This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Rodrigo, J. P. Articles by Suárez, C. Search for Related Content PubMed PubMed Citation Articles by Rodrigo, J. P. Articles by Suárez, C.

EMS1 Gene Amplification Correlates with Poor Prognosis in Squamous Cell Carcinomas of the Head and Neck¹

Departments of Otolaryngology-Head and Neck Surgery [J. P. R., L. A. G., C. S.] and Molecular Biology [S. R., P. S. L.], Hospital Central de Asturias, University of Oviedo, Instituto Universitario de Oncología, 33006 Oviedo, Spain

	ABSTRACT

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
The relationship between CCND1 and/or EMS1 amplification and disease outcome was studied in a prospective series of 104 head and neck squamous cell carcinomas treated by surgical resection. The CCND1 and EMS1 copy number in tumor samples was estimated by differential PCR. The presence or absence of amplification was analyzed in relation to clinicopathological variables, tumor recurrence, and patient survival. CCND1 amplification occurred in 32 cases (31%) and was associated with increased lymph node stage (P = 0.005) and advanced disease stage (P = 0.003). EMS1 amplification was identified in 21 cases (20%) and was related with advanced T stages (P = 0.001), increased lymph node stage (P = 0.02), advanced disease stage (P = 0.041), poor histological differentiation (P = 0.018), recurrent disease (P = 0.0004), and reduced disease-specific survival (P < 0.0001). Coamplification of both genes occurred in 11 cases (11.5%). Multivariate analysis confirmed that in addition to regional lymph node status, EMS1 amplification is an independent predictor of death from the tumor (P = 0.0027). CCND1 amplification was not prognostic. These data indicate that EMS1 amplification, but not CCND1 amplification, predicts early recurrence and reduced survival in squamous cell carcinoma of the head and neck. The prognostic significance previously attributed to CCND1 amplification may be attributable to its frequent coamplification with EMS1.

	INTRODUCTION

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
The survival rate for patients with SCCHN³ (1) has remained unchanged in recent years despite the advances in diagnosis and treatment. Our ability to prognosticate for advanced cases of SCCHN is particularly poor owing to variations in the biological behavior of the tumors and inadequacies of the present staging system. It is therefore essential to identify new markers (e.g., genetic markers) that may distinguish differences in tumor behavior.

Multiple genes and chromosomal regions are implicated in SCCHN tumorigenesis. Amplification of chromosomal region 11q13 is one of the genetic alterations most frequently observed in SCCHN (1, 2, 3, 4, 5, 6, 7) . The amplified 11q13 region is estimated to be 3–5 megabases in size and includes four putative oncogenes: CCND1 (PRAD1), FGF3 (INT2), FGF4 (HST1), and EMS1. CCND1 and EMS1 are the more prominent candidate oncogenes because they were found to be also overexpressed in all carcinomas with an 11q13 amplification (8) . Therefore, the activation of these genes might confer the selective advantage to these tumors. The CCND1 gene encodes the cell cycle regulating cyclin D1 protein involved in the G₁-to-S transition. EMS1 encodes a cytoskeletal protein homologous to the avian F actin-binding protein (cortactin), which is thought to be involved in cell-to-cell interactions (8) .

Amplification of the chromosome 11q13 region in SCCHN has been correlated with aggressive tumor growth (3 , 9) , the presence of lymph node metastases (7 , 10, 11, 12) , and poor prognosis (9) . In addition, various studies have assessed the clinical and prognostic significance of CCND1 amplification and/or overexpression, showing an association with recurrence and shortened overall survival (6 , 13, 14, 15, 16, 17) . However, little is known about the prognostic significance of EMS1 amplification in SCCHN. Because EMS1 amplification in tumors results in overexpression of cortactin and this is accompanied by a partial redistribution of cortactin from the cytoplasm into cell-matrix contact sites, it has been suggested that EMS1 amplification can contribute to the invasive potential of tumor cells (8 , 18) .

We hypothesized that, although both CCND1 and EMS1 contribute to the growth advantage of SCCHN with 11q13 amplification, amplification of each gene individually may confer different properties to these tumors. The purpose of this study was therefore to assess the correlation between CCND1 and/or EMS1 amplification, the clinicopathological characteristics, and tumor behavior in SCCHN.

To evaluate CCND1 and EMS1 amplifications, we have used differential PCR, which is a rapid, simple, and sensitive method to determine semiquantitatively the number of gene copies. It only requires small amounts of DNA, which facilitates these studies in clinical medicine.

	MATERIALS AND METHODS

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
Tumor Specimens.
Tissue samples of 104 consecutive primary SCCHN were obtained from patients undergoing surgical resection of their tumor between January 1992 and July 1994. All patients included in our study had a single primary tumor, none had undergone treatment before surgery, and had microscopically clear surgical margins. None of the patients were thought to have had distant metastases at the time of surgery. All but one subject were smokers, and 94 of them were also habitual alcohol drinkers. The exceptionally low proportion of women in the sample (only one) is usual for this kind of tumor in our media. A total of 56 patients received postoperative radiotherapy. As a general rule, this was administered to the patients with histologically N₂ or N₃ neck lesions, and also to the patients with N₀-N₁ neck lesions with locally advanced stage (T₄). The clinicopathological data from the patients are shown in Table 1 . The stage of disease was determined after the surgical resection of the tumor (with lymph node dissection where appropriate) according to the TNM system of the Union Internationale Contre le Cancer (Ed. 4).

The PCR mixtures contained 0.2–0.5 µg of target DNA, 10 mM Tris-HCl (pH 8.3), 1.5 mM MgCl₂, 50 mM KCl, 0.2 mM each of deoxynucleotide triphosphate, 1 µM of each primer, and 1 unit of Taq polymerase (Boehringer Mannheim, Mannheim, Germany) in a total volume of 50 µl with a 50-µl mineral oil overlay. The PCR cycles included 1 min at each temperature (94°C, 56°C, and 72°C), for a total of 30 cycles, and a final cycle at 72°C for 7 min. Two different sets of primers, one for the target gene (CCND1or EMS1) and the other for the control gene, were present simultaneously in the reaction vessel, as previously described (20 , 21) . The TH gene, which is located on the same chromosome as the target genes, was used as the control gene. Primer sequences were designed from the genomic sequences (obtained from GenBank).⁴ Primers for the CCND1 gene were 5'-CGTACCCCG-ATGCCAACC and 5'-ATGGACGGCAGGACCTCC, and they amplified a fragment of 121bp. Primers for the EMS1 gene were 5'-TCCCCTGATGCCCAGGTC and 5'-TCC-CAATCCAGAGACCCG, and they amplified a sequence of 111 bp. Primers for the TH gene were 5'-GCCCCAGCTGCATCCTAC and 5'-CTTGGCAGACACCTGGGG, and they amplified a sequence of 188 bp. The primers were purchased from MWG-Biotech (Mannheim, Germany).

Samples of DNA from normal tissues (tonsils) obtained from noncancer patients were used as negative controls. As positive controls, a mixture of DNA from normal tissue and increasing amounts of a previously PCR-amplified sequence of the target gene (CCND1 or EMS1), mimicking different degrees of amplification, was used as template in the PCR reaction, as previously described (19) .

Electrophoresis and Quantitation of PCR Products.
After PCR, 10 µl of each sample were electrophoresed on gels containing 3% NuSieve agarose (FMC, Rockland, ME) and 1% molecular biology grade agarose (Promega, Madison, WI) for 1.5 h at 65 V in 40 mM Tris-Acetate and 2 mM EDTA buffer. The gels were stained with ethidium bromide, and the images of the UV-illuminated gels were captured using a digital camera and stored in an IBM-compatible PC system. The bands were thereafter quantitated by computerized densitometric analysis techniques (Kodak Digital Science 10, Eastman Software, Billerica, MA), and the CCND1 or EMS1:TH ratios were determined. The results of densitometry were corroborated by visual inspection of the gels.

Statistical Analysis.
Statistical analysis was performed using ², with Yates’ correction where appropriate, and Fisher’s exact tests. Survival curves were calculated using the Kaplan-Meier product limit estimate (22) . Deaths from causes other than the index tumor or its metastases were not considered treatment failures, and these patients were censored in all analysis involving the length of survival. Differences between survival times were analyzed by the log-rank method (23) . Multivariate Cox proportional hazards models (24) were used to examine the relative impact of either variables demonstrated to be statistically significant in univariate analysis or those variables likely to have an effect on outcome (e.g., tumor site and histopathological grade). In these models, tumor sites were dichotomized as oral cavity and pharynx versus larynx. Similarly, T-stage classification was dichotomized as T₁, T₂, or T₃ versus T₄. Lymph node metastasis classification was dichotomized as N₀ or N₁ versus N₂ or N₃. Stage was dichotomized as stage I, stage II, or stage III versus stage IV. Finally, histopathological grade was dichotomized as well- and moderately differentiated versus poorly differentiated. Ps 0.05 were considered to be statistically significant. Patients were observed for at least 36 months.

	RESULTS

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 
Differential PCR.
A total of 104 samples from patients with SCCHN were studied. Each sample was investigated for CCND1 and EMS1 amplifications by differential PCR using the gene for TH as the control. Samples of DNA obtained from normal tissues showed CCND1:TH ratios of 1.2 ± 0.21 (mean ± SD) and EMS1:TH ratios of 0.9 ± 0.1 (mean ± SDs obtained from 10 different samples). The positive controls showed that there was an increase in the PCR product from the target gene, compared with the PCR product from the control gene, as the target gene copy number used as the template was increased (data not shown). Experimentally, gene amplification was defined as at least a 2-fold increase in the CCND1:TH or EMS1:TH ratios, relative to the ratios obtained with normal tissue DNA. Because of the semiquantitative nature of the differential PCR, the ratios obtained were not converted to a score of amplification; the results were expressed as amplification or no amplification of the CCND1 or EMS1 genes. PCR was carried out at least twice in the positive cases.

CCND1 and EMS1 amplifications were found in 32 (31%) and 21 (20%) of the 104 tumor samples, respectively. Coamplification of both genes was demonstrated in 11 cases (11.5%). None of the cases considered nonamplified presented a CCND1:TH ratio higher than 1.9 (mean ± SD, 1.3 ± 0.2), or an EMS1:TH ratio higher than 1.8 (mean ± SD, 1.2 ± 0.2). Fig. 1 shows the results of the differential PCR in some of the amplified cases.

View larger version (38K): [in this window] [in a new window]   Fig. 1. Differential PCR analysis of CCND1 (A) and EMS1 (B) gene amplifications from various tumor samples in which gene amplification was found. The PCR products were separated by agarose gel electrophoresis and quantified by image analysis densitometry. N, controls with normal tissue DNA; C+, the positive controls (2-fold amplification); C-, the negative controls.

View larger version (17K): [in this window] [in a new window]   Fig. 2. Kaplan-Meier disease-free survival curves of the patients with (----) and without () CCND1 (A) or EMS1 (B) gene amplification. Differences between survival times were analyzed by the log-rank method.

	DISCUSSION

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

In SCCHN, amplification of 11q13 has been reported in 20–52% of patients (1, 2, 3 , 5 , 7 , 9, 10, 11) , which compares well with 34% in the present study. Several works have focused on CCND1 amplification and showed similar results (4 , 6 , 13 , 17) . In contrast, only a few studies have also analyzed EMS1 amplification (3 , 8 , 9 , 12) . In these series, as in our study, CCND1 and EMS1 have not been uniformly amplified in all cases. These differences in amplification of 11q13 markers were also described in breast carcinomas (27) , and they are in agreement with the structure of the 11q13 amplicon proposed by Schuuring (8) : the amplified 11q13 region harbors at least three, or even more, separate cores of amplification. The first core extends from BCL1 to FGF4, with the CCND1 gene as the best candidate key gene. The second core extends from FGF3 to the EMS1 gene, with the EMS1 gene as the best candidate key gene. Another two cores of amplification have been described in the centromeric and the telomeric borders, respectively, of the 11q13 region; as yet, no gene has been identified in these subregions.

The results of the present study indicate that the amplification of EMS1 is of prognostic value for SCCHN independent of other known risk factors. In fact, all of the patients with EMS1 amplification experienced disease recurrence. In contrast, CCND1 amplification has not found to be of prognostic significance in multivariate analysis. In other reports, 11q13 amplification (without reference to specific genes) was observed in patients with advanced disease, a poorly differentiated histology of the tumor, and deeply invasive growth (3) . In concordance with the presumed association with progressed disease, the amplified cases develop more frequent recurrences and have an increased risk of tumor-associated death (6 , 7 , 9) . Opposite reports (5 , 11) indicate the controversy on this point. This may be attributable to the use of different 11q13 markers, measuring only one of the amplification cores to analyze 11q13 amplification. On the other hand, the amplification and/or overexpression of CCND1 has been reported to be a poor prognostic sign in various studies focused on this gene (6 , 13, 14, 15, 16, 17) . Again, there are also reports on the contrary (4 , 28) . In addition, the association of CCND1 amplification and/or overexpression and poor prognosis is not absolute, suggesting that amplification and/or overexpression of CCND1 contributes only partially to the process of tumor progression. Our findings indicate that the amplification of CCND1 loses its prognostic significance when the cases in which CCND1 is coamplified with the EMS1 gene are eliminated. Thus, the prognostic significance attributed to the CCND1 amplification may be attributable to its frequent coamplification with the EMS1 gene. This agrees with the proposed role of these genes in tumor development (8) : CCND1 overexpression might confer to the tumor cells the ability to proliferate under reduced growth factor conditions because of its effects in the G₁-to-S transition of the cell cycle; EMS1 overexpression might mediate the increased invasive and metastatic behavior of tumor cells attributable to its effects in the organization and the functioning of cytoskeleton and cell adhesion structures.

Differential PCR has previously been shown as a simple, rapid, and sensitive method to detect genetic amplification (17 , 19, 20, 21) . Our results corroborate that of others, showing that this method is suitable to detect gene amplification. It is reproducible, and its main advantage is that small quantities of DNA are needed, making possible its clinical use with samples obtained from small biopsies or fine needle aspirations. Using this technique, we have found an 11q13 amplification rate in agreement with previous studies that used conventional techniques.

We conclude that amplification of the EMS1 gene is an independent prognostic marker for survival, and it specifies a subgroup of operable SCCHN patients that is at increased risk and might benefit from more intensive treatment and follow-up. Whether this is the result of greater tumor aggressiveness or of decreased responsiveness to adjuvant radiotherapy remains to be elucidated. Presently, clinical and histopathological parameters are used as guides for the application of adjunctive therapy, such as radiotherapy or even resection of the surgical field. Our data suggest that EMS1 amplification can augment the predictive power of those clinical and histopathological markers. Routine implementation of this molecular test will require confirmation of our findings in larger prospective studies.

	ACKNOWLEDGMENTS

 
We thank Dr. Enrique F. Bustillo for assistance with statistical analysis.

	FOOTNOTES

 
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.

¹ Supported by Grant 97/1092 from the Fondo de Investigaciones Sanitarias, Spain.

² To whom requests for reprints should be addressed, at Servicio de Otorrinolaringología, Hospital Central de Asturias, 33006 Oviedo, Spain. Phone: 34-985112109; E-mail: jrodrigot{at}seorl.org

³ The abbreviations used are: SCCHN, squamous cell carcinoma of the head and neck; TH, tyrosine hydroxylase.

⁴ Internet address: http://www.ncbi.nlm.nih.gov.

Received 1/24/00; revised 5/ 3/00; accepted 5/ 3/00.

	REFERENCES

Top ABSTRACT INTRODUCTION MATERIALS AND METHODS RESULTS DISCUSSION REFERENCES

 

1. Merritt W. D., Weissler M. C., Turk B. F., Gilmer T. M. Oncogene amplification in squamous cell carcinoma of the head and neck. Arch. Otolaryngol. Head Neck Surg., 116: 1394-1398, 1990.
2. Leonard J. H., Kearsley J. H., Chenevix-Trench G. E., Hayward N. K. Analysis of gene amplification in head and neck squamous cell carcinomas. Int. J. Cancer, 48: 511-515, 1991.[Medline]
3. Williams M. E., Gaffey M. J., Weiss L. M., Wilczynski S. P., Schuuring E., Levine P. A. Chromosome 11q13 amplification in head and neck squamous cell carcinoma. Arch. Otolaryngol. Head Neck Surg., 119: 1238-1243, 1993.
4. Callender T., El-Naggar A. K., Lee M. S., Frankenthaler R., Luna M. A., Batsakis J. G. PRAD-1 (CCND1)/cyclin D1 oncogene amplification in primary head and neck squamous cell carcinoma. Cancer (Phila.), 74: 152-158, 1994.[CrossRef][Medline]
5. Fortin A., Guerry M., Guerry R., Talbot M., Parise O., Schwaab G., Bosq J., Bourhis J., Salvatori P., Janot F., Busson P. Chromosome 11q13 gene amplifications in oral and oropharyngeal carcinomas: no correlation with subclinical lymph node invasion and disease recurrence. Clin. Cancer Res., 3: 1609-1614, 1997.[Abstract]
6. Akervall J. A., Michalides R. J. A. M., Mineta H., Balm A., Borg A., Dictor M. R., Jin Y., Loftus B., Mertens F., Wennerberg J. P. Amplification of cyclin D1 in squamous cell carcinoma of the head and neck and the prognostic value of chromosomal abnormalities and cyclin D1 overexpression. Cancer (Phila.), 79: 380-389, 1997.[CrossRef][Medline]
7. Alavi S., Namazie A., Calcaterra T. C., Wang M. B., Srivatsan E. S. Clinical application of fluorescence in situ hybridization for chromosome 11q13 analysis in head and neck cancer. Laryngoscope, 109: 874-879, 1999.[CrossRef][Medline]
8. Schuuring E. The involvement of the chromosome 11q13 region in human malignancies: cyclin D1 and EMS1 are two new candidate oncogenes—a review. Gene (Amst.), 159: 83-96, 1995.[CrossRef][Medline]
9. Meredith S. D., Levine P. A., Burns J. A., Gaffey M. J., Boyd J. C., Weiss L. M., Erickson N. L., Williams M. E. Chromosome 11q13 amplification in head and neck squamous cell carcinoma. Association with poor prognosis. Arch. Otolaryngol. Head Neck Surg., 121: 790-794, 1995.
10. Muller D., Millon R., Liderau R., Engelmann A., Bronner G., Eber M., Methlin G., Abecassis J. Frequent amplification of 11q13 DNA markers is associated with lymph node involvement in human head and neck squamous cell carcinomas. Eur. J. Cancer B Oral Oncol., 30B: 113-120, 1994.
11. Muller D., Millon R., Velten M., Bronner G., Jung G., Engelmann A., Flesch H., Eber M., Methlin G., Abecassis J. Amplification of 11q13 DNA markers in head and neck squamous cell carcinomas: correlation with clinical outcome. Eur. J. Cancer, 33A: 2203-2210, 1997.
12. Takes R. P., Baatenburg de Jong, R. J., Schuuring E., Hermans J., Vis A. A., Litvinov S., van Krieken J. H. J. M. Markers for assessment of nodal metastasis in laryngeal carcinoma. Arch. Otolaryngol. Head Neck Surg., 123: 412-419, 1997.
13. Jares P., Fernandez P. L., Campo E., Nadal A., Bosch F., Aiza G., Nayach I., Taraserra J., Cardesa A. PRAD-1/cyclin D1 gene amplification correlates with messenger RNA overexpression and tumor progression in human laryngeal carcinomas. Cancer Res., 54: 4813-4817, 1994.[Abstract/Free Full Text]
14. Michalides R., Van Veelen N., Hart A., Loftus B., Wientjens E., Balm A. Overexpression of cyclin D1 correlates with recurrence in a group of forty-seven operable squamous cell carcinomas of the head and neck. Cancer Res., 55: 975-978, 1995.[Abstract/Free Full Text]
15. Michalides R. J. A. M., van Veelen N. M. J., Kristel P. M. P., Hart A. A. M., Loftus B. M., Hilgers F. J. M., Balm A. J. M. Overexpression of cyclin D1 indicated a poor prognosis in squamous cell carcinoma of the head and neck. Arch. Otolaryngol. Head Neck Surg., 123: 479-502, 1997.
16. Bova R. J., Quinn D. I., Nankervis J. S., Cole I. E., Sheridan B. F., Jensen M. J., Morgan G. J., Hughes C. J., Sutherland R. L. Cyclin D1 and p16INK4A expression predict reduced survival in carcinoma of the anterior tongue. Clin. Cancer Res., 5: 2810-2819, 1999.[Abstract/Free Full Text]
17. Nogueira C. P., Dolan R. W., Gooey J., Byahatti S., Vaughn C. W., Fuleihan N. S., Grillona G., Baker E., Domanowski G. Inactivation of p53 and amplification of cyclin D1 correlate with clinical outcome in head and neck cancer. Laryngoscope, 108: 345-350, 1998.[CrossRef][Medline]
18. Patel A. S., Schechter G. L., Wasilenko W. J., Somers K. D. Overexpression of EMS1/cortactin in NIH3T3 fibroblasts causes increased cell motility and invasion in vitro. Oncogene, 16: 3227-3232, 1998.[CrossRef][Medline]
19. Rodrigo J. P., Ramos S., Lazo P. S., Alvarez I., Suárez C. Amplification of ERBB oncogenes in squamous cell carcinomas of the head and neck. Eur. J. Cancer, 32A: 2004-2010, 1996.
20. Frye R. A., Benz C. C., Liu E. Detection of amplified oncogenes by differential polymerase chain reaction. Oncogene, 4: 1153-1157, 1989.[Medline]
21. Schereiber G., Dubeau L. c-myc proto-oncogene amplification detected by polymerase chain reaction in archival human ovarian carcinomas. Am. J. Pathol., 137: 653-658, 1990.[Abstract]
22. Kaplan E. L., Meier P. Nonparametric estimation from incomplete observations. J. Am. Stat. Assoc., 53: 457-481, 1958.[CrossRef]
23. Peto R., Pike M. C., Armitage P. E., Breslow N. E., Cox D. R., Howard S. V., Mantel N., MacPherson K., Peto J., Smith P. G. Design and analysis of randomised clinical trials requiring prolonged observation of each patient. Br. J. Cancer, 34: 585-612, 1976.[Medline]
24. Cox D. R. Regression models and life tables. J. R. Stat. Soc. B, 34: 187-210, 1972.
25. Baldin V., Lukas J., Marcote M. J., Pagano M., Draetta G. Cyclin D1 is a nuclear protein required for cell-cycle progression in G₁. Genes Dev., 7: 812-821, 1993.[Abstract/Free Full Text]
26. Schuuring E., Verhoeven E., Litvinov S., Michalides R. J. The product of the EMS1 gene, amplified and overexpressed in human carcinomas, is homologous to a v-src substrate and is located in cell-substratum contact sites. Mol. Cell. Biol., 13: 2891-2898, 1993.[Abstract/Free Full Text]
27. Karlseder J., Zeillinger R., Schneeberger C., Czerwenka K., Speiser P., Kubista E., Birnbaum D., Gaudray P., Theillet C. Patterns of DNA amplification at band q13 of chromosome 11 in human breast cancer. Genes Chromosomes Cancer, 9: 42-48, 1994.[Medline]
28. Gleich L. L., Li Y. Q., Wang X., Stambrook P., Gluckman J. L. Variable genetic alterations and survival in head and neck cancer. Arch. Otolaryngol. Head Neck Surg., 125: 949-952, 1999.

This article has been cited by other articles:

				B. Singh and D. G. Pfister Individualized Treatment Selection in Patients With Head and Neck Cancer: Do Molecular Markers Meet the Challenge? J. Clin. Oncol., July 1, 2008; 26(19): 3114 - 3116. [Full Text] [PDF]

				X.-Y. Lu, Y. Lu, Y.-J. Zhao, K. Jaeweon, J. Kang, L. Xiao-Nan, G. Ge, R. Meyer, L. Perlaky, J. Hicks, et al. Cell Cycle Regulator Gene CDC5L, a Potential Target for 6p12-p21 Amplicon in Osteosarcoma Mol. Cancer Res., June 1, 2008; 6(6): 937 - 946. [Abstract] [Full Text] [PDF]

				J. H. Gibcus, L. Menkema, M. F. Mastik, M. A. Hermsen, G. H. de Bock, M.-L. F. van Velthuysen, R. P. Takes, K. Kok, C. A. Alvarez Marcos, B. F.A.M. van der Laan, et al. Amplicon Mapping and Expression Profiling Identify the Fas-Associated Death Domain Gene as a New Driver in the 11q13.3 Amplicon in Laryngeal/Pharyngeal Cancer Clin. Cancer Res., November 1, 2007; 13(21): 6257 - 6266. [Abstract] [Full Text] [PDF]

				P. Timpson, A. S. Wilson, G. M. Lehrbach, R. L. Sutherland, E. A. Musgrove, and R. J. Daly Aberrant Expression of Cortactin in Head and Neck Squamous Cell Carcinoma Cells Is Associated with Enhanced Cell Proliferation and Resistance to the Epidermal Growth Factor Receptor Inhibitor Gefitinib Cancer Res., October 1, 2007; 67(19): 9304 - 9314. [Abstract] [Full Text] [PDF]

				S. Skvortsov, I. Skvortsova, T. Stasyk, N. Schiefermeier, A. Neher, A. R. Gunkel, G. K. Bonn, L. A. Huber, P. Lukas, C. M. Pleiman, et al. Antitumor activity of CTFB, a novel anticancer agent, is associated with the down-regulation of nuclear factor-{kappa}B expression and proteasome activation in head and neck squamous carcinoma cell lines Mol. Cancer Ther., June 1, 2007; 6(6): 1898 - 1908. [Abstract] [Full Text] [PDF]

				E. S. Clark, A. S. Whigham, W. G. Yarbrough, and A. M. Weaver Cortactin Is an Essential Regulator of Matrix Metalloproteinase Secretion and Extracellular Matrix Degradation in Invadopodia Cancer Res., May 1, 2007; 67(9): 4227 - 4235. [Abstract] [Full Text] [PDF]

				M.-L. Luo, X.-M. Shen, Y. Zhang, F. Wei, X. Xu, Y. Cai, X. Zhang, Y.-T. Sun, Q.-M. Zhan, M. Wu, et al. Amplification and Overexpression of CTTN (EMS1) Contribute to the Metastasis of Esophageal Squamous Cell Carcinoma by Promoting Cell Migration and Anoikis Resistance Cancer Res., December 15, 2006; 66(24): 11690 - 11699. [Abstract] [Full Text] [PDF]

				B. L. Rothschild, A. H. Shim, A. G. Ammer, L. C. Kelley, K. B. Irby, J. A. Head, L. Chen, M. Varella-Garcia, P. G. Sacks, B. Frederick, et al. Cortactin Overexpression Regulates Actin-Related Protein 2/3 Complex Activity, Motility, and Invasion in Carcinomas with Chromosome 11q13 Amplification Cancer Res., August 15, 2006; 66(16): 8017 - 8025. [Abstract] [Full Text] [PDF]

				Y. Kasugai, H. Tagawa, Y. Kameoka, Y. Morishima, S. Nakamura, and M. Seto Identification of CCND3 and BYSL as Candidate Targets for the 6p21 Amplification in Diffuse Large B-Cell Lymphoma Clin. Cancer Res., December 1, 2005; 11(23): 8265 - 8272. [Abstract] [Full Text] [PDF]

				P. Timpson, D. K. Lynch, D. Schramek, F. Walker, and R. J. Daly Cortactin Overexpression Inhibits Ligand-Induced Down-regulation of the Epidermal Growth Factor Receptor Cancer Res., April 15, 2005; 65(8): 3273 - 3280. [Abstract] [Full Text] [PDF]

				Y. Chen, G. Shi, W. Xia, C. Kong, S. Zhao, A. F. Gaw, E. Y. Chen, G. P. Yang, A. J. Giaccia, Q.-T. Le, et al. Identification of Hypoxia-Regulated Proteins in Head and Neck Cancer by Proteomic and Tissue Array Profiling Cancer Res., October 15, 2004; 64(20): 7302 - 7310. [Abstract] [Full Text] [PDF]

				P. Matar, F. Rojo, R. Cassia, G. Moreno-Bueno, S. Di Cosimo, J. Tabernero, M. Guzman, S. Rodriguez, J. Arribas, J. Palacios, et al. Combined Epidermal Growth Factor Receptor Targeting with the Tyrosine Kinase Inhibitor Gefitinib (ZD1839) and the Monoclonal Antibody Cetuximab (IMC-C225): Superiority Over Single-Agent Receptor Targeting Clin. Cancer Res., October 1, 2004; 10(19): 6487 - 6501. [Abstract] [Full Text] [PDF]

				Q.-T. Le and A. J. Giaccia Therapeutic Exploitation of the Physiological and Molecular Genetic Alterations in Head and Neck Cancer Clin. Cancer Res., October 1, 2003; 9(12): 4287 - 4295. [Abstract] [Full Text] [PDF]

				D. K. Lynch, S. C. Winata, R. J. Lyons, W. E. Hughes, G. M. Lehrbach, V. Wasinger, G. Corthals, S. Cordwell, and R. J. Daly A Cortactin-CD2-associated Protein (CD2AP) Complex Provides a Novel Link between Epidermal Growth Factor Receptor Endocytosis and the Actin Cytoskeleton J. Biol. Chem., June 6, 2003; 278(24): 21805 - 21813. [Abstract] [Full Text] [PDF]

				K. Freier, S. Joos, C. Flechtenmacher, F. Devens, A. Benner, F. X. Bosch, P. Lichter, and C. Hofele Tissue Microarray Analysis Reveals Site-specific Prevalence of Oncogene Amplifications in Head and Neck Squamous Cell Carcinoma Cancer Res., March 15, 2003; 63(6): 1179 - 1182. [Abstract] [Full Text] [PDF]

				B.-Z. Yuan, X. Zhou, D. B. Zimonjic, M. E. Durkin, and N. C. Popescu Amplification and Overexpression of the EMS 1 Oncogene, a Possible Prognostic Marker, in Human Hepatocellular Carcinoma J. Mol. Diagn., February 1, 2003; 5(1): 48 - 53. [Abstract] [Full Text] [PDF]

				T. Ueno, A. Tangoku, S. Yoshino, T. Abe, H. Toshimitsu, T. Furuya, S. Kawauchi, A. Oga, M. Oka, and K. Sasaki Gain of 5p15 Detected by Comparative Genomic Hybridization as an Independent Marker of Poor Prognosis in Patients with Esophageal Squamous Cell Carcinoma Clin. Cancer Res., February 1, 2002; 8(2): 526 - 533. [Abstract] [Full Text] [PDF]

				Y. Li, M. Tondravi, J. Liu, E. Smith, C. C. Haudenschild, M. Kaczmarek, and X. Zhan Cortactin Potentiates Bone Metastasis of Breast Cancer Cells Cancer Res., September 1, 2001; 61(18): 6906 - 6911. [Abstract] [Full Text] [PDF]

This Article Abstract Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Rodrigo, J. P. Articles by Suárez, C. Search for Related Content PubMed PubMed Citation Articles by Rodrigo, J. P. Articles by Suárez, C.