COX-2 expression in dysplasia of the head and neck: correlation with elF4E.

BACKGROUND: COX-2 inhibitors have shown promise in chemoprevention of epithelial tumors. eIF4E is a biomarker that has identified individuals at high risk for relapse after definitive treatment for head and neck squamous cell cancer (HNSCC). Hence, the authors wanted to determine if COX-2 is expressed in dysplasia of the head and neck and to study the correlation of expression of COX-2 with eIF4E as a potential surrogate endpoint for determining response to COX-2 inhibitors. METHODS: The authors studied the expression of COX-2 and eIF4E in normal epithelium (n = 8), dysplasia (n = 51), mucosa adjacent to tumors (n = 11), and cancer of the head and neck (n = 19) using immunohistochemistry. In addition, Western blot analysis was performed on a subset of the above patient samples and HNSCC cell lines. RESULTS: Immunohistochemical analysis showed expression of COX-2 and eIF4E in all cancers and no expression in normal tissues. In dysplastic epithelium, there was a significant correlation between the expression of eIF4E and COX-2 for all groups of dysplasia combined (chi-square = 40.3, P < 0.001). A Cochran-Armitage trend test showed a significant increase in the proportion of cases that expressed both molecular markers with increasing grades of dysplasia (P = 0.001). Western blot analysis showed increased expression of COX-2 and eIF4E in tumors compared with adjacent mucosa. All three HNSCC cell lines analyzed had increased expression of eIF4E, although only two had increased COX-2 expression. CONCLUSIONS: Expression of COX-2 in dysplasia suggested that COX-2 inhibitors may play a role in chemoprevention of head and neck cancers and that the correlation of Cox-2 with eIF4E indicates that eIF4E can be a potential surrogate marker in chemoprevention trials.

A diagnostic and therapeutic approach to paragangliomas of the larynx.

BACKGROUND: Differentiating paragangliomas from moderately differentiated neuroendocrine carcinoma in the larynx is a difficult management problem. As the biological behavior of these 2 entities is different, we developed an algorithm for the preoperative diagnosis and treatment of this disease. DESIGN: The sample case from which the algorithm was developed consisted of a 69-year-old man who was transferred to us after tracheostomy and an attempt at biopsy for airway obstruction secondary to a vascular mass. Biopsy resulted in substantial bleeding. Flexible laryngoscopy showed a vascular mass of the supraglottis. A computed tomographic scan showed 2 vascular masses at the carotid bifurcation and in the larynx. An arteriogram confirmed synchronous vascular tumors. RESULTS: The arteriogram showed the superior thyroid artery to be the major feeder vessel to this mass, a situation commonly seen in paragangliomas but not other neuroendocrine tumors. The presence of synchronous lesions and a vascular mass based on the superior thyroid artery helped differentiate paraganglioma from the other neuroendocrine tumors. As the biological behavior of paragangliomas is relatively benign, we performed a conservative supraglottic laryngectomy and excision of the carotid body tumor. Histologic diagnosis and immunohistochemical analysis confirmed the diagnosis of paraganglioma. CONCLUSIONS: The vascular nature of neuroendocrine tumors prevents preoperative pathological diagnosis. Radiologic features demonstrating a vascular mass with a dominant feeder vessel by the superior or inferior thyroid artery may help in the clinical diagnosis of paragangliomas of the larynx. Since paragangliomas are rarely malignant, a conservative surgical procedure should suffice.

Clinical application of botulinum toxin in otolaryngology, head and neck practice (brief review).

Botulinum toxin (Botox) is useful in controlling the symptoms of patients with movement disorders. Application of Botox serves to (1) inhibit hypertonicity, (2) enhance the action of the antagonistic muscles, and (3) avoid an impingement in order to reestablish "the balance of forces". In accordance with the principles mentioned above, Botox can be used to treat dystonias of the larynx (adductor laryngeal spasmodic dysphonia, abductor laryngeal spasmodic dysphonia), laryngeal granulomas, laryngeal joint dislocation, cricopharyngeal spasm, and posterior glottic synechiae. In addition, extra-laryngeal disorders such as blepharospasm, hemifacial spasm, oromandibular dystonia, and spasmodic torticollis respond well to Botox. The effects of Botox are reversible and have specific localized activity. Hence, Botox has served as a powerful diagnostic method in exploring the underlying mechanism of various types of dystonias and provides some therapeutic benefits before pursuing surgical options. Here we review the literature and describe our experiences with Botox, including such topics as preparing and storing Botox, identifying the target muscles under EMG-guidance, choosing an appropriate dose, and outlining the applications of Botox in Otolaryngology, Head and Neck Surgery practice.

Pott's puffy tumor and epidural abscess arising from dental sepsis: a case report.

OBJECTIVE: To present an unusual case of two uncommon cranial complications of frontal sinusitis: Pott's puffy tumor and epidural abscess arising from frontal sinusitis of dental origin, and also two systemic complications of sinusitis: septicemia and empyema, all occurring in an immunocompetent patient. STUDY DESIGN: A 21-year-old man presented with a scalp swelling and epidural abscess. Magnetic resonance imaging and computed tomographic scans revealed unilateral opacification of the frontal sinus and an epidural abscess with a direct connection to the scalp abscess. Further history revealed that his symptoms occurred coincidentally with a tooth extraction 2 months before, and he was hospitalized soon after the tooth extraction for sepsis and a lung abscess. METHODS: A combined neurosurgical and otolaryngologic approach was required to treat the sinusitis and the associated epidural and scalp abscess. RESULTS: Cultures returned as Streptococcus intermedius from all three sites. The patient was free of disease at the 3-month follow-up. CONCLUSIONS: Odontogenic maxillary sinusitis is well documented; however, there is little reported of frontal sinusitis arising from dental disease. The prevalence of sinusitis of dental origin will be reviewed, including the microbiology of this particularly virulent organism that persisted despite earlier treatment with ampicillin. Also, the current thoughts on management of these cases will be discussed with particular reference to local therapy for sinusitis in addition to systemic treatment with antibiotics.

Correlation of p53 and the proto-oncogene eIF4E in larynx cancers: prognostic implications.

p53 abnormalities constitute the most frequent genetic alterations identified in larynx cancers. p53 overexpression in histologically "tumor-free" surgical margins correlates with a high recurrence rate. However, only 50-60% of tumors overexpress p53. The tumor marker eIF4E is overexpressed in 100% of larynx cancers, and overexpression of eIF4E in histologically "tumor-free" margins predicts a significantly higher recurrence. We undertook this study to correlate the expression of p53 and eIF4E in the tumors and surgical margins of squamous cell cancers of the larynx and to determine their prognostic value. A retrospective analysis was performed on 54 patients who underwent surgery for squamous cell cancers of the larynx. Patient and tumor characteristics were reviewed, and the time to recurrence was noted. Paraffin-embedded sections from the tumors and surgical margins were immunostained with antibodies to eIF4E and p53, and a qualitative analysis was performed. All 54 patients (100%) overexpressed eIF4E in the primary tumor, whereas 25 of 53 patients (47%) were p53 positive. Thirty-two of the 54 patients (59%) had eIF4E-positive margins. All 6 of 53 patients (11%) with p53-positive margins also overexpressed eIF4E in the margins. There was a significant correlation between p53 and eIF4E being positive in the margins (Spearman's correlation coefficient, P = 0.03). Twenty-one of the 25 patients (84%) that recurred, including the 6 patients with p53-positive margins, had eIF4E-positive margins. Hence, although the univariate analysis showed that nodal status and both eIF4E and p53 expression in the margins were significant predictors of recurrence (P < 0.05), in the multivariate analyses only nodal status (P < 0.001) and eIF4E in the margins (P < 0.001) were significant predictors of recurrence. Kaplan-Meier analysis demonstrated that the disease-free intervals for eIF4E-positive margins were significantly shorter than eIF4E-negative margins (P = 0.0007). There was no additional effect to the combination of positive p53 and eIF4E margins (P = 0.21). The overexpression of eIF4E in the margins appears to be a more sensitive indicator of recurrence and may be an earlier event in the process of tumorigenesis than p53.

Antisense RNA to eIF4E suppresses oncogenic properties of a head and neck squamous cell carcinoma cell line.

OBJECTIVE: The translation initiation factor eIF4E is elevated in all head and neck squamous cell cancers (HNSCCs) and appears to be essential in the progression of solid tumors. Overexpression of eIF4E results in preferential upregulation of two angiogenic factors, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (FGF-2). We wanted to determine whether reducing eIF4E in a HNSCC cell line could suppress its oncogenic properties and in turn decrease expression of VEGF and FGF-2. METHODS: Levels of eIF4E protein expression were determined in a panel of HNSCC cell lines. An episomal vector containing antisense RNA to eIF4E was used to reduce the eIF4E level in one of these cell lines, FaDu. After a stable transfection, Western blot analysis was performed to determine the level of eIF4E and FGF-2 reduction, while an enzyme-linked immunosorbent assay (ELISA) was used to determine the level of VEGF reduction. In vitro and in vivo experiments were performed to determine whether there was a reversion in the tumorigenic properties of the FaDu cells. RESULTS: All six cell lines had elevated levels of eIF4E compared with Detroit 551, a normal cell line. Reducing eIF4E expression via antisense RNA suppressed both the tumorigenic and angiogenic properties of the FaDu cells, as demonstrated by loss of capacity to grow in soft agar, reduced expression of angiogenic factors, and loss of tumorigenicity in nude mice. CONCLUSIONS: Antisense RNA therapy to eIF4E can potentially be used as adjuvant therapy for head and neck cancers, particularly in cases in which elevated eIF4E is found in the surgical margins.

Reduced F-18 fluorodeoxyglucose uptake within marrow after external beam radiation.

PURPOSE: The authors examined the delayed effects of external beam radiation on the metabolism of intravertebral marrow cavities. MATERIALS AND METHODS: Two neurologically normal persons who had completed a full course of external beam radiotherapy for squamous cell carcinoma of the head and neck had positron emission tomography (PET) scans with F-18 fluorodeoxyglucose (FDG) to identify residual neoplasm. RESULTS: Abnormally low uptake of F-18 FDG was noted within the central marrow cavities of the cervical vertebral bodies in both cases. CONCLUSIONS: Diminished metabolic activity within irradiated marrow can be revealed by F-18 FDG PET imaging techniques. Further correlative MRI PET studies are needed to determine if this postradiation phenomenon is related to fatty infiltration of the marrow cavity.

Progressive amplification and overexpression of the eukaryotic initiation factor 4E gene in different zones of head and neck cancers.

PURPOSE: Eukaryotic initiation factor 4E (eIF4E) binds to mRNA as the initial rate-limiting step in protein synthesis. Amplification and overexpression of the eIF4E gene has been associated with malignant transformation. The objectives of this study were to 1) quantify the eIF4E gene in head and neck squamous cell carcinoma (HNSCC) specimens, 2) quantify eIF4E protein elevation and examine its association with eIF4E gene amplification, and 3) determine whether there is progression in eIF4E gene amplification and protein overexpression in the tumor-free resection margin, the transition zone, and the tumor core of HNSCC specimens. MATERIALS AND METHODS: Eighteen HNSCC specimens were divided into three zones: 1) tumor core; 2) transition zone; and 3) "tumor-free" margin. Competitive polymerase chain reaction was performed to determine eIF4E gene copy number. eIF4E protein expression was quantified using Western blot analysis. RESULTS: All 18 HNSCC specimens tested had significant eIF4E gene amplification (4.3+/-1.2; P < .05). In contrast, none of the 10 benign specimens from noncancer patients had any eIF4E gene amplification (1.1+/-0.5). In the 12 HNSCC specimens examined for the three zones, the tumor core and transition zone showed eIF4E gene amplification (5.2+/-1.1 and 3.5+/-0.9, respectively) compared with the "tumor-free" margin (2.1+/-1.1; P < .05). The tumor core and transition zone showed significant efF4E protein elevation (15.5+/-9.3, 4.4+/-4.6, respectively) compared with the "tumor-free" margin (0.9+/-0.5, P < .05). CONCLUSIONS: The eIF4E gene is amplified and overexpressed in HNSCC. Amplification and elevation of eIF4E were highest in the tumor core, intermediate in the transition zone, and lowest in the tumor-free margin. There appears to be progression of eIF4E gene amplification and overexpression from the "tumor-free" margin to the tumor core.

Detection of the proto-oncogene eIF4E in larynx and hypopharynx cancers.

BACKGROUND: The proto-oncogene eIF4E has been found to be elevated in head and neck squamous cell carcinomas. In an earlier prospective study overexpression of eIF4E, detected by Western blot analysis, in histologically normal surgical margins correlated with an increased local-regional recurrence rate during a 1-year follow-up. OBJECTIVE: To test the reverse hypothesis that absence of overexpression of eIF4E in the surgical margins is a predictor for long-term survival in patients with squamous cell carcinoma of the head and neck. DESIGN: A retrospective analysis was performed on 31 patients who underwent surgery for squamous cell carcinoma of the larynx or hypopharynx. Immunohistochemical analysis was used to detect eIF4E on paraffin embedded sections of the tumor and the histologically negative surgical margins. RESULTS: All 31 patients overexpressed eIF4E in the tumors. Thirteen patients had no detectable level of eIF4E in the margins, and only 1 had a local-regional recurrence. The average disease-free interval in this group of patients was 82.08 months. The remaining 18 patients all overexpressed eIF4E in the surgical margins (eIF4E score range, 5-80). Twelve (67%) of these patients developed a recurrence; the average disease-free interval was 31.95 months. Cox regression analysis showed that eIF4E in the margin (P= .01), nodes (P= .06), site (P= .02), and age (P= .02) had significant effects on the disease-free interval. The Kaplan-Meier survival curves were significantly different for eIF4E-positive and eIF4E-negative margins (P = .002). CONCLUSIONS: eIF4E in the surgical margins is an independent prognostic factor and its absence in surgical margins may predict long-term survival. Detecting eIF4E in the margins may improve survival by determining which patients would benefit from further resection or adjuvant therapy.

Competitive PCR to detect eIF4E gene amplification in head and neck cancer.

BACKGROUND: The protein eukaryotic initiation factor 4E (elF4E) binds to messenger ribonucleic acid (mRNA) as the initial step in protein synthesis. Overexpression of elF4E results in upregulation of specific proteins essential to cell growth and division. Overexpression of elF4E has been found in head and neck squamous cell carcinoma (HNSCC) and breast carcinoma. This study's purpose is to determine whether elF4E overexpression is present and associated with elF4E gene amplification in HNSCC. METHODS: Competitive polymerase chain reaction (PCR) was performed on eight HNSCC and seven intraoral benign lesions to determine the copy number of elF4E relative to a reference gene, gastrin. Western blots were performed to quantify elF4E protein expression. RESULTS: All eight HNSCC specimens demonstrated significant (p < .005) overexpression of elF4E protein (14.1+/-10.4) and elF4E gene amplification (4.5+/-1.2). Benign tissue did not exhibit elF4E protein overexpression or gene amplification. CONCLUSIONS: Overexpression and associated gene amplification of elF4E were present in HNSCC but not in benign tissue. Gene amplification of elF4E may be an important mechanism for elF4E overexpression.

Absence of mRNA encoding estrogen receptor in the rat cochlea.

Based on changes in hearing thresholds and tinnitus that are co-related with the menstrual cycle, it has been suggested that the cochlea may respond directly to estrogen. For this to occur, the cochlea should express estrogen receptors. In situ mRNA hybridization was performed on normal female rat cochleas, using radiolabeled RNA probes complementary to mRNA encoding estrogen receptor, to determine whether estrogen receptors are present in the cochlea. Strong hybridization of the riboprobes to sections of uterus and hypothalamus indicated that the technique detected estrogen receptor mRNA. No hybridization to any cochlear tissues was observed. The results indicate that estrogen receptors are not expressed on cochlear cells, at least in rats. This in turn suggests that variation in cochlear responses during the estrus cycle are not the result of the direct effect of estrogen on the cochlea. Such variation may, however, be caused by systemic changes in fluid regulation induced by estrogen receptors at a distant site, or by other hormone receptors.

Effects of postoperative external beam irradiation after carotid artery stripping of nodal metastasis.

PURPOSE: The objective of this study was to determine the effects of postoperative external beam irradiation (PEBI) on patients after stripping the carotid vessel of nodal metastasis. PATIENTS AND METHODS: Of 245 patients who underwent radical neck dissection for cancer of the upper aerodigestive tract between 1981 and 1995, 13 patients with nodal metastasis adherent to the carotid artery (NMCA) received a full course of PEBI after the tumor was peeled off the carotid vessel. Patterns of treatment failure, survival, and serious morbidity were evaluated. RESULTS: At the time of last follow-up (range, 3 to 125 months), fewer than half of the patients (46%) were free of disease, and two patients (15%) had developed distant metastases. The estimated 2-year survival rate was 23%. There were no intraoperative, postoperative, or post-radiation therapy complications. CONCLUSION: PEBI after carotid artery stripping of tumor can achieve locoregional disease control in select patients without an increased risk of vessel rupture. However, if the prognosis for patients with NMCA is to improve, other effective management strategies need to be investigated.

Elevated expression of eIF4E and FGF-2 isoforms during vascularization of breast carcinomas.

The translation initiation factor eIF4E is a novel protooncogene found over expressed in most breast carcinomas (Kerekatte et al., 1995), but the pathology where this elevation is initially manifested and its possible role in cancer progression are unknown. We report that eIF4E is markedly increased in vascularized malignant ductules of invasive carcinomas, whereas necrotic and avascular ductal carcinomas in situ display significantly lower levels. eIF4E facilitates the synthesis of FGF-2, a powerful tumor angiogenic factor. Conversely, reducing eIF4E with antisense RNA in MDA-435 cells suppresses their tumorigenic and angiogenic properties, consistent with loss of FGF-2 synthesis. These findings suggest a causal role for eIF4E in tumor vascularization.

Detection of the proto-oncogene eIF4E in surgical margins may predict recurrence in head and neck cancer.

Head and neck squamous cell cancers (HNSCC) have a high local recurrence rate due to incomplete tumor resection. The use of molecular markers to establish surgical margins may decrease local recurrence. Surgical margins are determined by histopathologic analysis on frozen sections. We postulate that genetic and molecular changes precede gross histologic alterations. Tumor markers may improve the reliability of pathology examination, but those evaluated to date lack the sensitivity needed for routine clinical use. Western blot analysis showed elevated eIF4E in all 26 HNSCC in contrast to its low expression in benign lesions. Surgical margins were analysed for eIF4E in 23 patients. Twelve patients showed elevated eIF4E in histologically negative margins. Cancer has recurred in 5 of the 12 patients as opposed to none of the 11 patients with eIF4E negative margins (P= 0.02, Log rank test). This is the first report of eIF4E in HNSCC, as a sensitive and specific marker for HNSCC, with potential for defining clear resection margins. The correlation between elevated levels of eIF4E at the margins and recurrence highlights its ability to detect malignant cells prior to clear-cut alterations in morphology. The accuracy and simplicity of these assays underscore the usefulness of eIF4E in managing HNSCC.