Fascin is a circulating tumor marker for head and neck cancer as determined by a proteomic analysis of interstitial fluid from the tumor microenvironment.

BACKGROUND: Head and neck cancer (HNC) is a prevalent cancer worldwide; however, clinically useful tumor markers for HNC have not been identified. Here, we aimed to identify secretory proteins from the tumor microenvironment as candidate circulating tumor markers. METHODS: Samples derived from seven pairs of tumor interstitial fluid (TIF) and normal interstitial fluid (NIF) samples from patients with HNC were analyzed. The proteomes were determined by gel-based-mass-spectrometry proteomic methods. The most up-regulated protein, fascin was confirmed in the cancer tissues and cell culture supernatant by immunoblotting and immunohistochemistry assays. Serum fascin was determined in 40 HNC and 40 normal individuals by ELISA. RESULTS: After proteomics analysis, 189 peptides were identified, corresponding to 75 proteins. Of the 21 proteins which were identified more than twice, five up-regulated proteins identified most frequently including fascin. The most elevated fascin was over-expressed in cancer tissues and cell culture supernatant. Serum fascin was significantly up-regulated in the cancer patients (p<0.001) and correlated with pathological lymph node metastasis (p=0.022). To assess the diagnostic efficacy, serum levels of fascin and another potential biomarker SCCA were determined. Fascin showed a high predictable value with an area under the curve (AUC) of 0.808 (95% CI 0.723-0.901) in the receiver operator curve (ROC), compared to 0.501 (95% CI 0.378-0.634) for SCCA. CONCLUSIONS: We have identified 75 potential circulating tumor markers associated with HNC, including fascin. Serum fascin could discriminate cancer patients from healthy individuals; thus, it may serve as a circulating biomarker for HNC.

Combined determination of circulating miR-196a and miR-196b levels produces high sensitivity and specificity for early detection of oral cancer.

OBJECTIVES: The aim of this study was to determine whether the oncogenic microRNA family members miR-196a and miR-196b can be circulating biomarkers for the early detection of oral cancer. DESIGN AND METHODS: To determine the stability of circulating miRNA, the blood sample was aliquot and stored at different temperature conditions for analysis. To assess the diagnostic efficacy, we determined the levels of miR-196s in plasma samples, including 53 from healthy individuals, 16 from pre-cancer patients, and 90 from oral cancer patients. RESULTS: In general, circulating miRNA was very stable when storing plasma samples at -20°C or below. In clinical study, both circulating miR-196a and miR-196b were substantially up-regulated in patients with oral pre-cancer lesions (5.9- and 14.8-fold, respectively; P < 0.01), as well as in oral cancer patients (9.3- and 17.0-fold, respectively; P < 0.01). These results show prominent discrimination between normal and pre-cancer patients (AUC = 0.764 or 0.840, miR-196a or miR-196b, respectively), and between normal and cancer patients (AUC = 0.864 or 0.960, miR-196a or miR-196b, respectively). The combined determination of miR-196a and miR-196b levels produces excellent sensitivity and specificity in the diagnosis of patients with oral pre-cancer (AUC = 0.845) or oral cancer (AUC = 0.963), as well as in the prediction of potential malignancy (AUC = 0.950, sensitivity = 91%, specificity = 85%). CONCLUSION: Combined determination of circulating miR-196a and miR-196b levels may serve as panel plasma biomarkers for the early detection of oral cancer.

OncomiR-196 promotes an invasive phenotype in oral cancer through the NME4-JNK-TIMP1-MMP signaling pathway.

BACKGROUND: MicroRNA-196 (miR-196), which is highly up-regulated in oral cancer cells, has been reported to be aberrantly expressed in several cancers; however, the significance of miR-196 in oral cancer has not yet been addressed. METHODS: Cellular functions in response to miR-196 modulation were examined, including cell growth, migration, invasion and radio/chemosensitivity. Algorithm-based studies were used to identify the regulatory target of miR-196. The miR-196 target gene and downstream molecular mechanisms were confirmed by RT-qPCR, western blot, luciferase reporter and confocal microscopy analyses. miR-196 expression was determined in paired cancer and adjacent normal tissues from oral cancer patients. RESULTS: Both miR-196a and miR-196b were highly over-expressed in the cancer tissue and correlated with lymph node metastasis (P = 0.001 and P = 0.006, respectively). Functionally, miR-196 actively promoted cell migration and invasion without affecting cell growth. Mechanistically, miR-196 performed it's their function by inhibiting NME4 expression and further activating p-JNK, suppressing TIMP1, and augmenting MMP1/9. CONCLUSION: miR-196 contributes to oral cancer by promoting cell migration and invasion. Clinically, miR-196a/b was significantly over-expressed in the cancer tissues and correlated with lymph node metastasis. Thus, our findings provide new knowledge of the underlying mechanism of cancer metastasis. miR-196 may serve as a promising marker for better oral cancer management.

Free flap reconstruction after surgical release of oral submucous fibrosis: long-term maintenance and its clinical implications.

BACKGROUND AND AIM: Oral submucous fibrosis (OSF) is an insidious disease with progressive limitation of mouth opening and potential malignant change of the oral mucosa. Cancer surveillance is of utmost importance, but it is often limited by severe trismus. Surgical release and free flap reconstruction is effective but its long-term efficacy has not been completely established. This work aims to review our experience in the past 15 years in surgical release of OSF-related trismus followed by free flap reconstruction. METHODS: Patient's age, gender, smoking history, drinking history and betel-nut consumption history were retrieved. Surgical release and reconstructive procedures were detailed. Inter-incisor distances (IIDs) were measured preoperatively (PO-IID), intra-operatively after maximal release (IO-IID) and during the last follow-up (FU-IID). Subsequent development of oral cancers (oral squamous cell carcinoma, OSCC) and relevant details were documented. Potential predictors of long-term IID gain were analysed. RESULTS: A total of 92 patients were included in our study. There was a significant difference (p = 0.000) in PO-IID (13.8 ± 6.6 mm) and FU-IID (27.2 ± 8.8 mm) indicating the long-term efficacy of the release procedure. The mean long-term IID gain was 13.0 ± 7.5 mm. Bilateral coronoidectomy resulted in a greater degree of intra-operative gain in IID (p = 0.025). PO-IID (r = -0.277, p = 0.001) and intra-operative gain in IID (r = 0.198, p = 0.001) were found to be predictive of long-term IID gain. Ten patients (11%) developed OSCC during our study period. CONCLUSIONS: Aggressive surgical release (with bilateral coronoidectomy if necessary) followed by free flap reconstruction is an effective treatment for OSF-related trismus. Our study has confirmed its long-term efficacy and its important role in cancer surveillance.

Oncogenic function and early detection potential of miRNA-10b in oral cancer as identified by microRNA profiling.

The miRNA participates in a variety of biologic processes, and dysregulation of miRNA is associated with malignant transformation. In this study, we determined specific profile of miRNA associated with oral cancer by using miRNA array screening method. There were 23 miRNAs found with considerably differential expressions between six oral cancer cell lines and five lines of normal oral keratinocytes, in which, 10 miRNAs showed the highest significant difference after independent examination by reverse transcription quantitative PCR. Eight molecules were upregulated, miR-10b, miR-196a, miR-196b, miR-582-5p, miR-15b, miR-301, miR-148b, and miR-128a, and two molecules, miR-503 and miR-31, were downregulated. The most upregulated miR-10b was further examined, and its functions were characterized in two oral cancer cell lines. The miR-10b actively promotes cell migration (2.6- to 3.6-fold) and invasion (1.7- to 1.9-fold) but has minimal effect on cell growth or chemo-/radiosensitivity. Furthermore, miR-10b was considerably elevated in the plasma of xenografted tumor mice (20-fold). This upregulation of miR-10b in plasma was further shown in the patients with oral cancer [P < 0.0001, area under curve (AUC) = 0.932] and precancer lesions (P < 0.0001, AUC = 0.967), suggesting that miR-10b possesses a high potential to discriminate the normal subjects. In conclusion, we have identified at least 10 miRNAs significantly associated with oral cancer, including the most elevated miR-10b. The miR-10b actively participates in cancer formation by promoting cell migration and invasion. Our study using clinical samples suggests that plasma miR-10b has high potential as an early detection marker for oral cancer.

Caldwell-Luc operation without inferior meatal antrostomy: a retrospective study of 50 cases.

PURPOSE: In the standard Caldwell-Luc operation, an inferior meatal antrostomy is performed to promote sinus drainage. However, inferior meatal antrostomy has been criticized for its additional operation time and wound, early loss of the opening, and risk of injury to the nasolacrimal duct. This study retrospectively reviewed the results of the Caldwell-Luc operation without inferior meatal antrostomy in the treatment of odontogenic maxillary sinusitis or odontogenic sinus disease. PATIENTS AND METHODS: The records of 50 patients who had an odontogenic sinus disease and underwent the Caldwell-Luc operation without inferior meatal antrostomy were reviewed. The data included the patient's age, gender, surgical indications, surgical condition, and complications. RESULTS: From April 2004 to October 2010, there were 27 male patients and 23 female patients aged 14 to 70 years (mean, 37 years) who underwent the modified Caldwell-Luc operation. The surgical indications included intrasinus odontogenic cysts (44%), oroantral fistulae with chronic sinusitis (44%), odontoma (4%), odontogenic sinusitis (4%), and foreign bodies in the maxillary sinus (4%). The patients were successfully treated with minimal complications. CONCLUSIONS: The modified Caldwell-Luc operation provides easier postoperative care and involves fewer complications. It is not necessary to create the inferior meatal antrostomy in the Caldwell-Luc operation when treating odontogenic sinus disease.

Positive association of glucose-regulated protein 78 during oral cancer progression and the prognostic value in oral precancerous lesions.

BACKGROUND: Our aim was to examine the potential role of glucose-regulated protein (GRP)78 during oral cancer progression and the prognostic value in oral precancerous lesions. METHODS: A total of 204 patients with oral cancer and 86 with precancerous lesions were investigated. GRP78 expression was determined in the lesion tissues by Western blot analysis. Association of GRP78 with clinicopathology or disease prognosis was examined using Fisher's exact, Kaplan-Meier, or Cox regression method. RESULTS: Hyperexpression of GRP78 was found to be correlated with increasing malignant potential of oral lesions, with 14% in leukoplakia, 27% in erythroplakia, 50% in verrucous lesion, and 74% in oral cancer (p < .0001), suggesting this molecule plays a crucial role in the early steps of oral oncogenesis. In patients with precancerous lesions of the oral cavity, GRP78 expression predicts poorer same-site premalignancy-free survival (p = .002) and malignancy-free survival rates (p = .002). CONCLUSION: Determination of GRP78 expression levels might enable a better risk stratification for patients with oral premalignant lesions.

T4b oral cavity cancer below the mandibular notch is resectable with a favorable outcome.

According to the AJCC 2002 staging system, squamous cell carcinoma of the oral cavity (OSCC) with T4b is unresectable. Herein, we report on our surgical outcome for T4b OSCC. All patients were enrolled before 2002. From January 1996 to December 2000, 45 consecutive untreated T4b OSCC patients were included. According to the trans-axial plane of the mandibular notch on CT/MRI, seven were supra-notch T4b and 38 were infra-notch T4b tumors. Significantly higher 5-year loco-regional control and survivals were observed in patients with infra-notch T4b than with supra-notch T4b. In the infra-notch group, pN0-1 patients had a significantly higher 5-year loco-regional control and survivals than pN2 patients. Nerve invasion was the sole significant adverse factor for loco-regional control and survivals in the infra-notch T4b group with pN0-1. Our retrospective study demonstrated that OSCC patients of infra-notch T4b with pN0-1 and no nerve invasion might have a favorable surgical outcome.

Identification of differentially expressed genes in oral squamous cell carcinoma (OSCC): overexpression of NPM, CDK1 and NDRG1 and underexpression of CHES1.

To identify cellular genes that could potentially serve as predictive molecular markers for human oral cancer, we employed differential display analysis to compare the gene expression profiles between oral squamous cell carcinoma (OSCC) and histopathologically normal epithelium tissues. Comparative real-time RT-PCR was used to confirm the gene expression in 52 OSCC patients, and a 2-fold difference was defined as over- or underexpression. A total of 7 genes were identified: NPM, CDK1, NDRG1, HMGCR, EF1A, NAC and CHES1. In the cancer tissues, NPM, CDK1 and NDRG1 were significantly overexpressed (an average of 7.6-, 17.2- and 12.9-fold, respectively), and CHES1 was underexpressed (15-fold). The frequencies of the differential expression were 40, 56, 67 and 46%, respectively in NPM, CDK1, NDRG1 and CHES1. In Western blot analysis, the protein expressions of NPM, CDK1 and NDRG1 were also increased in the cancer tissues, consistent with the mRNA expression results. To further evaluate clinicopathological associations in these genes, Pearson chi-square analysis was employed. Levels of CDK1 and NDRG1 were associated with poorly differentiated tumors (p = 0.043 and 0.023), suggesting that these genes participate in the mechanism of tumor transformation. Expressions of CDK1 and NDRG1, and CDK1 and CHES1 were mutually statistically correlated (p = 0.001 and 0.014), indicating that these genes share a very close regulatory relationship or interact synergistically in oncogenesis. In conclusion, we identified 7 genes that are differentially expressed in OSCC, and we provide the first evidence that NPM, CDK1 and NDRG1 are overexpressed and CHES1 is underexpressed in oral cancer. These results serve as a fundamental base for employing these genes in future clinical applications.