Serpin peptidase inhibitor, clade E, member 2 is associated with malignant progression and clinical prognosis in oral squamous cell carcinoma.

Background/purpose: The serpin peptidase inhibitor, clade E, member 2 (SERPINE2), is upregulated in breast cancer, prostate cancer, and urothelial carcinoma; however, limited information exists regarding its expression in oral cancer. Therefore, this study aimed to analyze the association between SERPINE2 expression and oral squamous cell carcinoma (OSCC) outcomes. Materials and methods: SERPINE2 mRNA and protein expression in patients with head and neck squamous cell carcinoma and OSCC were investigated using online databases and tissue-array analysis. Its relationship with clinicopathological characteristics, OSCC prognosis and its biological function in OSCC cells were explored. Results: Analysis using online databases revealed higher SERPINE2 expression in tumor tissues and its role as a prognostic factor. High SERPINE2 protein levels were significantly correlated with adverse pathological parameters, including advanced clinical stage and tumor status (P < 0.001), lymph nodes (P = 0.014), and distant metastases (P = 0.013). High SERPINE2 expression was associated with worse overall survival (P < 0.001) and was identified as an independent prognostic factor for OSCC. In vitro studies revealed that SERPINE2 knockdown significantly reduced cell proliferation, migration, and invasion in OSCC cell lines. Conclusion: This study suggests that SERPINE2 may serve as a prognostic biomarker and potential therapeutic target for oral cancer.

Identification of Somatic Mutations in Plasma Cell-Free DNA from Patients with Metastatic Oral Squamous Cell Carcinoma.

The accurate diagnosis and treatment of oral squamous cell carcinoma (OSCC) requires an understanding of its genomic alterations. Liquid biopsies, especially cell-free DNA (cfDNA) analysis, are a minimally invasive technique used for genomic profiling. We conducted comprehensive whole-exome sequencing (WES) of 50 paired OSCC cell-free plasma with whole blood samples using multiple mutation calling pipelines and filtering criteria. Integrative Genomics Viewer (IGV) was used to validate somatic mutations. Mutation burden and mutant genes were correlated to clinico-pathological parameters. The plasma mutation burden of cfDNA was significantly associated with clinical staging and distant metastasis status. The genes TTN, PLEC, SYNE1, and USH2A were most frequently mutated in OSCC, and known driver genes, including KMT2D, LRP1B, TRRAP, and FLNA, were also significantly and frequently mutated. Additionally, the novel mutated genes CCDC168, HMCN2, STARD9, and CRAMP1 were significantly and frequently present in patients with OSCC. The mutated genes most frequently found in patients with metastatic OSCC were RORC, SLC49A3, and NUMBL. Further analysis revealed that branched-chain amino acid (BCAA) catabolism, extracellular matrix-receptor interaction, and the hypoxia-related pathway were associated with OSCC prognosis. Choline metabolism in cancer, O-glycan biosynthesis, and protein processing in the endoplasmic reticulum pathway were associated with distant metastatic status. About 20% of tumors carried at least one aberrant event in BCAA catabolism signaling that could possibly be targeted by an approved therapeutic agent. We identified molecular-level OSCC that were correlated with etiology and prognosis while defining the landscape of major altered events of the OSCC plasma genome. These findings will be useful in the design of clinical trials for targeted therapies and the stratification of patients with OSCC according to therapeutic efficacy.

Exploiting salivary miR-375 as a clinical biomarker of oral potentially malignant disorder.

Background/purpose: Oral potentially malignant disorder (OPMD) is an important premalignancy worldwide. MicroRNAs (miRNAs) are endogenously expressed non-coding RNAs that regulate the post-transcriptional levels of targeted mRNAs. MiRNA-375 (miR-375) is markedly downregulated in oral carcinoma tissues and plays an oncogenic role in oral carcinogenesis. We explored the potential of the deregulated salivary miR-375 levels in OPMD patients. Materials and methods: . We analyzed the levels of miR-375 in the saliva of patients with OPMD (n = 45) and healthy controls (n = 24) by quantitative RT-PCR. The cell lysates and supernatants were treated with the miR-375 mimic and inhibitor. Results: Salivary miR-375 levels were decreased markedly in the patients with OPMD, compared with the controls. OPMD patients with non-dysplasia showed a higher abundance of miR-375 in the saliva than dysplasia patients, suggesting that salivary miR-375 is a more sensitive marker for OPMD. Patients with malignant transformation during the follow-up period showed lower expression of saliva miR-375 than the others. MiR-375 expression was markedly decreased by treatment with the miR-375 inhibitor, and the supernatants of both NHOK and SAS cells showed a corresponding decline in miR-375 expression. Conclusion: Our results indicate the potential application of salivary miR-375 as a biomarker for the detection and long-term follow-up of OPMD.

Risk factors for reconstruction plate exposure after surgery in oral cancer patients. A retrospective cohort study.

OBJECTIVES: The aim of this study was to elaborate risk factors for reconstruction plate exposure after wide excision in oral cancer patients, and to find out the most effective treatment. MATERIALS AND METHODS: We include patients who underwent ablative surgery for oral cancer and reconstruction plate fixation from the year 2010 to 2016, separate them into two groups according to whether the hardware was exposed, compare risk factors including age, tumor site, staging, comorbidities, and previous treatment between the two groups. The treatment course and outcome were also recorded. RESULTS: In total, 75 patients received reconstruction plate fixation after ablative surgery. Bone plate exposure was found in 26 cases (34.6%). The size of the bone defect and the thickness of soft tissue covering the plate were significant risk factors for plate exposure. In 21 patients (72.4%), the bone plate was removed. Conservative treatment was not effective. Removal of bone plate and debridement in one single surgery had a success rate of 81%. CONCLUSION: In this study, skin thickness less than 1.5 mm over the reconstruction plate and bone defect size larger than 8.4 cm were the two significant risk factors for bone plate exposure. Although a standardized treatment algorithm is lacking, surgical debridement with removal of the bone plate result in complete soft tissue healing in most patients and should be the treatment of choice. CLINICAL RELEVANCE: Patients with larger bone defect and thinner covering soft tissue bear more risk for exposure. The most effective treatment is to remove the hardware.

Phlebosclerotic colitis: an analysis of clinical and CT findings in 29 patients with long-term follow-up.

BACKGROUND: Phlebosclerotic colitis (PC) is a rare form of nonthrombotic colonic ischemia. This retrospective study analyzed the clinical findings and temporal CT changes in 29 PC patients with long-term follow-up. METHODS: Twenty-nine patients with characteristic CT features of PC collected between 1997 and 2020 were stratified into the acute abdomen group (AA-group) (n = 10), chronic-progressive group (CP-group) (n = 14) and chronic-stable group (CS-group) (n = 5). Clinical and CT changes during follow-up, comorbidities and final outcomes were compared. RESULTS: The AA-group exhibited a significantly thicker colonic wall and more involved segments and pericolic inflammation than the CP-group and CS-group on initial CT (p = < 0.001-0.031). Seven patients in the AA-group who underwent right hemicolectomy had no recurrence during follow-up (mean ± SD, 7.1 ± 3.3 years), and the remaining three patients with renal or hepatic comorbidities who underwent conservative treatment died within 14 days. The CP-group showed significantly higher frequencies of chronic renal failure, urinary tract malignancies and liver cirrhosis than the AA-group (p = 0.005-0.008). In addition, CT follow-up (7.9 ± 4.3 years) showed significant increases in mesenteric venous calcifications, colonic wall thickening and involved colonic segments (p = 0.001-0.008) but conservative treatments were effective. The CS-group remained unchanged for years (8.2 ± 3.9 years). CONCLUSIONS: Early surgery offered excellent prognosis in PC-related acute abdomen denoted by marked right colonic wall thickening and pericolic inflammation on CT. Conservative treatments with a wait-and-watch strategy were appropriate for CP-PC and CS-PC, albeit CP-PC harbored significant increases in calcifications, colonic wall thickening and affected segments in long-term CT follow-up.

Droplet digital polymerase chain reaction for detection and quantification of cell-free DNA TP53 target somatic mutations in oral cancer.

BACKGROUND: TP53 mutation is a driver mutation of oral carcinogenesis. This study investigated cancerous and cell-free DNA (cfDNA) in patients with oral squamous cell carcinoma (OSCC) to detect the target hotspot somatic mutation of TP53. OBJECTIVE: TP53 target hotspot mutations were determined in surgically resected primary tumor samples from 107 OSCC patients. METHODS: Cancerous and cfDNA samples were examined for mutations through droplet digital polymerase chain reaction (ddPCR) by using mutation-specific assays. The ddPCR results were evaluated alongside clinicopathological data. RESULTS: In total, 23 cases had target TP53 mutations in varying degrees. We found that OSCC had relatively low cfDNA shedding, and mutations were at low allele frequencies. Of these 23 cases, 13 had target TP53 mutations in their corresponding cfDNA. Target somatic mutations in cancerous DNA and cfDNA are related to cervical lymph node metastasis. The cfDNA concentration is related to primary tumor size, lymph node metastasis, and OSCC stage. CONCLUSIONS: Our results show that the detection of TP53 target somatic mutations in OSCC patients by using ddPCR is technically feasible. Low levels of cfDNA may produce different results between cancerous tissue and cfDNA analyses. Future research on cfDNA may quantify diagnostic biomarkers in the surveillance of OSCC patients.

Precise Identification of Recurrent Somatic Mutations in Oral Cancer Through Whole-Exome Sequencing Using Multiple Mutation Calling Pipelines.

Understanding the genomic alterations in oral carcinogenesis remains crucial for the appropriate diagnosis and treatment of oral squamous cell carcinoma (OSCC). To unveil the mutational spectrum, in this study, we conducted whole-exome sequencing (WES), using six mutation calling pipelines and multiple filtering criteria applied to 50 paired OSCC samples. The tumor mutation burden extracted from the data set of somatic variations was significantly associated with age, tumor staging, and survival. Several genes (MUC16, MUC19, KMT2D, TTN, HERC2) with a high frequency of false positive mutations were identified. Moreover, known (TP53, FAT1, EPHA2, NOTCH1, CASP8, and PIK3CA) and novel (HYDIN, ALPK3, ASXL1, USP9X, SKOR2, CPLANE1, STARD9, and NSD2) genes have been found to be significantly and frequently mutated in OSCC. Further analysis of gene alteration status with clinical parameters revealed that canonical pathways, including clathrin-mediated endocytotic signaling, NFκB signaling, PEDF signaling, and calcium signaling were associated with OSCC prognosis. Defining a catalog of targetable genomic alterations showed that 58% of the tumors carried at least one aberrant event that may potentially be targeted by approved therapeutic agents. We found molecular OSCC subgroups which were correlated with etiology and prognosis while defining the landscape of major altered events in the coding regions of OSCC genomes. These findings provide information that will be helpful in the design of clinical trials on targeted therapies and in the stratification of patients with OSCC according to therapeutic efficacy.

Acute spontaneous thoracic epidural hematoma associated with intraspinal lymphangioma: A case report.

BACKGROUND: Spontaneous spinal epidural hematoma is a rare neurosurgical emergency. CASE SUMMARY: A 53-year-old healthy woman suffered from complete paraplegia in both legs and loss of all sensation below the xiphoid process. She was diagnosed as acute spontaneous thoracic epidural hematoma caused by an intraspinal lymphangioma. The primary lab survey showed all within normal limits. Presence of a posteriorly epidural space-occupying lesion at the T4-T8 level of the spinal canal was confirmed on magnetic resonance imaging. A decompressive laminectomy was performed from the T4 to T7 levels at the sixth hour following abrupt onset of complete paraplegia. The lesion was confirmed as lymphangioma. This patient recovered well within one month. CONCLUSION: This study reports a case of acute spontaneous thoracic epidural hematoma caused by an intraspinal lymphangioma with well recovery after surgical intervention.

Occupational radiation exposure of orthopedists at different locations during pedicle screw insertion: An anthropomorphic phantom study.

BACKGROUND: The purpose of this work was to evaluate orthopedic surgeons' exposure to occupational radiation doses from scattering using a mobile flat panel C-arm X-ray machine at different standing positions during an intraoperative pedicle screw implantation. OBJECTIVE: Evaluate the radiation dose received by medical staff, by applying flat X-ray machine in surgical room during an intraoperative pedicle screw implantation. METHODS: A mobile flat-panel C-arm X-ray machine at a dedicated orthopedic operating room was used to image an anthropomorphic female phantom which was set in a prone position on the operating table. The X-ray was projected horizontally, and 1 minute continuous fluoroscopy was used for lumbar spine and thoracolumbar spine during pedicle screw implantation. Scattering radiation doses to orthopedic surgeons were measured at different standing positions and body heights (50, 100, 150 cm above the ground) with and without limited collimations. RESULTS: The dose area product (DAP) in this experiment is normalized as 343 µGy⋅m2. In the four areas, the lowest scattered radiation measured by DF is 11.2 vs. 0.7 µSv, outside and inside the lead suit, respectively, with or without restricted field, 150 cm above the ground, and the lowest scattered radiation dose inside the lead suit. It is 1.3 vs. 0.5 µSv. Comparing the highest dose of the TF at with the lowest dose of the DF, the average result is 73.7 vs. 11.1 µSv, P< 0.05. CONCLUSIONS: Using a mobile flat-panel C-arm X-ray machine during a pedicle screw implantation, the minimum scattering radiation to surgeons was found to be at the terminal DF area based on the analysis of the scattering doses orthopedic surgeons were exposed to.

Overexpression of Platelet-Derived Growth Factor and Its Receptor Are Correlated with Oral Tumorigenesis and Poor Prognosis in Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is a cancerous disease with poor prognosis. According to the statistics, the 5-year survival rate has not improved significantly over the past 20 years. The platelet-derived growth factor (PDGF) and its signaling pathway is a key regulator of angiogenesis and tumorigenesis. High level of PDGF and its receptor (PDGFR) have been reported in several types of malignancies. In this study, we investigated the relationship of the molecular expression levels of PDGF and PDGFR with clinicopathological parameters in OSCC. To this end, we measured the mRNA and protein levels of PDGF and PDGFR by real-time quantitative PCR (qRT-PCR), immunohistochemistry, and enzyme-linked immunosorbent assay (ELISA), respectively. We found positive correlations of the mRNA levels of PDGFA, PDGFB, and PDGFRB with lymph node metastasis and poor overall survival (OS). High expression of PDGF, PDGFRA, and PDGFRB were remarkably associated with lymph node metastasis and poor OS, as determined by immunohistochemistry. Preoperative serum levels of PDGF-AA and PDGF-BB had a positive correlation with preoperative platelet count. Elevated serum levels of PDGF-AA. PDGF-BB, and platelet count correlated with lymph node metastasis and an unfavorable outcome. In multivariate Cox regression analysis, PDGFA mRNA, PDGFB mRNA, PDGFRB mRNA, PDGF immunoexpression, PDGFRB immunoexpression, serum PDGF-AA, serum PDGF-BB, and platelet count emerged as significant independent prognostic factors for OS. In vitro, we found that elevated PDGF promotes colony formation, migration, and invasiveness of SAS and OECM-1 cancer cell lines. Our results suggest that the expression level of serum PDGF has the potential to become a useful diagnostic marker for the prognosis of OSCC. In addition, PDGFR should be considered as a potential therapeutic target for OSCC. Furthermore, research should be undertaken to elucidate the role of PDGF and PDGFR regarding the behavior of tumor cells in OSCC.

SMAD4 Somatic Mutations in Head and Neck Carcinoma Are Associated With Tumor Progression.

As the incidence and the mortality rate of head and neck squamous cell carcinoma (HNSCC) is increasing worldwide, gaining knowledge about the genomic changes which happen in the carcinogenesis of HNSCC is essential for the diagnosis and therapy of the disease. SMAD4 (DPC4) is a tumor suppressor gene. It is located at chromosome 18q21.1 and a member of the SMAD family. Which mediates the TGF-ß signaling pathway, thereby controlling the growth of epithelial cells. In the study presented here, we analyzed tumor samples by multiplex PCR-based next-generation sequencing (NGS) and found deleterious mutations of SMAD4 in 4.1% of the tumors. Knock-down experiments of endogenous and exogenous SMAD4 expression demonstrated that SMAD4 is involved in the migration and invasion of HNSCC cells. Functional analysis of a missense mutation in the MH1 domain of SMAD4 may be responsible for the loss of function in suppressing tumor progression. Missense SMAD4 mutations, therefore, could be useful prognostic determinants for patients affected by HNSCCs. This report is the first study where NGS analysis based on multiplex-PCR is used to demonstrate the imminent occurrence of missense SMAD4 mutations in HNSCC cells. The gene analysis that we performed may support the identification of SMAD4 mutations as a diagnostic marker or even as a potential therapeutic target in head and neck cancer. Moreover, the analytic strategy proposed for the detection of mutations in the SMAD4 gene may be validated as a platform to assist mutation screening.

Establishing of mouse oral carcinoma cell lines derived from transgenic mice and their use as syngeneic tumorigenesis models.

BACKGROUND: The survival of OSCC patient needs to be further improved. miR-211 is oncogenic in OSCC and its upregulation is associated with tumor progression and poor patient survival. K14-EGFP-miR-211 transgenic mice also exhibit augmented potential for OSCC induction. METHODS: Four murine OSCC cell lines, designated MOC-L1 to MOC-L4, are established from tongue tumors induced by 4-nitroquinoline 1-oxide using the K14-EGFP-miR-211 transgenic mouse model. The genetic disruption, in vitro oncogenicity, and the eligibilities of tumorigenesis and metastasis of the cell lines are analyzed. RESULTS: All cell lines show green fluorescence and express a range of epithelial markers. The MOC-L1, MOC-L2 and MOC-L3 cells carry missense mutations in the DNA binding domain of the p53 gene. MOC-L1 exhibits a high level of epithelial-mesenchymal transition and has the aggressive characteristics associated with this. MOC-L1 and MOC-L2 are clonogenic in vitro as well as being tumorigenic when implanted into the dermis or tongue of syngeneic recipients. Nonetheless, only MOC-L1 exhibits immense potential for local regional and distal metastasis. Since the expression of miR-196b in MOC-L1 xenografts is drastically decreased on cisplatin treatment, it would seem that targeting of miR-196b might facilitate tumor abrogation. CONCLUSIONS: As cell lines established in this study originated from the C57BL/6 mouse, the strain most suitable for transgenic engineering, exploring the interplay of these OSCC cells with other genetically modified cells in immune-competent mice would provide important insights into OSCC pathogenesis.

Increased Plasma Circulating Cell-Free DNA Could Be a Potential Marker for Oral Cancer.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is a disease that affects patients worldwide. DNA of dead cells is released into the blood stream and may be isolated from plasma or serum samples. This DNA is termed cell-free DNA (cfDNA). cfDNA is increased in several types of malignancies. We investigated if there was a correlation between cfDNA levels and the progression of OSCC. METHODS: Using quantitative spectrometry, we measured plasma cfDNA in 121 patients with OSCC and 50 matched controls. Mann Whitney and Wilcoxon tests were used to compare differences among various clinical variants. Receiver operating characteristic (ROC) analysis was used to obtain levels suitable for the separation of the clinical subsets. Kaplan-Meier analysis was used to assess correlation with survival. RESULTS: Plasma cfDNA was significantly elevated in patients with OSCC relative to controls. Plasma cfDNA levels correlated with larger tumor size, cervical lymph node metastasis and late stage. Higher plasma cfDNA levels were associated with a poor prognosis of OSCC, which is a new finding. CONCLUSION: Plasma cfDNA could serve as a novel and easily accessible biomarker in OSCC, providing diagnostic and prognostic value.

FAT1 somatic mutations in head and neck carcinoma are associated with tumor progression and survival.

In recent years, the incidence and mortality rates of head and neck squamous cell carcinoma (HNSCC) have increased worldwide. Therefore, understanding genomic alterations in HNSCC carcinogenesis is crucial for appropriate diagnosis and therapy. Protocadherin FAT1, which encodes 4588 amino acid residues, regulates complex mechanisms to promote oncogenesis or suppression of malignancies. Multiplex PCR-based next-generation sequencing (NGS) revealed FAT1 somatic mutations. The clinicopathologic implications of FAT1 in HNSCC were investigated using expression assays, and the functional role of FAT1 in HNSCC pathogenesis was determined using ectopic expression and knockdown experiments. Approximately 29% patients with HNSCC harbored damaging FAT1 mutations. InVEx algorithm identified FAT1 as a significant functional mutation burden. Each type of mutation (missense, nonsense and frameshift) accounted for nearly one-third of deleterious mutations. FAT1 mutations correlated with lower FAT1 expression in tumors. The knockdown of the endogenous expression of FAT1 and exogenous expression of crucial FAT1 domains unequivocally indicated that FAT1 suppressed the migration and invasion capability of HNSCC cells. Functional analysis suggested that nonsense mutations in FAT1 result in the loss of the suppression of tumor progression. FAT1 mutations and downregulation defined nodal involvement, lymphovascular permeation and tumor recurrence. In addition, FAT1 mutations and downregulation are independent predictors of poor disease-free survival in patients with HNSCC. This study is the first to perform multiplex PCR-based NGS to indicate marked non-synonymous FAT1 mutations in HNSCC, which are prognostic indicators. The gene analysis strategy proposed for detecting FAT1 mutations may be a valid method for mutation screening.

Personal Exposure Prescription Method Reduces Dose in Radiography.

PURPOSE: To evaluate the effectiveness of an automatic, personalized exposure prescription method designed to reduce radiation dose during radiography examinations. METHODS: Using standard imaging parameters of average-sized patients, the authors measured individual body-part thicknesses or imaging regions of 116 patients (69 men, 47 women) and calculated each patient's exposure amount according to the thickness of the part or region. The data were used to develop each patient's personalized exposure prescription. Using the personalized exposure prescriptions, authors acquired chest images of the patients on a Carestream DRX-Revolution mobile digital radiography system. RESULTS: All images acquired using the personalized exposure prescription method were satisfactory for diagnosis; exposure indexes were above 1300, a figure deemed acceptable for diagnosis by the manufacturer. The personalized exposure method reduced the amount of radiation each patient received. DISCUSSION: Variation of tube voltage alone can control patients' exposure levels; however, using the personalized exposure prescription method eliminates the need to use automatic exposure controls. CONCLUSION: The personalized exposure prescription method is an effective tool for reducing radiation to patients during radiography as well as for eliminating dose creep.

Effectiveness of a Tailored Anterior Saturation Band in the Improvement of the Image Quality of Pelvic Magnetic Resonance for Assessing Rectal Cancer.

PURPOSE: We sought to test the effectiveness of the application of a tailored anterior saturation band (ASB) to improve the image quality of pelvic magnetic resonance imaging (MRI) for assessing rectal cancer. METHODS: A total of 165 patients with MRI assessment of rectal cancer between 2013 and 2015 were included. The image quality scores (4-point scale: 1, nondiagnostic through 4, excellent) of MRI without and with tailored ASBs were compared. Sensitivity, specificity, positive and negative predictive values, and accuracy of pelvic MRIs with and without a tailored ASB for T-staging in 65 patients with direct surgery and 67 patients with chemoradiotherapy before surgery were evaluated. RESULTS: Two independent raters exhibited moderate-to-excellent interobserver agreements (κ = 0.529-0.879) in the grading of MRI image quality. Overall, the quality scores of sagittal and axial T2-weighted images with tailored ASBs were significantly improved compared with MRIs without ASBs (3.5 ± 0.3 vs. 2.7 ± 0.8 [mean ± SD]; P < .001, and 3.6 ± 0.3 vs. 2.8 ± 0.8; P < .001, respectively). The application of tailored ASBs in MRIs improved the averaged accuracies for staging of ≤ T2, T3, and T4 tumors from 87.7%, 78.5%, and 90.8% to 93.1%, 86.9%, and 97.7%, respectively. In post-chemoradiotherapy MRI follow-ups, the use of tailored ASBs also improved the average accuracies for staging of yT0, yT1-2, yT3, and yT4 tumors from 80.6%, 73.1%, 73.9%, and 91.0%, to 85.8%, 82.9%, 85.1%, and 94.0%, respectively. CONCLUSIONS: Application of a tailored ASB in pelvic MRI is effective in substantially reducing motion artifacts, significantly upgrading image quality, and improving accuracies of rectal tumor staging.

Downexpression of Matriptase-2 Correlates With Tumor Progression and Clinical Prognosis in Oral Squamous-Cell Carcinoma.

The aim of this study was to investigate the relationship of matriptase-2 expression with the clinicopathologic characteristics, the histologic grade, and patient survival in oral squamous-cell carcinoma (OSCC). Immunohistochemical analysis of matriptase-2 expression was performed in 102 surgical specimens from patients with OSCC. The immunohistochemical results were further verified by quantitative real-time reverse transcription-polymerase chain reaction. The immunostaining intensity was scored on a scale ranging from 0 (absence of staining) to 3 (intense staining). The distribution score was determined by the percentage of stained cells on a scale ranging from 0 (<5%), 1 (5% to 25%), 2 (25% to 50%), 3 (50% to 75%), to 4 (75% to 100%). The immunoscore of matriptase-2 expression was the product of the above 2 scores and ranged from 0 to 12 for analysis. Faint matriptase-2 immunostaining was observed in the non-neoplastic oral mucosal epithelia. The matriptase-2 immunoscore was significantly higher in well-differentiated OSCCs than in poorly differentiated tumors (P=0.001). Moreover, a reduced matriptase-2 immunoscore was inversely correlated with the tumor size (P=0.017), a positive nodal stage (P=0.008), distant metastasis (P=0.032), and a late clinical stage (P=0.001). A lower immunoscore of matriptase-2 expression revealed a significant association with poor survival (P=0.003). Our results demonstrate that the inverse expression of matriptase-2 correlates with tumor progression and an advanced TNM stage, and has a poor prognosis in patients with OSCC. These findings suggest that the expression of matriptase-2 may be both a prognostic marker and a potential therapeutic target for this cancer.

Impact of apurinic/apyrimidinic endonuclease 1/redox factor-1 on treatment response and survival in oral squamous cell carcinoma.

BACKGROUND: Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a multifunctional protein involved in DNA repair and redox signaling. The purpose of this study was to investigate the relationship between APE1/Ref-1 expression and clinicopathological features, survival, and treatment response in patients with oral squamous cell carcinoma (OSCC) and cell lines. METHODS: APE1/Ref-1 expression in OSCC was evaluated by immunohistochemistry, and its relationship to patient outcomes and treatment response was assessed statistically. The effects of stable short hairpin (sh)RNA-mediated knockdown of APE1/Ref-1 on cell survival, migration, and chemoradiation sensitivity were determined in OSCC cell lines. RESULTS: APE1/Ref-1 immunostaining was correlated with positive lymph node status, and higher APE1/Ref-1 expression was significantly associated with poor prognosis and reduced treatment response. Consistent with the clinical studies, APE1/Ref-1 expression in OSCC cell lines was implicated in the regulation of migration and cisplatin-induced apoptosis. CONCLUSION: Elevated APE1/Ref-1 expression may be used to predict poor survival and may confer chemoresistance in OSCC.

The hMLH1 -93G>A promoter polymorphism is associates with outcomes in oral squamous cell carcinoma patients.

BACKGROUND: The aim of this study was to investigate the impact of hMLH1 polymorphisms on treatment outcomes in patients with oral squamous cell carcinoma (OSCC). METHODS: Genotypings were performed by direct DNA sequencing in peripheral blood leukocytes from 185 male OSCC patients. Patients received primary surgery with or without adjuvant radiotherapy. Two hMLH1 tag single nucleotide polymorphisms (SNPs)-rs1800734 (-93G>A in the promoter) and rs1540354 (in the third intron)-were chosen from the HapMap project. Overall survival (OS) and disease-free survival (DFS) were compared between different genotypes. RESULTS: The hMLH1 rs1800734 and rs1540354 polymorphisms were in weak linkage disequilibrium (r (2) = 0.456). OSCC patients with the rs1800734 AA genotype had a significantly poor prognosis in both OS and DFS. This SNP can also predict the outcomes of OSCC patients with postoperative adjuvant radiotherapy, especially in advanced stage; however, no significant differences in patient outcomes were found for the hMLH1 rs1540354 genotypes. CONCLUSIONS: Our results demonstrate that the hMLH1 -93G>A SNP is found to be associated with patient outcomes in OSCC. This SNP can also predict their treatment outcome of radiotherapy.

Expression of matriptase correlates with tumour progression and clinical prognosis in oral squamous cell carcinoma.

AIMS: To investigate the relationship of matriptase expression in oral squamous cell carcinoma (OSCC) to clinicopathological characteristics, patient survival and cell-invasive properties. METHODS AND RESULTS: Matriptase expression in OSCC was evaluated by immunohistochemical staining, and its relationship to clinicopathological features and outcomes was assessed statistically. The shRNA-mediated stable knockdown of matriptase in OSCC cells was used to analyse cell proliferation, migration and invasion in vitro. Matriptase immunostaining score was correlated with histopathological grade, clinical stage, positive lymph node and distant metastasis, and higher matriptase immunostaining score was associated significantly with poor prognosis. Elevated matriptase expression in oral cancer cell lines was a significant promoter of oral cancer cell migration and invasion. CONCLUSIONS: Matriptase expression correlates with tumour progression and invasive capability in OSCC and may be an adverse prognostic marker for this cancer.