Effect of focal adhesion kinase (FAK) downregulation with FAK antisense oligonucleotides and 5-fluorouracil on the viability of melanoma cell lines

Smith, Charles S.; Golubovskaya, Vita M.; Peck, Erin K.; Xu, Lihui; Monia, Brett P.; Yang, Xihui; Cance, William G.

doi:10.1097/00008390-200510000-00003

Cited by 54 publications

(39 citation statements)

References 33 publications

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“…There are previous studies suggesting that FAK is a potential target for therapy in human tumors. Attenuation of FAK in vitro with either a dominant-negative FAK protein (41)(42)(43) or FAK antisense oligonucleotides (44,45) results in cellular rounding, detachment, and significant apoptosis in human tumor cells. Further, down-regulation of FAK in nontransformed fibroblasts (44), normal mammary cells (43), or other normal cell types (46) that express FAK does not result in increased apoptosis in these normal cells.…”

Section: Discussionmentioning

confidence: 99%

Focal Adhesion Kinase Expression in Human Neuroblastoma: Immunohistochemical and Real-time PCR Analyses

Beierle

Massoll²,

Hartwich³

et al. 2008

Clinical Cancer Research

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Purpose: The focal adhesion kinase (FAK) is a nonreceptor protein tyrosine kinase important in signaling between cells and their extracellular matrix. Studies have shown that FAK expression is up-regulated in several human tumors and is related to tumor progression. We recently found an increase in p125 FAK expression in human neuroblastoma cells lines and wished to determine its expression in human neuroblastoma specimens and evaluate for a possible correlation between p125 FAK expression and known prognostic factors for neuroblastoma. We hypothesized that p125 FAK expression would be up-regulated in advanced human neuroblastomas. Experimental Design: Using immunohistochemical techniques with monoclonal antibody 4.47 specific for p125 FAK expression, we analyzed 70 formalin-fixed, paraffin-embedded human neuroblastoma specimens for p125 FAK staining. In addition, real-time PCR was used to determine the abundance of FAK mRNA in 17 matched human neuroblastoma mRNA specimens. Results: FAK staining was present in 51of the 70 tumor specimens (73%). Immunohistochemical staining of p125 FAK in the ganglion-type tumor cells correlated with advanced International Neuroblastoma Staging System tumor stages and FAK mRNA abundance. In addition, p125FAK staining was significantly increased in stage IV tumors with amplification of the N-MYC oncogene. Conclusions: These novel findings provide evidence that FAK is expressed by advanced-stage neuroblastoma and provide a rationale for targeting FAK in the treatment of this tumor.Focal adhesion kinase (FAK) is a 125-kDa, nonreceptor protein tyrosine kinase that was originally isolated from embryonic chick fibroblasts transformed by v-src (1). FAK is localized to focal adhesions or contact points between cells and their extracellular matrix and has been shown to have important functions in integrin signaling, cellular motility, and cellular apoptosis. Integrins bind to FAK through their h subunits, leading to the phosphorylation of FAK (2), thereby creating a high-affinity binding site for the Src family kinases (3 -5). The FAK-Src complex activates protein kinases that eventually result in the activation of nuclear transcription factors leading to cell differentiation and growth. In addition, phosphorylation of FAK results in the binding and activation of other signaling molecules (6) resulting in decreased apoptosis and increased cellular survival.FAK has been shown to be overexpressed in several human tumors (7), with numerous reports of significantly increased FAK protein expression in primary and metastatic breast cancer (8 -11) and in colon (8, 12, 13), thyroid (14, 15), ovarian (16), and esophageal cancers (17), head and neck tumors (18), and melanoma (19). The expression of FAK mRNA has also been shown to correlate with tumor aggressiveness, with an increased abundance of FAK mRNA being seen during the progression of epithelial tumors to metastatic phenotypes (20) and also in the hepatic metastasis of human colorectal carcinomas (12). Data correlating increased fak...

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Section: Discussionmentioning

confidence: 99%

Focal Adhesion Kinase Expression in Human Neuroblastoma: Immunohistochemical and Real-time PCR Analyses

Beierle

Massoll²,

Hartwich³

et al. 2008

Clinical Cancer Research

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“…N-MYC is reported to be amplified in human melanoma and sarcomas (62) and is associated with poor outcomes (63). It has also been shown that FAK is overexpressed in human sarcoma (16) and melanoma tumors (64,65) and that down-regulation of FAK results in decreased survival in human melanoma cells (66). The regulation of FAK expression by N-MYC in these tumor types has not been investigated but may contribute to the molecular regulation of FAK in these tumors.…”

Section: Discussionmentioning

confidence: 99%

N-MYC Regulates Focal Adhesion Kinase Expression in Human Neuroblastoma

Beierle

Trujillo

Nagaram

et al. 2007

Journal of Biological Chemistry

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N-MYC is a transcription factor important for the control of cellular differentiation and proliferation (1, 2). It is normally expressed in the brain and peripheral nervous system (1). N-MYC has been shown to influence the differentiation of neural crest cells. Early in mouse embryogenesis, N-MYC is present primarily in the migrating neural crest cells, but as the embryo matures, the expression of N-MYC becomes limited to those cells that are undergoing neuronal differentiation (3). Abnormal expression of N-MYC is most notably associated with the pediatric tumor of neural crest origin, neuroblastoma. Amplification of the N-MYC oncogene is the primary adverse prognostic indicator in human neuroblastoma (4, 5). The level of N-MYC expression has been shown to correlate with the growth (6 -9) and invasiveness (10) of neuroblastoma cells, and transgenic mice with N-MYC overexpression develop spontaneous neuroblastoma tumors (11). In addition, down-regulation of N-MYC with antisense oligonucleotides leads to decreases in both cellular proliferation and in anchorageindependent growth in the neuroblastoma cells (6, 9). Despite this information, the exact function and transcriptional gene targets of N-MYC in neuroblastoma are currently not well characterized (5).Focal adhesion kinase (FAK) 3 is a nonreceptor cytoplasmic 125-kDa protein-tyrosine kinase. Initial studies revealed that both the transcription of FAK mRNA (12) and the expression of FAK protein (13-19) are significantly increased in primary and metastatic breast, colon, and thyroid tumors compared with normal tissues 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 with control tissues (20), suggesting that the increased FAK expression in human tumors occurs at the level of transcription. Recently the FAK promoter was cloned and characterized, and transcriptional regulation of the FAK promoter has been demonstrated (21).FAK controls a number of cell signaling pathways including proliferation, viability, motility, and survival (22-25). The inhibition of FAK with antisense oligonucleotides has been shown to cause decreased growth in tumor cells (26). In addition, FAK inhibition with a dominant-negative FAK protein (FAK-CD) inhibited cell growth in human melanoma cells (12) and in * The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

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“…Treatment of A549 human adenocarcinoma cells with FAK antisense oligonucleotides significantly reduced FAK expression and the concomitant signaling to c-Jun NH 2 -terminal kinase; cell migration and invasion through Matrigel were also inhibited (41). Treatment of melanoma cell lines with FAK antisense oligonucleotides resulted in the decreased expression of FAK and a 2.5-fold increase in cell death compared with treatment with control oligonucleotides (42).…”

Section: Validation Of Fak As a Therapeutic Targetmentioning

confidence: 94%

Focal Adhesion Kinase: Targeting Adhesion Signaling Pathways for Therapeutic Intervention

Parsons

Slack‐Davis

Tilghman

et al. 2008

Clinical Cancer Research

108

118

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The tumor microenvironment plays a central role in cancer progression and metastasis.Within this environment, cancer cells respond to a host of signals including growth factors and chemotactic factors, as well as signals from adjacent cells, cells in the surrounding stroma, and signals from the extracellular matrix. Targeting the pathways that mediate many of these signals has been a major goal in the effort to develop therapeutics. Integrin Signals and CancerIntegrins constitute the primary set of receptors for extracellular matrix (ECM) components within the tumor microenvironment, and integrin expression is often modulated during cancer progression and metastasis (1 -6). Integrins are heterodimeric transmembrane receptors that link the cell to the ECM by direct binding to the actin cytoskeleton through a scaffold of several actin-binding proteins (7). Thus, they serve as sites of contact between the cell and the ECM. In adherent cells such as fibroblasts, these sites of contact are termed focal adhesions (8). Focal adhesions serve as nodes to transmit environmental signals initiated by soluble growth factors, ECM, and surrounding mechanical forces to regulate cell growth and migration (9 -11). Within tumors, cancer cells respond to changes in the expression and organization of matrix proteins such as fibronectin and collagen. In addition, tumor cells secrete proteases that cleave ECM molecules (metalloproteases) providing both growth-promoting remodeling of the tumor microenvironment and escape of tumor cells through the basement membrane (12).Tyrosine protein kinases play a key role in regulating signals mediated by integrin receptors (13,14). A central regulator of integrin signaling is focal adhesion kinase (FAK), a ubiquitously expressed nonreceptor tyrosine protein kinase (15). FAK expression is required for many normal cellular functions (16) and its expression is up-regulated in a variety of late-stage cancers (16 -19). Here we review the role of FAK in promoting integrin and growth factor signals and its link to cancer progression. We also discuss experiments validating FAK as a therapeutic target, summarize the current state of development of small-molecule inhibitors of FAK, and report progress in assessing such inhibitors in human clinical trials.FAK structure and regulation. FAK is recruited to focal adhesions and activated upon integrin engagement with the ECM (15, 16). The overall molecular structure of FAK is consistent with its function as a protein scaffold, in addition to its function as a tyrosine kinase. The central catalytic domain is flanked upon the NH 2 terminus by a FERM domain and COOH-terminal proline-rich and focal adhesion targeting domains (Fig. 1A). The FERM domain has been reported to serve as a site of interaction with a variety of molecules including integrin cytoplasmic domains, ezrin, Arp2/3, p53, and membrane receptors (13,15). The COOH-terminal domain functions as a scaffold with two proline-rich protein interaction domains (PR-I, PR-II) and a focal adhesion targe...

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Effect of focal adhesion kinase (FAK) downregulation with FAK antisense oligonucleotides and 5-fluorouracil on the viability of melanoma cell lines

Cited by 54 publications

References 33 publications

Focal Adhesion Kinase Expression in Human Neuroblastoma: Immunohistochemical and Real-time PCR Analyses

Focal Adhesion Kinase Expression in Human Neuroblastoma: Immunohistochemical and Real-time PCR Analyses

N-MYC Regulates Focal Adhesion Kinase Expression in Human Neuroblastoma

Focal Adhesion Kinase: Targeting Adhesion Signaling Pathways for Therapeutic Intervention

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