Yu-Liang Chang

Complementary serum test of antibodies to Epstein-Barr virus nuclear antigen-1 and early antigen: a possible alternative for primary screening of nasopharyngeal carcinoma.

This hospital-based cohort study evaluated the efficacy of three Epstein-Barr virus (EBV) - associated assays for nasopharyngeal carcinoma (NPC) primary screening and monitoring treatment outcome. Five hundred and seventeen consecutive subjects, including 156 NPC patients, 264 healthy volunteers and 97 patients with head and neck squamous cell carcinoma (HNSCC) were enrolled. The sensitivity and specificity of EBV IgAs to viral capsid antigen (VCA), complementary EBV IgAs to early antigen and nuclear antigen-1 (EA+EBNA-1), and EBV DNA load were examined by immunofluorescent assays, enzyme-linked immunosorbent assays, and quantitative real-time PCR, respectively. After constructing the receiver operating characteristics to demonstrate screening efficacy, EBV EA+EBNA-1 IgA (AUC: 0.952; 95% CI, 0.930-0.974) was proved superior to EBV VCA IgA (AUC: 0.888; 95% CI, 0.854-0.922) or EBV DNA load (AUC: 0.893; 95% CI, 0.854-0.932) in differentiating NPC patients from controls. Comparison of screening efficacy between NPC patients and HNSCC patients revealed EBV EA+EBNA-1 IgA (AUC: 0.964; 95% CI, 0.943-0.985) still outperformed EBV VCA IgA (AUC: 0.884; 95% CI, 0.845-0.923). In subjects with higher serum titer or level equal to or above 1:80 and 6 EU/ml for EBV VCA IgA and EA+EBNA-1 IgA, the specificity reached as high as 99.2% and 95.1%, respectively, in the control groups. However, correlation of these three assays with clinicopathological manifestations of NPC, revealed only EBV DNA load significantly associated with N stage and overall stage in NPC patients. Additionally, EBV DNA load could be used to further raise the specificity of EBV EA+EBNA-1 IgA assays and was also the only assay to be consistently predictive of tumor relapse in post-treatment patients according to serial test results by time frame. Consequently, an EBV EA+EBNA-1 IgA-based protocol is recommended for mass screening, but EBV DNA load should be used solely for post-treatment monitoring for NPC in endemic areas.

Overexpression of activin A in oral squamous cell carcinoma: association with poor prognosis and tumor progression.

BackgroundBoth activin A, a member of transforming growth factor β superfamily, and its inhibitor follistatin have been shown to be overexpressed in various cancers. We examined the potential role of activin A and follistatin in tissue and blood samples from patients with oral squamous cell carcinoma.MethodsFor activin A and follistatin, the expression of tissue samples from 92 patients was examined by immunohistochemical study, and the serum levels of blood samples from 111 patients and 91 healthy controls were measured by enzyme-linked immunosorbent assay.ResultsWe found that overexpression of immunohistochemically detected activin A was correlated with positive N stage, poor histological differentiation, and perineural invasion (P = 0.029, 0.002, and 0.014, respectively). In survival analyses, patients with oral squamous cell carcinoma, whose tumors overexpressed activin A, had a worse prognosis for overall survival and disease-free survival (P = 0.009 and 0.007). However, expression of follistatin in tumor was not correlated with overall survival or disease-free survival. Serum activin A and follistatin levels in 111 untreated patients were neither significantly different from those of 91 control samples nor associated with any clinicopathological manifestations. In vitro suppression of activin A expression in OC3 cells using specific interfering RNA-attenuated cell proliferation, migration, and invasiveness.ConclusionsThese findings suggest that activin A overexpression in oral squamous cell carcinomas is associated with patients’ survival and may contribute to tumor progression and metastasis.

Prognostic cytokine markers in peripheral blood for oral cavity squamous cell carcinoma identified by multiplexed immunobead-based profiling.

BACKGROUND Oral cavity squamous cell carcinoma (OSCC) is one of the most common cancers worldwide and has been considered to be highly associated with altered biological processes, including immunocyte chemotaxis, inflammatory response, angiogenesis, and/or immune regulation, suggesting that the levels of the tumor-related cytokines and chemokines will be dysregulated in the tumor microenvironment as well as in the systemic circulation and might be associated with some OSCC phenotypes. METHODS To profile cytokines in OSCC patients, the plasma levels of 48 proteins (26 cytokines, 10 chemokines, and 12 growth factors) were measured in 111 untreated OSCC patients, 112 healthy individuals, and 107 individuals with oral premalignant lesion (OPL). RESULT Compared to the plasma levels in the healthy individuals and OPL group, the levels of 12 proteins were significantly dysregulated in the OSCC patients. Further analysis demonstrated that the levels of IFN-α2, IL-2RA, and SCF were significantly lower in patients with higher pT status. IFN-α2 levels also decreased in patients with higher tumor depths. Moreover, OSCC patients with greater levels of VEGF (>4.87 pg/ml) before treatment had worse prognoses for overall survival after treatment (P=0.035). CONCLUSIONS This is the first report showing that the plasma VEGF levels may be a useful prognostic indicator of OSCC.

Overexpression of macrophage inflammatory protein-3α in oral cavity squamous cell carcinoma is associated with nodal metastasis

We examined the role of macrophage inflammatory protein (MIP)-3α on oral cavity squamous cell carcinoma (OSCC) and whether it was involved in modulating OSCC cell functions. The study population was comprised of 102 patients with OSCC. MIP-3α levels in tissues were examined by immunohistochemistry and quantitative real-time RT-PCR. Effects of MIP-3α on OSCC cell function were investigated by cell proliferation assays, trans-well migration/invasion assays, and RNA interference. We found that MIP-3α was overexpressed in OSCC tumor cells. MIP-3α expression was significantly higher in tumor cells vs. normal epithelial cells, as determined by both quantitative real-time RT-PCR and immunohistochemistry. Overexpression of MIP-3α was significantly correlated with positive pN status (P=0.036). Nevertheless, there were no correlations related to patient age, pT status, overall pathological stage, cell differentiation, or perineural invasion. The long-term disease-specific survival for patient subgroups stratified by the absence or presence of MIP-3α overexpression was 70.9% vs. 54.7% (P=0.041). Multivariate analysis indicated that MIP-3α overexpression had a significantly lower disease-specific survival (hazard ratio: 2.158; P=0.037). Additionally, in vitro suppression of MIP-3α expression in OECM-1 cells using specific interfering RNAs attenuated cell migration and invasiveness. These findings suggest that MIP-3α overexpression in OSCC is associated with a poorer prognosis for patient survival and contributes to tumor metastasis.

Serum levels of chemokine (C-X-C motif) ligand 9 (CXCL9) are associated with tumor progression and treatment outcome in patients with oral cavity squamous cell carcinoma

OBJECTIVES The aim of this cohort study was to examine the role of chemokine (C-X-C motif) ligand 9 (CXCL9) on oral cavity squamous cell carcinoma (OSCC). METHODS Sera from 181 OSCC patients, 231 healthy individuals, and 50 OSCC tumor samples were enrolled. CXCL9 expression in tissue samples was analyzed by quantitative real-time PCR and immunohistochemistry. CXCL9 serum concentrations were measured by enzyme-linked immunosorbent assay. Effects of CXCL9 on OSCC cell function were investigated by cell proliferation assays, trans-well migration/invasion assays, and RNA interference. RESULTS CXCL9 expression was significantly higher than for normal epithelium in the tissue samples. CXCL9 serum concentrations were also significantly higher in OSCC patients compared to those in healthy individuals. Serum CXCL9 levels were significantly higher in OSCC patients with higher pT status, pathological overall stages, tumor depths, and positive bone invasion (P = 0.033, 0.004, 0.041, and 0.002, respectively). Moreover, OSCC patients with higher CXCL9 levels (> 209 pg/mL, median level) before treatment had worse prognoses for overall survival and disease-specific survival (P = 0.0006 and 0.0009, respectively). Multivariate logistic regression analyses also indicated that higher CXCL9 serum levels were an independent prognostic factor for overall survival and disease-free survival (P = 0.003 and 0.004, respectively). The in vitro suppression of CXCL9 expression in SCC25 cells using specific interfering RNAs attenuated cell proliferation, migration and invasiveness. CONCLUSIONS Our study demonstrated that CXCL9 is associated with tumor burden and aggressiveness of OSCC tumors and serum level of this ligand may be useful as a prognostic indicator.

Evaluation of Lymphatic and Vascular Invasion in Relation to Clinicopathological Factors and Treatment Outcome in Oral Cavity Squamous Cell Carcinoma

AbstractThis study evaluated the associations between lymphatic and vascular invasion of oral cavity squamous cell carcinoma (OSCC) and clinicopathological manifestations, as well as their impact on patient outcomes after treatment.In total, 571 patients with primary OSCC who underwent surgery with or without adjuvant therapy were enrolled.Lymphatic and vascular invasion were found in 28 (5%) and 16 (3%) patients, respectively. Significant associations were found between lymphatic and vascular invasion and overall stage (P < 0.001 and P = 0.020, respectively), tumor stage (P = 0.009 and P = 0.025, respectively), nodal metastasis (both P < 0.001), extracapsular spread (both P < 0.001), perineural invasion (both P < 0.001), bone invasion (P = 0.004 and P = 0.001, respectively), depth of invasion (P < 0.001 and P = 0.001, respectively), and pathologic differentiation (P = 0.002 and P < 0.001, respectively). In the analysis of adverse events during follow-up, neither lymphatic nor vascular invasion was statistically associated with local recurrence, neck recurrence, and distant metastasis. Although lymphatic invasion exhibited significant associations with poorer overall survival (P < 0.001), disease-specific survival (P < 0.001), and disease-free survival (P = 0.01), it was not demonstrated to be an independent prognostic factor in all multivariate analyses.Although both lymphatic and vascular invasion are associated with many clinicopathological manifestations, neither affects the occurrence of locoregional recurrence and distant metastasis in patients with OSCC after treatment.

Pretreatment Interleukin-6 Serum Levels Are Associated with Patient Survival for Oral Cavity Squamous Cell Carcinoma

Objective This study aims to determine the role of serum interleukin-6 concentration for oral cavity squamous cell carcinomas. Study Design Cohort study. Setting Tertiary referral center. Methods Two hundred thirty-seven untreated patients, 125 healthy individuals, and 104 individuals with oral premalignant lesions were enrolled. Interleukin-6 serum concentrations were measured by enzyme-linked immunosorbent assay. Results Serum concentrations of interleukin-6 were significantly higher in patients compared with the levels in healthy individuals and the subjects with oral premalignant lesions. Serum interleukin-6 levels were significantly higher in patients with higher pT status (from pT1 to pT4, median values in pg/mL = 0, 0, 1.3, and 5.0, respectively, with P < .001), higher pathological stages (from stage I to IV, median values = 0, 0, 1.3, and 3.6, respectively, with P < .001), positive bone invasion (5.0 vs 0, 1.4 vs 0; P < .001), and higher tumor depths (1.4 vs 0; P = .005). Patients with higher pretreatment levels of interleukin-6 (>1.35 pg/mL, median level) had worse prognoses for 5-year overall survival and disease-specific survival despite treatment (75.7% vs 54.9% and 79.1% vs 59.8%; P = .001 and .003, respectively). Multivariate logistic regression analyses also indicated that higher interleukin-6 serum levels were an independent prognostic factor for overall survival and disease-free survival (adjusted hazard ratio = 2.417 and 2.364; P = .009 and .017, respectively). Conclusion Our study revealed that serum interleukin-6 levels were associated with increased tumor burden and aggressiveness of oral cavity squamous cell carcinomas and may be useful as a prognostic indicator after treatment.

Oral Microbiota Community Dynamics Associated With Oral Squamous Cell Carcinoma Staging

Oral squamous cell carcinoma (OSCC) is a highly aggressive cancer and the fourth leading malignancy among males in Taiwan. Some pathogenic bacteria are associated with periodontitis and oral cancer. However, the comprehensive profile of the oral microbiome during the cancers progression from the early stage to the late stage is still unclear. We profiled the oral microbiota and identified bacteria biomarkers associated with OSCC. The microbiota of an oral rinse from 51 healthy individuals and 197 OSCC patients at different stages were investigated using 16S rRNA V3V4 amplicon sequencing, followed by bioinformatics and statistical analyses. The oral microbiota communities from stage 4 patients showed significantly higher complexity than those from healthy controls. The populations also dynamically changed with the cancers progression from stage 1 to stage 4. The predominant phyla in the oral samples showed variation in the relative abundance of Fusobacteria, Bacteroidetes, and Actinobacteria. The abundance of Fusobacteria increased significantly with the progression of oral cancer from the healthy controls (2.98%) to OSCC stage 1 (4.35%) through stage 4 (7.92%). At the genus level, the abundance of Fusobacterium increased, while the number of Streptococcus, Haemophilus, Porphyromonas, and Actinomyces decreased with cancer progression. Fusobacterium periodonticum, Parvimonas micra, Streptococcus constellatus, Haemophilus influenza, and Filifactor alocis were associated with OSCC, and they progressively increased in abundance from stage 1 to stage 4. The abundances of Streptococcus mitis, Haemophilus parainfluenzae, and Porphyromonas pasteri were inversely associated with OSCC progression. We selected a bacterial marker panel of three bacteria (upregulated F. periodonticum, down-regulated S. mitis, and P. pasteri), which had an AUC of 0.956 (95% CI = 0.925–0.986) in discriminating OSCC stage 4 from the healthy controls. Furthermore, the functional prediction of oral bacterial communities showed that genes involved in carbohydrate-related metabolism, such as methane metabolism, and energy-metabolism-related parameters, such as oxidative phosphorylation and carbon fixation in photosynthetic organisms, were enriched in late-stage OSCC, while those responsible for amino acid metabolism, such as folate biosynthesis and valine, leucine, and isoleucine biosynthesis, were significantly associated with the healthy controls. In conclusion, our results provided evidence of oral bacteria community changes during oral cancer progression and suggested the possibility of using bacteria as OSCC diagnostic markers.

Nomogram based on albumin and neutrophil-to-lymphocyte ratio for predicting the prognosis of patients with oral cavity squamous cell carcinoma

Increasing evidence indicates that inflammation plays a crucial role in cancer development. A novel scoring system based on albumin and the neutrophil-to-lymphocyte ratio (NLR) was developed and incorporated into a nomogram to create a more accurate prognostic tool for oral cavity squamous cell carcinoma (OSCC) patients. A retrospective review was performed on 613 consecutive patients undergoing ablative surgery for OSCC between September 2005 and December 2014. NLR and albumin were determined and used to calculate an albumin/NLR score (ANS). The nomogram was based on the ANS and several clinicopathological manifestations, and its accuracy was determined by the concordance index (c-index). A high ANS was significantly associated with aggressive tumor behaviors, such as T status, overall stage, extranodal extension, perineural invasion, tumor depth, and decreased overall survival (OS). Multivariate analysis indicated that age, overall stage, extranodal extension, and ANS were independent factors for OS. The c-index for OS prognosis was 0.750 using this nomogram compared to 0.688 using TNM staging alone. The prognostic accuracy for OS in OSCC patients can be significantly improved using a nomogram that incorporates the novel ANS and other clinicopathological variables.

Clinical Implications of Tumor-Associated Tissue Eosinophilia in Tongue Squamous Cell Carcinoma: Implications of TATE in Tongue SCC

The role of tumor‐associated tissue eosinophilia (TATE) in oral cavity cancer remains quite controversial. This study investigated the potential role of TATE in tongue squamous cell carcinoma (TSCC).