doi:10.1136/jcp.2007.054627
29. L. H. Xu, X. H. Yang, C. A. Bradham, D. A. Brenner, A. S. Baldwin, R. J. Craven, W. G. Cance: The focal adhesion kinase suppresses transformation-associated, anchorage-independent apoptosis in human breast cancer cells. Involvement of death receptor-related signaling pathways. J Biol Chem 275, 30597-30604 (2000)
doi:10.1074/jbc.M910027199
30. L. H. Xu, X.H. Yang, R. J. Craven, W. G. Cance: The COOH-terminal domain of the focal adhesion kinase induces loss of adhesion and cell death in human tumor cells. Cell Growth Differ 9, 999-1005 (1998)
31. V. A. Golubovskaya, S. Gross, A. S. Kaur, R. I. Wilson, L. H. Xu, X. H. Yang, W. G. Cance: Focal adhesion kinase N-terminus in breast carcinoma cells induces rounding, detachment and apoptosis. Mol Cancer Res 1, 755-764 (2003)
32. T. M. Weiner, E. T. Liu, R. J. Craven, W. G. Cance: Expression of focal adhesion kinase gene and invasive cancer. Lancet 342, 1024-1025 (1993)
doi:10.1016/0140-6736(93)92881-S
33. T. M. Weiner, E. T. Liu, R. J. Craven, W. G. Cance: Expression of growth factor receptors, the focal adhesion kinase, and other tyrosine kinases in human soft tissue tumors. Ann Surg Oncol 1, 18-27 (1994)
doi:10.1007/BF02303537
34. L. V. Owens, L. Xu, R. J. Craven, G. A. Dent, T. M. Weiner, L. Kornberg, E. T. Liu, W. G. Cance: Overexpression of the focal adhesion kinase (p125FAK) in invasive human tumors. Cancer Res 55, 2752-2755 (1995)
35. L. V. Owens, L. Xu, G. A. Dent, X. Yang, G. C. Sturge, R. J. Craven, W. G. Cance: Focal adhesion kinase as a marker of invasive potential in differentiated human thyroid cancer. Ann Surg Oncol 3, 100-105 (1996)
doi:10.1007/BF02409059
36. W. G. Cance, J. E. Harris, M. V. Iacocca, E. Roche, X. Yang, J. Chang, S. Simkins, L. Xu: Immunohistochemical analyses of focal adhesion kinase expression in benign and malignant human breast and colon tissues: correlation with preinvasive and invasive phenotypes. Clin Cancer Res 6, 2417-2423 (2000)
37. J. S. Chen, X. H. Huan, Q.Wang, X. L. Chen, X. H. Fu, X. H. Tan, L. J. Zhang, W. Li, J. Bi: FAK is involved in invasion and metastasis of hepatocellular carcinoma. Clin Exp Metastasis 27, 71-82 (2010)
doi:10.1007/s10585-010-9306-3
38. A. L. Lark, C. A. Livasy, B. Calvo, L. Caskey, D. T. Moore, X. Yang, W. G. Cance: Overexpression of focal adhesion kinase in primary colorectal carcinomas and colorectal liver metastases: immunohistochemistry and real-time PCR analyses. Clin Cancer Res 9, 215-222 (2003)
39. M. D. Schaller, C. A. Borgman, B. S. Cobb, R. R. Vines, A. B. Reynolds, J. T. Parsons: pp125FAK a structurally distinctive protein-tyrosine kinase associated with focal adhesions. Proc Natl Acad Sci USA 89, 5192-5196 (1992)
doi:10.1073/pnas.89.11.5192
40. S. K. Hanks, T. R. Polte: Signaling through focal adhesion kinase. BioEssays 19, 137-145 (1997)
doi:10.1002/bies.950190208
41. I. Zachary: Focal adhesion kinase. Int J Biochem Cell Biol 29, 929-934 (1997)
doi:10.1016/S1357-2725(97)00008-3
42. V. Gabarra-Niecko, M. D. Schaller, J. M. Dunty: FAK regulates biological processes important for the pathogenesis of cancer. Cancer Metastasis Rev 22, 359-374 (2003)
doi:10.1023/A:1023725029589
43. E. K. Han, T. Mcgonigal, J. Wang, V. L. Giranda, Y. Luo: Functional analysis of focal adhesion kinase (FAK) reduction by small inhibitory RNAs. Anticancer Res 24, 3899-3905 (2004)
44. C. A. Lipinski, N. L. Tran, E. Menashi, C. Rohl, J. Kloss, R. C. Bay, M. E. Berens, J. C. Loftus: The tyrosine kinase pyk2 promotes migration and invasion of glioma cells. Neoplasia 7, 435-445 (2005)
doi:10.1593/neo.04712
45. M. M. Shahzad, C. Lu, J. W. Lee, R. L. Stone, R. Mitra, L. S. Mangala, Y. Lu, K. A. Baggerly, C. G. Danes, A. M. Nick, J. Halder, H. S. Kim, P. Vivas-Mejia, A. N. Landen, G. Lopez-Berestein, R. L. Coleman, A. K. Sood: Dual targeting of EphA2 and FAK in ovarian carcinoma. Cancer Biol Ther 8,1027-34 2009
46. K. Tsutsumi, T. Kasaoka, H. M. Park, H. Nishiyama, M. Nakajima, T. Honda: Tumor growth inhibition by synthetic and expressed siRNA targeting focal adhesion kinase. Int J Oncol 33, 215-224 (2008)
47. L. H. Xu, L. V. Owens, G. C. Sturge, X. Yang, E. T. Liu, R. J. Craven, W. G. Cance: Attenuation of the expression of the focal adhesion kinase induces apoptosis in tumor cells. Cell Growth Diff 7, 413-418 (1996)
48. V. M. Golubovskaya, L. Beviglia, L. H. Xu, H. S. Earp, R. J. Craven, W. G. Cance: Dual inhibition of focal adhesion kinase and epidermal growth factor receptor pathways cooperatively induces death receptor-mediated apoptosis in human breast cancer cells. J Biol Chem 277, 38978-38987 (2002)
doi:10.1074/jbc.M205002200
49. Y. Zaizen, S. Taniguchi, S. Suita: The role of cellular motility in the invasion of human neuroblastoma cells with or without N-myc amplification and expression. J Pediatr Surg 33, 1765-1770 (1998)
doi:10.1016/S0022-3468(98)90281-0
50. L. A. Goodman, B. C. Liu, C. J. Thiele, M.L. Schmidt, S. L. Cohn, J. M. Yamashiro, D. S. Pai, N. Ikegaki, R. K. Wada: Modulation of N-myc expression alters the invasiveness of neuroblastoma. Clin Exp Metastasis 15, 130-139 (1997)
doi:10.1023/A:1018448710006
51. L. Ma, J. Young, H. Prabhala, E. Pan, P. Mestdagh, D. Muth, J. Teruya-Feldstein, F. Reinhardt, T. T. Onder, S. Valastyan, F. Westermann, F. Speleman, J. Vandesompele, R. A. Weinberg: miR-9, a MYC/MYCN-activated microRNA, regulates E-cadherin and cancer metastasis. Nat Cell Biol 12, 247-256 (2010)
doi:10.1038/ncb2024
52. R. Akeson, R. Bernards: N-myc down regulates neural cell adhesion molecule expression in rat neuroblastoma. Mol Cell Biol 10, 2012-2016 (1990)
53. R. Judware, L. A. Culp: Over-expression of transfected N-myc oncogene in human SKNSH neuroblastoma cells down-regulates expression of beta 1 integrin subunit. Oncogene 11, 2599-2607 (1995)
54. R. Judware, L. A. Culp: N-myc over-expression downregulates alpha3beta1 integrin expression in human Saos-2 osteosarcoma cells. Clin Exp Metastasis 15, 228-238 (1997)
doi:10.1023/A:1018417330479
55. N. Tanaka, M. Fukuzawa: MYCN downregulates integrin alpha1 to promote invasion of human neuroblastoma cells. Int J Oncol 33, 815-821 (2008)
56. L. Wu, J. A. Bernard-Trifilo, Y. Lim, S. T. Lim, S. K. Mitra, S. Uryu, M. Chen, C. J. Pallen, N. K. V. Cheung, D. Mikolon, A. Mielgo, D. G. Stupack, D. D. Schlaepfer: Distinct FAK-Crc activation events promote α5β1 and α4β1 integrin-stimulated neuroblastoma cell motility. Oncogene 27, 1439-1448 (2008)
doi:10.1038/sj.onc.1210770
57. P. S. Leventhal, E. A. Shelden, B. Kim, E. L. Feldman: Tyrosine phosphorylation of paxillin and focal adhesion kinase during insulin-like growth factor-I-stimulated lamellipodial advance. J Biol Chem 272, 5214-5218 (1997)
doi:10.1074/jbc.272.8.5214
58. B. Kim, E. L. Feldman: Differential regulation of focal adhesion kinase and mitogen-activated protein kinase tyrosine phosphorylation during insulin-like growth factor-I-mediated cytoskeletal reorganization. J Neurochem 71, 1333-1336 (1998)
59. B. Kim, E. L. Feldman: Insulin-like growth factor I prevents mannitol-induced degradation of focal adhesion kinase and Akt. J Biol Chem 277, 27393-27400 (2002)
doi:10.1074/jbc.M201963200
60. B. Kim, C. M. van Golen, E. L. Feldman: Degradation and dephosphorylation of focal adhesion kinase during okadaic acid-induced apoptosis in human neuroblastoma cells. Neoplasia 5, 405-416 (2003)
61. E. A. Beierle, N. A. Massoll, J. Hartwich, E. V. Kurenova, V. M. Golubovskaya, W. G. Cance, P. McGrady, W. B. London: Focal adhesion kinase expression in human neuroblastoma: immunohistochemical and real-time PCR analyses. Clin Cancer Res 14, 3299-3305 (2008)
doi:10.1158/1078-0432.CCR-07-1511
62. V. M. Golubovskaya, A. Kaur, W. G. Cance: Cloning and characterization of the promoter region of human focal adhesion kinase gene: nuclear factor kappa B and p53 binding sites. Biochim Biophys Acta 1678, 111-125 (2004)
63. E. A. Beierle, A. Trujillo, A. Nagaram, E. V. Kurenova, R. Finch, X. Ma, J. Vella, W. G. Cance: N-myc regulates focal adhesion kinase expression in human neuroblastoma. J Biol Chem 282,12503-12516 (2007)
doi:10.1074/jbc.M701450200
64. M. B. Calalb, T. R. Polte, S. K. Hanks: Tyrosine phosphorylation of focal adhesion kinase at site in the catalytic domain regulates kinase activity: a role for Src family kinases. Mol Cell Biol 15, 954-963 (1995)
65. Y. Sonoda, Y. Matsumoto, M. Funakoshi, D. Yamamoto, S. K. Hanks, T. Kasahara: Anti-apoptotic role of focal adhesion kinase (FAK). Induction of inhibitor of apoptosis proteins and apoptosis suppression by the overexpression of Fak in a human leukemic cell line, HL60. J Biol Chem 275, 16309-16315 (2000)
doi:10.1074/jbc.275.21.16309
66. H. C. Chen, J. L. Guan: Association of focal adhesion kinase with its potential substrate phosphatidylinositol 3-kinase. Proc Natl Acad Sci USA 91, 10148-10152 (1994)
doi:10.1073/pnas.91.21.10148
67. S. K. Hanks, L. Ryzhova, N. Y. Shin, J. Brabek: Focal adhesion kinase signaling activities and their implications in the control of cell survival and motility. Front Biosci 8, d982-996 (2003)
doi:10.2741/1114
68. D. D. Schlaepfer, S. K. Mitra: Multiple connections link FAK to cell motility and invasion. Curr Opin Genet Dev 14, 92-101 (2004)
doi:10.1016/j.gde.2003.12.002
69. S. Fulda, W. Lutz, M. Schwab, K. M. Debatin: MycN sensitizes neuroblastoma cells for drug-induced apoptosis. Oncogene 18, 1479-1486 (1999)
doi:10.1038/sj.onc.1202435
70. E. A. Beierle, X. Ma, A. Trujillo, E. V. Kurenova, W. G. Cance, V. M Golubovskaya: Inhibition of focal adhesion kinase and src increases detachment and apoptosis in human neuroblastoma cell lines. Mol Carcinog 49, 224-234 (2010)
71. Q. Shi, A. B. Hjelmeland, S. T. Keir, L. Song, S. Wickman, D. Jackson, O. Ohmori, D. D. Bigner, H. S. Friedman, J. N. Rich: A novel low-molecular weight inhibitor of focal adhesion kinase,TAE226, inhibits glioma growth. Mol Carcinog 46, 488-496 (2007)
doi:10.1002/mc.20297
72. S. Hatakeyama, D. Tomioka, E. Kawahara, N. Matsuura, K. Masuya, T. Miyake, I. Umemura, T. Kanazawa, T. Honda, O. Ohmori: Anti-cancer activity of NVP-TAE226, a potent dual FAK/IGF-IR kinase inhibitor, against pancreatic cancer. J Clin Oncol 18S, 13162 (2006)
73. E. Kawahara, O. Ohmori, K. Nonomura, Y. Murakami, D. Tomioka, S. Niwa, T. Meyer, J. Mestan, T. Honda, S. Hatakeyama: NVPTAE226, a potent dual FAK/IGF-IR kinase inhibitor, prevents breast cancer metastasis in vivo. J Clin Oncol 18S, 13163 (2006)
74. E. A. Beierle, A. Trujillo, A. Nagaram, V. M. Golubovskaya, W. G. Cance, E. V. Kurenova: TAE226 inhibits human neuroblastoma cell survival. Cancer Invest 14, 3299-3305 (2008)
75. V. M. Golubovskaya, C. Nyberg, M. Zheng, F. Kweh, A. Magis, D. Ostrov, W. G. Cance: A small molecule inhibitor, 1,2,4,5-benzenetetraamine tetrahydrochloride, targeting the y397 site of focal adhesion kinase decreases tumor growth. J Med Chem 51, 7405-7416 (2008)
doi:10.1021/jm800483v
76. D. Zheng, V. Golubovskaya, E. Kurenova, C. Wood, N. A. Massoll, D. Ostrov, W. G. Cance, S. N. Hochwald: A novel strategy to inhibit FAK and IGF-1R decreases growth of pancreatic cancer xenografts. Mol Carcinog 49, 200-209 (2010)
77. E. A. Beierle, X. Ma, J. Stewart, C. Nyberg, A. Trujillo, W. G. Cance, V. M. Golubovskaya: Inhibition of focal adhesion kinase decreases tumor growth in human neuroblastoma. Cell Cycle 9, 1005-1015 (2010)
Abbreviations: FAK: focal adhesion kinase
Key Words: MYCN, neuroblastoma, FAK, pediatric, SHEP-21/N, SK-N-AS, SK-N-BE(2), Review
Send correspondence to: Elizabeth A. Beierle, University of Alabama, Birmingham, 1600 7th Ave. South, ACC Room 300, Birmingham, AL 35233, Tel: 205-939-9688, Fax: 205-975-4972, E-mail: elizabeth.beierle@chsys.org