Tyrosine phosphorylation as a marker for aberrantly regulated growth‐promoting pathways in cell lines derived from head and neck malignancies

Cardinali, M.; Pietraszkiewicz, Halina; Ensley, John F.; Robbins, K C

doi:10.1002/ijc.2910610117

Cited by 128 publications

(96 citation statements)

References 18 publications

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“…For this analysis, we selected a panel of 11 HNSCC cell lines derived from primary and secondary cancer lesions of contrasting clinical staging (11). These cells have been comprehensively explored for alterations of major tumor suppressor genes (15,16).…”

Section: Resultsmentioning

confidence: 99%

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Persistent Activation of the Akt Pathway in Head and Neck Squamous Cell Carcinoma

Amornphimoltham

Sriuranpong

Patel

et al. 2004

Clinical Cancer Research

165

143

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Squamous carcinomas of the head and neck (HNSCC) represent the sixth most common cancer among men worldwide and a major cause of morbidity and mortality due to its relatively poor prognosis. As part of ongoing studies addressing the molecular events underlying tumor progression in HNSCC, we have explored the nature of the proliferative pathways in which dysregulation may promote aberrant cell growth in this tumor type. The serine/threonine protein kinase Akt is a downstream target of phosphatidylinositol 3-kinase and a key regulator of normal and cancerous growth and cell fate decisions. Therefore, in this study, we have examined the status of activation of Akt in different stages of squamous cell carcinoma development in mice and in clinical samples from HNSCC patients. By immunohistochemical analysis, using a recently developed phosphorylation state-specific antibody, we demonstrated that Akt activation correlates closely with the progression of mouse skin squamous cell carcinoma. We also observed that activation of Akt is a frequent event in human HNSCC because active Akt can be detected in these tumors with a pattern of expression and localization correlating with the progression of the lesions. In line with these observations, Akt was constitutively activated in a large fraction of HNSCCderived cell lines. We also provide evidence that the Akt signaling pathway may represent a biologically relevant target for a novel antineoplastic agent, UCN-01, which recently has been shown to be active in cellular and xenograft models for HNSCC at concentrations safely achievable in clinically relevant situations.

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Section: Resultsmentioning

confidence: 99%

“…HNSCC cell lines HN4, HN6, HN8, HN12, HN13, HN17, HN19, HN22, HN26, HN30, and HN31 and HaCaT cells were maintained in DMEM with 10% FCS, penicillin, and streptomycin in 5% CO 2 at 37°C as described previously (11).…”

Section: Methodsmentioning

confidence: 99%

Persistent Activation of the Akt Pathway in Head and Neck Squamous Cell Carcinoma

Amornphimoltham

Sriuranpong

Patel

et al. 2004

Clinical Cancer Research

165

143

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“…Materials were obtained from Sigma-Aldrich, unless otherwise indicated. WSU-HN13 cells (5) were transfected using Polyfect (Qiagen) with a plasmid (pCEFL2) driving expression of TVA, the receptor for RCAS viruses, from the EF1a promoter and selected for resistance to 0.4 mg/mL G418. Clones were screened for TVA by fluorescence after immunostaining with antibodies and transduction with enhanced green fluorescent protein (EGFP)-Renilla luciferase (RLh) cloned into RCASYGWB, an RCAS proviral vector (6).…”

Section: Methodsmentioning

confidence: 99%

Snail Up-regulates Proinflammatory Mediators and Inhibits Differentiation in Oral Keratinocytes

et al. 2008

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The transcriptional repressor Snail2 is overexpressed in head and neck squamous cell carcinomas (HNSCC) relative to nonmalignant head and neck mucosal epithelium, and in locally recurrent relative to nonrecurrent HNSCCs. We investigated the mechanisms by which Snails might contribute to the pathogenesis of HNSCCs using cell biological and molecular analyses. Oral keratinocytes that expressed Snails acquired an enhanced ability to attract monocytes and to invade a dense interstitial collagen matrix. They were also found to up-regulate production of proinflammatory cytokines and cyclooxygenase-2 (COX2), which have previously been shown to correlate with malignancy. Induction of nuclear factor-KB transcriptional activity by Snails was weak and not sufficient to account for the elevated levels of COX2, interleukin (IL)-6, IL8, or CXCL1. In addition, expression of Snails in oral keratinocytes impaired desquamation in vitro and strongly repressed expression of both ELF3 and matriptase-1, which play important roles in the terminal differentiation of keratinocytes. Reexpression of matriptase-1 in Snail-expressing cells partially rescued desquamation. This implicates Snails as contributing to malignancy both at the early stages, by impeding terminal differentiation, and at later stages, when invasion and inflammation are important.

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“…Human epithelial cell lines SCC25, 26 T45, 27 BICR16, 28 HN6, HN12, HN13, HN26 and HN30, 29 derived from HNSCC, and DOK, derived from a premalignant oral lesion, 30 were cultured in DMEM supplemented with 10% FBS and 0.4 g/ml hydrocortisone on a feeder layer of lethally irradiated Swiss-3T3 fibroblasts. Feeder cells were removed prior to subculturing or RNA preparation by standard procedures.…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

Laminin‐γ2 overexpression in head‐and‐neck squamous cell carcinoma

Patel

Aldridge

Ensley

et al. 2002

Intl Journal of Cancer

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To identify molecular markers for the progression of headand-neck squamous cell carcinoma (HNSCC), we used RNA arbitrarily primed (RAP) PCR to determine the qualitative and quantitative differences in gene expression between normal epithelial cells, those derived from dysplastic oral mucosa and invasive and metastatic HNSCC. Three differentially expressed DNA fragments (RAP20, RAP21, RAP26) that were upregulated in a tumor cell line (T45) were identified as being regions of the ␥2 subunit of human laminin-5. Northern blot analysis of total cellular RNA revealed overexpression of these transcripts in 6 of 7 HNSCC cell lines compared with normal epidermal keratinocytes. In contrast, no differences were observed in HeLa (cervical carcinoma) or HCT116 (colon carcinoma) cells. Laminin is a major component of the basement membrane, which performs multiple biologic roles, including cell attachment, spreading and migration as well as being involved in cellular proliferation and differentiation. 1,2 Structurally, laminin is a heterotrimeric protein comprising ␣, ␤ and ␥ subunits, of which several variants exist. 3 Thus, different subunit combinations result in a family of laminins. 3,4 Specific biologic activities of different laminins may be reflected in the fact that these molecules show tissue-specific expression. For example, laminin-5 is considered to be an epithelial laminin as it is found as a component of basement membranes in epithelial tissues but not in mesenchyme. 5,6 Furthermore, cytokines that regulate epithelial homeostasis, such as transforming growth factor (TGF)-␤, modulate the expression of laminin-5 polypeptides in epidermal keratinocytes. 7 A role for laminin isoforms in tumor cell invasion is becoming apparent. 8 For instance, attachment of cancer cells is mediated in part by laminins. 9,10 Additionally, increased cell-surface expression of laminin was found in highly metastatic fibrosarcoma cells compared to cells of low metastatic potential. 11 Furthermore, expression and activity of matrix metalloproteases, which play critical roles in cellular invasion, are elevated in the presence of laminin, 12 and metastasis of melanoma cells is also enhanced. 13 Further studies have demonstrated that attachment and metastasis of tumor cells can be inhibited by incubation with antilaminin antibodies 14,15 or synthetic laminin peptides. 16 Several reports have implicated laminin-5 in epithelial tumor cell invasion. In one study, overexpression of the ␥2 chain of laminin-5 was found in 26 of 27 carcinomas, while various mesenchymal tumors did not express this molecule. 17 Furthermore, the highest expression of ␥2 occurred at areas of active invasion in colon adenocarcinomas, and a clear correlation was found with tumor budding. 17 In a subsequent study, these findings were extended to examine expression in a larger series of human cancers, 18 demonstrating that levels of ␥2 were indeed the highest in squamous carcinomas, particularly in cells adjacent to surrounding stromal tissue and in deeply invading cell ne...

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Tyrosine phosphorylation as a marker for aberrantly regulated growth‐promoting pathways in cell lines derived from head and neck malignancies

Cited by 128 publications

References 18 publications

Persistent Activation of the Akt Pathway in Head and Neck Squamous Cell Carcinoma

Persistent Activation of the Akt Pathway in Head and Neck Squamous Cell Carcinoma

Snail Up-regulates Proinflammatory Mediators and Inhibits Differentiation in Oral Keratinocytes

Laminin‐γ2 overexpression in head‐and‐neck squamous cell carcinoma

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