Metabolomic Analysis Reveals that SPHK1 Promotes Oral Squamous Cell Carcinoma Progression through NF-κB Activation.

BACKGROUND: Metabolic disorders are significant in the occurrence and development of malignant tumors. Changes of specific metabolites and metabolic pathways are molecular therapeutic targets. This study aims to determine the metabolic differences between oral squamous cell carcinoma (OSCC) tissues and paired adjacent noncancerous tissues (ANT) through liquid chromatography-mass spectrometry (LC-MS). SPHK1 is a key enzyme in sphingolipid metabolism. This study also investigates the potential role of SPHK1 in OSCC. MATERIALS AND METHODS: This study used LC-MS to analyze metabolic differences between OSCC tissues and paired ANT. Principal component analysis (PCA) and partial least-squares discriminant analysis (PLS-DA) were applied to explain the significance of phospholipid metabolism pathways in the occurrence and development of OSCC. Through further experiments, we confirmed the oncogenic phenotypes of SPHK1 in vitro and in vivo, including proliferation, migration, and invasion. RESULTS: The sphingolipid metabolic pathway was significantly activated in OSCC, and the key enzyme SPHK1 was significantly upregulated in oral cancer tissues, predicting poor OSCC prognosis. In this study, SPHK1 overexpression was associated with high-grade malignancy and poor OSCC prognosis. SPHK1 targeted NF-κB by facilitating p65 expression to regulate OSCC tumor progression and promote metastasis. CONCLUSIONS: This study identified metabolic differences between OSCC and paired ANT, explored the carcinogenic role of overexpressed SPHK1, and revealed the association of SPHK1 with poor OSCC prognosis. SPHK1 targets NF-κB signaling by facilitating p65 expression to regulate tumor progression and promote tumor metastasis, providing potential therapeutic targets for diagnosing and treating oral tumors.

Anatomical study and clinical application of medial sural artery perforator flap for oral cavity reconstruction.

The present study aims to provide anatomical evidence for clinical application of the medial sural artery perforator (MSAP) flap. The current study investigated the vascular anatomy of the flap, evaluated the postoperative appearance and function of the donor and recipient sites, and investigate the clinical value in reconstruction of oral cavity. Six lower limbs of Chinese adult cadavers were microsurgically dissected. The locations and courses of the medial sural artery perforators were identified and recorded, which provided an anatomical basis for clinical application. Then, 16 clinical cases employing this flap were evaluated, ranging from 3×4cm to 6×8cm, and were employed for defects in the oral cavity region. Sixteen clinical cases with intraoral soft tissue defects, which included four clinical cases with inner cheek defects, were successfully followed up for 10-47 months (24 months on average). The donor site function, contour of recipient site and oral function recovery were evaluated as acceptable or better in cases with intraoral soft tissue defect, which were further verifying the value of clinical application of MSAP in repairing oral cavity defects. Moreover, two typical clinical cases were described in detail. To conclude, the MSAP flap is a favorable choice for small- to medium-size defects based on minor donor site morbidity, satisfactory oral function recovery, perforator stability and adaptation of the pedicle for anastomosis in the oral cavity region.

Knockdown of MSI1 inhibits the proliferation of human oral squamous cell carcinoma by inactivating STAT3 signaling.

Musashi RNA­binding protein 1 (MSI1) is highly expressed in several types of cancer; however, its role in oral squamous cell carcinoma (OSCC) remains unknown. The purpose of this study was to investigate the probable mechanism underlying the involvement of MSI1 in OSCC. The results demonstrated that MSI1 was upregulated in OSCC tissues, but not in adjacent healthy tissues. MSI1 silencing resulted in decreased cell proliferative, invasive and migrative capacity. In addition, MSI1 silencing led to cell cycle arrest at the S phase, downregulation of c­Myc and cyclin D1, and upregulation of p21 and p27 levels. Additional studies demonstrated that MSI1 suppression inhibited the activation of signal transducer and activator of transcription 3 (STAT3) signaling. Accordingly, the findings of the present study suggested that MSI1 silencing can suppress OSCC cell proliferation and progression, in part by inhibiting the activation of the c­Myc/STAT3 pathway.

Inactivation of RUNX3 protein expression in tongue squamous cell carcinoma and its association with clinicopathological characteristics.

The function of runt­related transcription factor 3 (RUNX3) in oral cancer remains controversial. The present study aimed to determine the status of RUNX3 protein expression and its association with clinicopathological characteristics in tongue squamous cell carcinomas (SCC). The present study used three pairs of tongue SCC and non­cancerous tissues to assess RUNX3 protein expression by western blot analysis, and two tongue SCC cell lines to determine RUNX3 protein localization by immunofluorescence and immunocytochemistry. Tissue microarray immunohistochemistry was performed to detect the clinical relevance of RUNX3 in 79 patients with tongue SCC. The results demonstrated that RUNX3 protein expression was reduced in tongue SCC tissues compared with in paired non­cancerous tissues. Similarly, the expression of RUNX3 was low in SCC25 and Cal27 cells, and was predominantly localized to the cytoplasm. In the 79 patients with tongue SCC, RUNX3 protein expression was presented in different manners in carcinoma nests and tumor stroma. RUNX3 in carcinoma nests (nRUNX3) exhibited nuclear positive staining in 24/79 samples, cytoplasmic mislocalization in 41/79 samples and was undetectable in 14/79 samples. RUNX3 in stroma (sRUNX3) exhibited nuclear positive staining in 40/79 samples and nuclear negative staining in 39/79 samples. Negative nRUNX3 expression was significantly associated with lymph node metastasis (P=0.014), clinical stage (P=0.027), and overall and disease­free survival (P=0.008 and P=0.007, respectively). In addition, negative sRUNX3 expression was associated with lymph node metastasis (P=0.003) and clinical stage (P=0.003); however, not with overall survival. The findings of the present study preliminarily suggested that cytoplasmic mislocalization of RUNX3 protein may be a common event in tongue SCC, and that sRUNX3 protein expression may be a potential prognostic biomarker.

RUNX3 plays a tumor suppressor role by inhibiting cell migration, invasion and angiogenesis in oral squamous cell carcinoma.

Although aberrant expression of Runt-related transcription factor 3 (RUNX3) contributes to tumor progression and metastasis in a number of carcinomas, the status of RUNX3 and its correlation with prognosis in oral squamous cell carcinomas (OSCC) are still controversial. The aim of present study was to investigate the function of RUNX3 in OSCC and the underlying molecular mechanisms. Tissue microarray (TMA) consisting of 232 OSCC specimens was used to detect the expression of RUNX3 by immunohistochemistry method. The effects of RUNX3 restoration on OSCC cell migration and invasion were determined by wound-healing assay, migration and Matrigel cell invasion assays. The antiangiogenic role of RUNX3 was analyzed by testing proliferation and tube formation of human umbilical vascular endothelial cells (HUVECs) cultured with conditioned medium from RUNX3 transfected OSCC cell lines. The activities of MMP-9 and VEGF in RUNX3 transfected OSCC cell lines were examined by western blot and Elisa methods. RUNX3 expression was reduced in OSCC specimens and significantly associated with tumor size (P=0.002), lymph node statue (P=0.0036) and clinical stage (P=0.0001). Negative expression of RUNX3 correlated with worse 5-year overall and disease-specific survival rates (P=0.0348 and P=0.0301, respectively). Furthermore, we found that RUNX3 restoration suppressed cell migration and invasion through downregulating MMP-9 expression and secretion, and exerted antiangiogenic capability by inhibiting VEGF activity in HN6 and Cal27 cells. These findings suggested that RUNX3 played a tumor suppressor role in OSCC by inhibiting cell migration, invasion and angiogenesis, supporting that it could be a potential therapeutic target for OSCC.

Therapeutic effects of tyroservatide on metastasis of lung cancer and its mechanism affecting integrin-focal adhesion kinase signal transduction.

Tyroservatide (YSV) can inhibit the growth and metastasis of mouse lung cancer significantly. This study investigated the therapeutic effects of tripeptide YSV on metastasis of human lung cancer cells and explored its possible mechanism that affects integrin-focal adhesion kinase (FAK) signal transduction in tumor cells. YSV significantly inhibited the adhesion and the invasion of highly metastatic human lung cancer cell lines 95D, A549, and NCI-H1299. In addition, YSV significantly inhibited phosphorylation of FAK Tyr397 and FAK Tyr576/577 in the 95D, A549, and NCI-H1299 human lung cancer cells in vitro. And the mRNA level and protein expression of FAK in these human lung cancer cells decreased at the same time. YSV also significantly inhibited mRNA and protein levels of integrin ß1 and integrin ß3 in the 95D, A549, and NCI-H1299 human lung cancer cells. Our research showed that YSV inhibited adhesion and invasion of human lung cancer cells and exhibited therapeutic effects on metastasis of lung cancer.

Prognostic role of microRNA-145 in various human malignant neoplasms: a meta-analysis of 18 related studies.

BACKGROUND: Recent studies show that microRNA-145 (miR-145) might be an attractive tumor biomarker of considerable prognostic value. To clarify the preliminary predictive value of miR-145 for prognosis in various malignant neoplasms, we conducted a meta-analysis of 18 relevant studies. METHODS: Eligible studies were identified by searching the online databases PubMed, EMBASE, and Web of Science up to March 2014. Pooled hazard ratios (HRs) with 95% confidence intervals (CIs) for patient survival and disease progress were calculated to investigate the association with miR-145 expression. RESULTS: In total, 18 eligible studies were included in this meta-analysis. Our results showed that upregulated miR-145 significantly predicted a favorable overall survival (OS) (HR = 0.47, 95% CI 0.31 to 0.72), but failed to show a significant relation with disease prognosis. In stratified analyses, high miR-145 expression predicted favorable OS in both Whites and Asians but the intensity of the association in Whites (HR = 0.67, 95% CI 0.47 to 0.95) was not as strong as in Asians (HR = 0.35, 95% CI 0.19 to 0.64). High miR-145 expression also predicted better progression-free survival (PFS) in Asians (HR = 0.43, 95% CI 0.21 to 0.89), but not in Whites. In addition, a significantly favorable OS associated with upregulated miR-145 expression was observed in both squamous cell (SCC) (HR = 0.34, 95% CI 0.13 to 0.93) and glioblastoma (HR = 0.72, 95% CI 0.52 to 0.99). CONCLUSIONS: Our findings indicate that high miR-145 expression is better at predicting patient survival rather than disease progression for malignant tumors, especially for SCC and glioblastoma in Asians. Considering the insufficient evidence, further investigations and more studies are needed.

The combination of SMAD4 expression and histological grade of dysplasia is a better predictor for the malignant transformation of oral leukoplakia.

Oral leukoplakia (OL) is the most common premalignancy in the oral cavity and can progress to oral squamous cell carcinoma (OSCC). SMAD4 is a tumor suppressor implicated in multiple cancer types including OSCC. To assess the role of SMAD4 in oral leukoplakia malignant transformation, the authors investigated SMAD4 expression patterns in OL and OSCC using a highly specific antibody and correlated the patterns with the risk of malignant transformation oral leukoplakia. Immunohistochemistry and a quantitative imaging system were used to measure SMAD4 expression in OL from 88 OL patients, including 22 who later went through malignant transformation, and their OSCC counterpart. Forty-three (48.9%) of the 88 OL patients had strong SMAD4 expression. SMAD4 expression had no significant correlation with patients' clinicopathological parameters. Interestingly, 17 (39.5%) of the 43 OL lesions with strong SMAD4 expression went through malignant transformation whereas only 5 (11.1%) of the 45 OL lesions with weak SMAD4 expression did so (p = 0.002). The SMAD4 expression in OL was much higher than that in their OSCC counterpart. Kaplan-Meier analysis revealed that the combination of SMAD4 expression and histological grade of dysplasia (p = 0.007) is a better predictor for the malignant transformation of oral leukoplakia. In the multivariate analysis, both SMAD4 expression and grade of dysplasia were identified as independent factors for OL malignant transformation risk (p = 0.013 and 0.021, respectively). It was concluded that high SMAD4 expression may be indicative of an early carcinogenic process in OL and serve as an independent biomarker in assessing malignant transformation risk in patients with OL, and the combination of SMAD4 expression and histological grade of dysplasia is a better predictor for the malignant transformation of oral leukoplakia.

[Effect of jaw forward distance on forced inspiratory airflow in patients with obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To study the effect of different jaw forward distance on forced inspiratory airflow(FIF) in non-apnea subjects and patients with obstructive sleep apnea hypopnea syndrome (OSAHS) and to evaluate the effective jaw forward distance for the treatment of OSAHS with the oral appliance. METHODS: FIF was measured in 18 non-apnea subjects and 18 OSAHS patients at supine and lateral body positions with different jaw forward distances (the percentages of maximum jaw forward distance): 0%, 25%, 50% and 75%. FIF were converted to percentage values (FIF%, x(-) ± s) followed by averaged. Then the results were analyzed by one-way analysis of variance and paired t-test with α = 0.05. RESULTS: For non-apnea subjects, there was no significant difference in the FIF values between different jaw forward distances as well as different body positions. For OSAHS patients, the mean FIF% at supine and lateral body positions were 107.1% ± 29.0% and 112.0% ± 33.1% at jaw forward 50%, and were 106.4% ± 20.7% and 116.8% ± 36.4% at jaw forward 75%, respectively, which were significantly higher than those (84.0% ± 18.3% and 98.3% ± 24.0%) at jaw forward 0% or those (92.7% ± 21.8% and 103.7% ± 22.6%) at jaw forward 25%, respectively. But there was no statistical difference in FIF between the two groups of jaw forward 50% and jaw forward 75% and no statistical difference in FIF between supine and lateral body positions in the same forward position. CONCLUSION: Jaw forward 50% is a effective jaw forward distance by oral appliance for the treatment of OSAHS and can improve the airway ventilation in OSAHS patients.

Bonding property of two resin-reinforced glass-ionomer cements to zirconia ceramic.

OBJECTIVES: To investigate whether a stable bond could be obtained between resin-reinforced glass-ionomer cement and zirconia ceramic. METHOD AND MATERIALS: Sixty disk specimens of a dental ceramic (Cercon zirconia ceramic, Dentsply) were made and treated by airborne-particle abrasion. They were divided into three groups and bonded to two resin-reinforced glass-ionomer cements (RelyX Luting [3M ESPE] and Fuji Plus [GC]) and one resin cement (Panavia F, Kuraray) as a control group. All bonded specimens of each group (n = 20) were stored in distilled water (37 degrees C) for 24 hours, and half were additionally aged by thermocycling (20,000 times). Shear bond strength test was performed to measure the bond strength. Statistical analyses were performed using one-way ANOVA and paired t test with a = .05. The interfacial morphology of debonded specimens was observed by using a scanning electron microscope, and the mode of bonding fracture was evaluated. RESULTS: The initial shear bond strength (in 24 hours) of the two resin-reinforced glass-ionomer cements to zirconia ceramic was 17.33 +/- 3.53 MPa and 16.68 +/- 2.76 MPa, and it dropped significantly to 7.62 +/- 2.17 MPa and 4.65 +/- 2.02 MPa after thermocycling. In the control group, the initial shear bond strength was 26.25 +/- 5.61 MPa, and there was no obvious decrease after thermocycling. The bonding failure of resin-reinforced glass-ionomer cements was mostly adhesive failure between cement and ceramic. CONCLUSION: Resin-reinforced glass-ionomer cement could not offer a stable bond to abraded zirconia ceramic after thermocycling, and there was no durable chemical or mechanical bond between resin-reinforced glass-ionomer cement and zirconia ceramic.

Radial forearm free flap for reconstruction of a large defect after radical ablation of carcinoma of the tongue and floor of the mouth: some new modifications.

BACKGROUND: A modified radial forearm free flap was designed to rehabilitate function and to reduce the complications at both donor and recipient sites. METHODS: Between 2003 and 2007, 15 patients with infiltrating squamous cell carcinoma (T(3)-T(4)) of the tongue and/or floor of the mouth underwent hemiglossectomy and resection of the floor of the mouth with microvascular reconstruction using a modified radial forearm flap. The mean size of the forearm flap was 7.5 x 14 cm, and the de-epithelialized area was 7 x 6 cm, requiring no skin graft from the abdomen. Speech intelligibility tests were administered to test postoperative speech and the functional oral intake scale was applied to assess the postoperative swallowing function, and patients reconstructed with pectoralis major myocutaneous flap were used for comparison. RESULTS: All the flaps were successfully transferred. No obvious complications were found in either the oral-maxillofacial or forearm region. The speech intelligibility was better in the modified flap group (p < 0.01). An acceptable swallowing function was also achieved, although the difference was not significant (p > 0.05). CONCLUSIONS: The modified flap used for reconstructing large defects of the tongue and floor of the mouth might be a valid substitute for pectoralis major myocutaneous flap to improve the outcome in individuals with significant oral carcinoma.

Application of lateral arm free flap in oral and maxillofacial reconstruction following tumor surgery.

OBJECTIVE: To describe the application of lateral arm free flap (LAFF) in reconstruction of defects in the oral and maxillofacial regions following ablative oncological surgery. SUBJECTS AND METHODS: The study included 16 patients (13 male, 3 female, mean age 56, range 35-69 years). Sixteen LAFF were harvested to reconstruct defects caused by the dissection of malignant tumors of the oral and maxillofacial regions. The tumor was squamous cell carcinoma of the tongue (6 cases), floor of the mouth (4), retromolar area (3), inner cheek (2), and lower gingival (1). Flap sizes ranging from 5 x 7 to 6 x 9 cm were harvested using a sterile tourniquet for bloodless technique. The anastomoses were carried out using a magnifier or microscope. All donor defects were closed primarily. RESULTS: Fourteen flaps healed without venous insufficiency. One flap, in a female patient, survived with mild local microcirculatory obstruction but that of another female patient developed necrosis. There was no significant complication at the donor sites. The advantages of this flap include anatomically reliable vascular supply, accessible donor site, and the aesthetic quality of donor tissue is good. Compared with the radial artery, the posterior radial collateral artery is a nonessential vessel of the arm. The disadvantages are the relatively smaller vessel size for anastomosis and thicker subcutaneous tissue. CONCLUSIONS: For the repair of moderate-sized defects of the maxillofacial area, especially in male patients, the LAFF can be recommended.

[Effect of human semaphorin-3F gene transient transfection on the proliferation of tongue carcinoma cells].

OBJECTIVE: To investigate the effect of semaphorin-3F (SEMA-3F) gene transient transfection on the proliferation of Tca8113 tongue carcinoma cells. METHODS: After construction of a full-length SEMA3F expression vector, Tca8113 cells were transient transfected with pEGFP-N1-SEMA3F by Lipofectamine 2000. The expression of SEMA-3F gene was detected by RT-PCR The differences of the expression in the transfected cell groups, empty vector groups and un-transfected cell groups were compared. The survival rates were assayed by methyl thiazolyl tetrazolium (MTT) enzymatic labeling technique. Cell cycle were assayed by flow cytometer (FCM). RESULTS: The gene was transfected into Tca8113 cells. High expression of SEMA-3F was successfully detected in the transfected cell groups after gene transfection. The cell cycle percentage of G1 stage decreased and S stage increased. CONCLUSIONS: SEMA-3F gene transient transfection may inhibit the proliferation of Tca8113 cells.