Fernando Walder

Overexpression of Fos-related antigen-1 in head and neck squamous cell carcinoma

The activating protein‐1 (AP‐1) family of transcription factors has been implicated in the control of proliferation and differentiation of keratinocytes, but its role in malignant transformation is not clear. The aim of this study is to assess the pattern of mRNA expression of jun‐fos AP‐1 family members in 45 samples of head and neck squamous cell carcinomas (HNSCC) and matched adjacent mucosa by means of Northern blot analysis. Transcripts of all family members were identified, except for JunB that was detected only by means of reverse transcription polymerase chain reaction. Neither c‐Fos nor JunD or FosB mRNA differed between tumours and normal tissues. We observed a strong Fos‐related antigen‐1 (Fra‐1) and Fra‐2 expression, but only Fra‐1 mRNA densitometric values were higher in tumour, compared to normal adjacent mucosa (t‐test, P = 0.006). A direct relationship between the positive expression of Fra‐1 mRNA, above tumour median, was associated with the presence of compromised lymph nodes (Fischer exact test, P = 0.006). In addition, Fra‐1 protein staining was assessed in a collection of 180 tumours and 29 histologically normal samples adjacent to tumours in a tissue array. Weak reactivity, restricted to the basal cell layer, was detected in 79% of tumour adjacent normal tissues, opposed to the intense reactivity of cancer tissues. In the subgroup of oral cancers, we have observed a shift in Fra‐1 immunoreactivity, as long as the number of patients in each category, cytoplasmic or nuclear/cytoplasmic staining, was analysed (Fischer exact test, P = 0.0005). Thus, Fra‐1 gene induction and accumulation of Fra‐1 protein may contribute to the neoplastic phenotype in HNSCC.

Perioperative frozen section examination in parotid gland tumors

CONTEXT The minimal recommended surgical approach to parotid tumors is partial parotidectomy with resection of the superficial lobe of the gland. Histologic diagnosis prior to surgery is not possible, as incisional biopsies are contraindicated due to the possibility of facial nerve injury or incomplete tumor resection. Thus, the biopsies tend to be perioperative. OBJECTIVE To compare the results of frozen section examination with the definitive pathological diagnosis. DESIGN Accuracy study by retrospective analysis. SETTING Head and Neck Surgery Service of Heliópolis Hospital, São Paulo, Brazil. SAMPLE 153 cases of parotid gland tumors treated between 1977 and 1994. DIAGNOSTIC TEST Frozen section and pathological diagnosis. MAIN MEASUREMENTS Sensibility and specificity of the frozen section examination. RESULTS Frozen section study diagnosed 19 (12.4%) malignant and 127 (83.7%) benign tumors. Sensitivity of the frozen sections for malignancy was 61.5% (95% CI 54 to 69%) and specificity was 98% (95% CI 94 to 100%), and this result is comparable to the literature. CONCLUSIONS We consider that frozen section examination for salivary gland tumors is not sufficient on its own for deciding on the best management. Their interpretation must be correlated with clinical and intraoperative findings, in association with the surgeons experience.

Smad2 and Smad6 as predictors of overall survival in oral squamous cell carcinoma patients

BackgroundTo test if the expression of Smad1-8 mRNAs were predictive of survival in patients with oral squamous cell carcinoma (SCC).Patients and MethodsWe analyzed, prospectively, the expression of Smad1-8, by means of Ribonuclease Protection Assay in 48 primary, operable, oral SCC. In addition, 21 larynx, 10 oropharynx and 4 hypopharynx SCC and 65 matched adjacent mucosa, available for study, were also included. For survival analysis, patients were categorized as positive or negative for each Smad, according to median mRNA expression. We also performed real-time quantitative PCR (QRTPCR) to asses the pattern of TGFβ1, TGFβ2, TGFβ3 in oral SCC.ResultsOur results showed that Smad2 and Smad6 mRNA expression were both associated with survival in Oral SCC patients. Cox Multivariate analysis revealed that Smad6 positivity and Smad2 negativity were both predictive of good prognosis for oral SCC patients, independent of lymph nodal status (P = 0.003 and P = 0.029, respectively). In addition, simultaneously Smad2- and Smad6+ oral SCC group of patients did not reach median overall survival (mOS) whereas the mOS of Smad2+/Smad6- subgroup was 11.6 months (P = 0.004, univariate analysis). Regarding to TGFβ isoforms, we found that Smad2 mRNA and TGFβ1 mRNA were inversely correlated (p = 0.05, R = -0.33), and that seven of the eight TGFβ1+ patients were Smad2-. In larynx SCC, Smad7- patients did not reach mOS whereas mOS of Smad7+ patients were only 7.0 months (P = 0.04). No other correlations were found among Smad expression, clinico-pathological characteristics and survival in oral, larynx, hypopharynx, oropharynx or the entire head and neck SCC population.ConclusionSmad6 together with Smad2 may be prognostic factors, independent of nodal status in oral SCC after curative resection. The underlying mechanism which involves aberrant TGFβ signaling should be better clarified in the future.

Head and neck squamous cell carcinoma in childhood

BACKGROUND Squamous cell carcinoma (SCC) of the head and neck region is rare in young patients and even less frequent in children 15 years or younger children. The patients reported in the literature are isolated cases and their management is always difficult because there is no large experience or a convincing theory to support treatment decisions for every child. PROCEDURES AND RESULTS Four patients aged 15 years or younger were treated for SCC of head and neck between 1977 and 1995 at the Head and Neck Service of Heliópolis Hospital, São Paulo, Brazil, and with this paper we are reporting our experience with their treatment, including a genetic investigation in two cases (immunohistochemical analysis using monoclonal antibodies against p53 and c-erbB-2 oncogenes). These patients had no history of tobacco or ethanol abuse and no history of cancer in their families. Many authors attribute an unusual aggressiveness to SCC in childhood, with propensity to locoregional recurrence and high death rates and therefore, propose aggressive multidisciplinary therapy. Our cases, with the exception of one, had an early diagnosis and were treated using the same method we use for adults; the results were very good. CONCLUSIONS We recommend for these cases the same protocol as for older patients. In these cases, however, the primary lesion is resected with a safety margin which is usually 2 to 3 mm larger than usual safety margins and selective neck dissection is routinely indicated. This management is adopted in an attempt to avoid postoperative irradiation which may prove to be dangerous in the future for young patients. On the other hand, we recognize that due to a small number de patients, definitive treatment recommendations cannot be made at this time.

Four-gene expression model predictive of lymph node metastases in oral squamous cell carcinoma

Abstract Background. Previous knowledge of cervical lymph node compromise may be crucial to choose the best treatment strategy in oral squamous cell carcinoma (OSCC). Here we propose a set four genes, whose mRNA expression in the primary tumor predicts nodal status in OSCC, excluding tongue. Material and methods. We identified differentially expressed genes in OSCC with and without compromised lymph nodes using Differential Display RT-PCR. Known genes were chosen to be validated by means of Northern blotting or real time RT-PCR (qRT-PCR). Thereafter we constructed a Nodal Index (NI) using discriminant analysis in a learning set of 35 patients, which was further validated in a second independent group of 20 patients. Results. Of the 63 differentially expressed known genes identified comparing three lymph node positive (pN +) and three negative (pN0) primary tumors, 23 were analyzed by Northern analysis or RT-PCR in 49 primary tumors. Six genes confirmed as differentially expressed were used to construct a NI, as the best set predictive of lymph nodal status, with the final result including four genes. The NI was able to correctly classify 32 of 35 patients comprising the learning group (88.6%; p = 0.009). Casein kinase 1alpha1 and scavenger receptor class B, member 2 were found to be up regulated in pN + group in contrast to small proline-rich protein 2B and Ras-GTPase activating protein SH3 domain-binding protein 2 which were upregulated in the pN0 group. We validated further our NI in an independent set of 20 primary tumors, 11 of them pN0 and nine pN + with an accuracy of 80.0% (p = 0.012). Conclusions. The NI was an independent predictor of compromised lymph nodes, taking into the consideration tumor size and histological grade. The genes identified here that integrate our “Nodal Index” model are predictive of lymph node metastasis in OSCC.

Glycosphingolipid expression in squamous cell carcinoma of the upper aerodigestive tract

UNLABELLED Glycosphingolipids are integral constituents of cellular membrane, arranged in rafts, and with neoplastic cell anti-social behavior, like uncontrolled cell growth, invasiveness, and metastatic potential. AIM However, there are few studies about glycosphingolipids (GSL) expression in squamous cell carcinoma (SCC). Since GSL are known to be tumor-associated markers we decided to perform a prospective study on the GSL profiles of SCC. METHOD Specimens of 33 SCC and normal mucosa were obtained and GSLs were extracted and purified by reverse-phase chromatography on C18 column and alkaline hydrolysis in methanol. GSLs were quantified using densitometry of orcinol-stained HPTLC plates. RESULT A significant increase of GSLs in SCC (3.57 microg/mg) was observed as compared to normal mucosa (1.92 microg/mg). In SCC, an increase of 2 to 3 times in the amounts of CDH, CTH, Globoside, and GM3 was observed in comparison to normal mucosa. The identification of GM3 as well as its increased expression in SCC was confirmed unequivocally by HPTLC immunostaining and indirect immunofluorescence using MAb DH2 (anti-GM3). BY analyzing SCC and normal mucosa CMHs by GC/MS, normal mucosa expresses only glucosylceramide whereas SCC cells express both glucosylceramide and galactosylceramide. CONCLUSION The increase in the amount of GSLs in tumor tissue may represent changes of cell membrane microdomains resulting from the malignant transformation process, which is responsible for greater cell-cell or cell-matrix interaction thereby increasing their potential for infiltration and metastasis.

The role of transoral robotic surgery in the management of oropharyngeal cancer.

Purpose of review The rising incidence of oropharyngeal squamous cell carcinoma (OPSCC), in large part as a result of the human papillomavirus (HPV), has driven a movement for the change in the management strategies. Renewed interest in minimally invasive approaches of endoscopic head and neck surgery led to introduction of transoral surgery, including transoral robotic surgery (TORS). Recent findings Several recent studies, based on large multi-institutional studies and systematic reviews of the literature, have shown excellent oncologic and functional outcomes with TORS for OPSCC. Also, a growing amount of clinical evidence supports the use of TORS in the management of carcinoma of unknown primary site and in selected patients with recurrent OPSCC with acceptable oncologic and better functional outcomes in comparison with traditional surgical approaches. Comparative studies with other therapeutic modalities (conventional surgical and nonsurgical) showed that TORS can be used to treat OPSCC, reducing morbidity and treatment costs, while providing equivalent oncologic results. Summary Large and robust data available in the literature supports the role of TORS within the multidisciplinary treatment paradigm for the management of OPSCC. Information from ongoing randomized clinical trials comparing TORS with and without dose-reduced radiotherapy or with and without intensified adjuvant treatment for high-risk OPSCC patients is necessary to determine the role of de-escalation of therapy in the era of HPV and OPSCC.

Transoral robotic supraglottic partial laryngectomy: report of the first Brazilian case.

In the past decade, we have witnessed the introduction and dissemination of transoral robotic surgery for the treatment of tumors, mainly of the oropharynx and larynx. The use of robotic surgery improves visualization of the operative field due to its three-dimensional image and enhances the surgeon’s dexterity due to bimanual control of the robotic arms. Furthermore, the assistant contributes with suction and tissue traction, which leads to the use of four instruments during surgery, something impossible during a transoral resection through laryngoscopy, for instance. Therefore,

Locally Advanced Laryngeal Cancer

Program Description: This miniseminar is designed to update general otolaryngologists on cutting edge advances in advanced laryngeal cancer. Educational Objectives: 1) Improve knowledge of the treatment principles of advanced laryngeal cancer. 2) Surgical organ preservation treatment for advanced laryngeal cancer, including robotic surgery. 3) Post-treatment functional outcomes.

CCR2-64I polymorphism and head and neck squamous cell carcinoma

OBJECTIVES: From the differential expression profiles between paired nasopharyngeal carcinoma (NPC) and pericancerous normal epithelium analyzed by cDNA microarray technology, we have found that CCL20 might be a potential biomarker. Since CCL20 is a secretory protein, we herein further examine whether it could be used as a serum marker for NPC detection and correlated with prognosis. METHODS: The study enrolled a prospective and a retrospective cohort, which totally comprises 275 NPC patients and 250 controls. CCL20 levels in sera were measured by ELISA. EBV DNA load and EBV VCA IgA were measured by qRT-PCR and immunofluorescent assay, respectively. RESULTS: Serum CCL20 levels were significantly higher in untreated patients, recurrent patients and patients with distant metastases versus non-NPC controls, patients with complete remission, and long-term disease-free patients. Serum CCL20 levels were significantly higher in untreated NPC patients with advanced TNM stage. Measurement of CCL20, EBV DNA and VCA IgA levels in serial serum/plasma samples from treated patients at six-month intervals revealed a high association between CCL20 serum level and disease status. Among 155 consecutive NPC patients, subjects with pre-treated CCL20 serum levels over 65 pg/ml had worse prognoses for overall survival and distant metastasis-free survival in univariate and multivariate analysis. CONCLUSIONS: In this study, we found that serum CCL20 levels were positively correlated with the current state-of-art NPC marker, EBV DNA load, implying its potential correlated with NPC tumor burden. CCL20 might be a novel serum marker for NPC detection and prediction of treatment outcome.