Erratum: Involvement of miR-619-5p in resistance to cisplatin by regulating ATXN3 in oral squamous cell carcinoma: Erratum.

[This corrects the article DOI: 10.7150/ijbs.54014.].

TNFAIP2 confers cisplatin resistance in head and neck squamous cell carcinoma via KEAP1/NRF2 signaling.

BACKGROUND: Drug resistance limits the treatment effect of cisplatin-based chemotherapy in head and neck squamous cell carcinoma (HNSCC), and the underlying mechanism is not fully understood. The aim of this study was to explore the cause of cisplatin resistance in HNSCC. METHODS: We performed survival and gene set variation analyses based on HNSCC cohorts and identified the critical role of tumor necrosis factor alpha-induced protein 2 (TNFAIP2) in cisplatin-based chemotherapy resistance. Half-maximal inhibitory concentration (IC50) examination, colony formation assays and flow cytometry assays were conducted to examine the role of TNFAIP2 in vitro, while xenograft models in nude mice and 4-nitroquinoline N-oxide (4NQO)-induced HNSCC models in C57BL/6 mice were adopted to verify the effect of TNFAIP2 in vivo. Gene set enrichment analysis (GSEA) and coimmunoprecipitation coupled with mass spectrometry (Co-IP/MS) were performed to determine the mechanism by which TNFAIP2 promotes cisplatin resistance. RESULTS: High expression of TNFAIP2 is associated with a poor prognosis, cisplatin resistance, and low reactive oxygen species (ROS) levels in HNSCC. Specifically, it protects cancer cells from cisplatin-induced apoptosis by inhibiting ROS-mediated c-JUN N-terminal kinase (JNK) phosphorylation. Mechanistically, the DLG motif contained in TNFAIP2 competes with nuclear factor-erythroid 2-related factor 2 (NRF2) by directly binding to the Kelch domain of Kelch-like ECH-associated protein 1 (KEAP1), which prevents NRF2 from undergoing ubiquitin proteasome-mediated degradation. This results in the accumulation of NRF2 and confers cisplatin resistance. Positive correlations between TNFAIP2 protein levels and NRF2 as well as its downstream target genes were validated in HNSCC specimens. Moreover, the small interfering RNA (siRNA) targeting TNFAIP2 significantly enhanced the cisplatin treatment effect in a 4NQO-induced HNSCC mouse model. CONCLUSIONS: Our results reveal the antioxidant and cisplatin resistance-regulating roles of the TNFAIP2/KEAP1/NRF2/JNK axis in HNSCC, suggesting that TNFAIP2 might be a potential target in improving the cisplatin treatment effect, particularly for patients with cisplatin resistance.

Development and validation of a nomogram to predict occult cervical metastasis in early oral squamous cell carcinoma.

Background: Lack of adequate objectivity and universality, available models are still difficult to be applied to clinical practice in predicting occult cervical metastasis of early oral squamous cell carcinoma (OSCC). Taking abnormal metabolic state into consideration, the current model is helpful to distinguish those patients with or without occult cervical metastasis. Methods: This study retrospectively analyzed 330 OSCC patients initially diagnosed cT1-2N0M0 stage and received neck dissection from January 2020 to July 2022. The occult cervical metastasis was identified by pathological examination.. After screening independent risk factors using logistic regression, patients were divided into training and validation cohorts at the ratio of 2:1 randomly, and a novel diagnostic model was constructed. Performances of this model were evaluated by the area under the curve (AUC), calibrating curve, decision curve analysis (DCA) and clinical impact curve (CIC). Results: Of the 330 included patients {age mean [standard deviation (SD)], 61.24 (12.99) years; 202 (61.2%) males}, 49 (14.8%) had occult nodal metastasis. Five variables, including body mass index (BMI) [high odds ratio (OR): 1.132; 95% confidence interval (CI): 1.019-1.258, P=0.021], primary tumor site (tongue & floor of mouth (TF) OR: 3.756; 95% CI: 1.295-10.898, P=0.015), depth of invasion (DOI) (5-10 mm OR: 2.973; 95% CI: 1.266-6.981; P=0.012), pathological differentiation (Poor differentiation OR: 2.65; 95% CI: 1.341-5.239; P=0.005), and diabetes (OR: 3.123; 95% CI: 1.23-7.929; P=0.017) were screened to establish the predictive model. In training cohort (n=220), this model achieved an AUC of 0.814 and had a sensitivity of 78.1% and specificity of 70.2%. Calibration plots showed favorable consistency between the prediction of the model and actual observations (Hosmer-Lemeshow value >0.05). Decision curve analysis (DCA) and clinical impact curve (CIC) showed the model was clinically useful and had better discriminative ability under the threshold probability of 0.5. Above evaluations were verified in the validation cohort (n=110). Compared to previous reported models, the concordance index (C-index), net reclassification index (NRI), and integrated discrimination improvement (IDI) values were superior in both training and validation cohorts (P<0.05). Conclusions: This constructed model might have reference value for clinicians in making neck management decisions of early OSCC patients.

Regulation of Semaphorin3A in the process of cutaneous wound healing.

Semaphorin 3A (Sema3A) has been recognized as a crucial regulator of morphogenesis and homeostasis over a wide range of organ systems. However, its function in cutaneous wound healing is poorly understood. In our study, we demonstrated that Sema3A adenovirus plasmids transfection limited keratinocyte proliferation and decreased migrative capacity as assessed by in vitro wound healing assay. Sema3A transduction inhibited TGF-ß1-mediated keratinocyte migration and EMT process. Besides, we applied mice with K14-Cre-mediated deletion of Sema3A and found that Sema3A depletion postponed wound closure with decreased re-epithelialization and matrix growth. Contrary to the results obtained with full-length Sema3A plasmids transfection, increased keratinocyte migration with recombinant Sema3A proteins resulted in quicker closure of the wounding area after a scratch. Further, exogenously applied recombinant Sema3A worked with EGF to maintain the activation of EGFR by interacting with NRP1 and thereby regulated the internalization of the EGFR-NRP1 complex. Taken together, these results indicated a paradoxical role of autonomous and non-autonomous Sema3A expression during wound healing. Combined administration of recombinant EGF and Sema3A proteins could accelerate the process of wound repair, thus providing promising treatment prospects in the future.

Perineural invasion, lactate dehydrogenase, globulin, and serum sodium predicting occult metastasis in oral cancer.

OBJECTIVE: This study aimed to develop a nomogram to predict the neck occult metastasis in early (T1-T2 cN0) oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: The nomogram was developed in a training cohort of 336 early OSCC patients and was validated in a validation cohort including 88 patients. Independent predictors were calculated by univariate and multivariate logistic regression analyses. RESULTS: In univariate logistical regression analysis, gender, perineural invasion (PNI), blood vessel invasion, mean corpuscular hemoglobin, aspartate aminotransferase, prealbumin, globulin (GLO), lactate dehydrogenase (LDH), serum sodium (NA), and serum chloride were significant associated with neck occult metastasis. Multivariate logistical regression analysis identified PNI (p < .001), LDH (p = .003), GLO (p = .019), and NA (p = .020) as independent predictors of neck occult metastasis. Cut-off values for LDH, GLO, and NA obtained from AUC were 142.5, 26.35, and 139.5, respectively. The nomogram based on PNI and categorical GLO, LDH, and NA exhibited a strong discrimination, with a C-indexes of 0.748 (95%CI = 0.688 to 0.810) in the training cohort and 0.751 (95%CI = 0.639 to 0.863) in the validation cohort. CONCLUSIONS: A nomogram based on PNI, LDH, GLO, and NA for predicting the risk of neck lymph nodes occult metastasis in OSCC could help surgeons with therapy decision-making.

[Application of 3D printing technology under three-dimensional reconstruction in mandibular reconstruction].

PURPOSE: To explore the application value of 3D printing technology under three-dimensional reconstruction in mandibular reconstruction. METHODS: Eighty-four patients with mandibular defect reconstruction were divided into two groups by different operation methods: 3D group(n=42) and control group(n=42). Patients in the control group underwent routine operation, while patients in the experimental(3D) group underwent three-dimensional reconstruction with 3D printing technology. The operation conditions, incidence of complications, recovery of facial features and occlusal relationship were recorded. SPSS 23.0 software package was used for statistical analysis of the data. RESULTS: The operation time of 3D group was significantly shorter than that of the control group, and the amount of bleeding was significantly less than that of the control group(P<0.05). The recovery rate of facial appearance and occlusal relationship in 3D group was significantly higher than in the control group(95.24% vs 78.57%, P<0.05). Compared with the control group, the movement distance of mandibular points in 3D group was significantly smaller before and after operation(P<0.05). The satisfaction scores of chewing function and pronunciation recovery in the two groups were close(P>0.05), but compared with the control group, the satisfaction scores of appearance recovery in the 3D group were significantly higher(P<0.05). CONCLUSIONS: 3D reconstruction under 3D printing technology can reduce intraoperative bleeding, shorten the operation duration, and achieve good shape recovery with high degree of satisfaction.

Involvement of miR-619-5p in resistance to cisplatin by regulating ATXN3 in oral squamous cell carcinoma.

MicroRNAs are major post-transcriptional regulators responsible for the development of human cancers, including OSCC. The specific role of miR-619-5p in OSCC, however, is rarely reported. Cisplatin is one of the mostly applied chemotherapy drugs of OSCC. Nevertheless, drug resistance of cisplatin following the initial chemotherapy largely restricts its clinical benefits, and the mechanism of cisplatin resistance is unclear. This study intends to explore the biological function of miR-619-5p in the development of cisplatin resistance in OSCC cell lines and a xenograft model, as well as the potential molecular mechanism. Our results showed that miR-619-5p was down-regulated in OSCC samples and cisplatin-resistant OSCC cells. Ectopically expressed miR-619-5p inhibited proliferative, migratory and invasive abilities of OSCC cisplatin-resistant cells. The putative target gene ATXN3 was predicted by bioinformatic analysis and confirmed by dual-luciferase reporter assay. Importantly, ATXN3 was responsible for the regulatory effects of miR-619-5p on biological behaviors of cisplatin-resistant OSCC cells. Moreover, miR-619-5p mimics and ATXN3-siRNA significantly enhanced ATXN3 knockdown in both HN6/CDDPR and CAL27/CDDPR cells and inhibited expression of PI3K and AKT. In vivo evidences demonstrated that intratumoral injection of miR-619-5p agomir remarkably slowed down the growth of OSCC in xenograft mice. Collectively, microRNA-619-5p was the vital regulator for regulating cisplatin resistance of OSCC, which may be served as a potential therapeutic target.

Prognostic value of log odds of positive lymph nodes in patients with resectable oral squamous cell carcinoma.

OBJECTIVES: Log odds of positive lymph nodes (LODDS) was reported to be significantly associated with prognosis in several malignant tumors. However, few are the studies on the correlation between LODDS and overall survival (OS) in patients with oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: A retrospective study including 233 patients with OSCC during 2009 to 2013 was conducted. We probed the correlation between clinicopathological factor, LODDS, lymph node ratio (LNR), pN and OS. The potential prognostic factor and the independent factor were calculated using univariate analysis and multivariate analysis respectively. The goodness of fit and the discriminability was analyzed with Somer's D value, Nagelkerke R2 index, and Akaike information criterion (AIC). Kaplan-Meier survival curve of OS was contrasted by log-rank test in LODDS, LNR and pN, respectively. RESULTS: According to the X-tile, the cut-off values are -1.491 and -0.763 for LODDS, 0.024 and 0.133 for LNR. LODDS, LNR and pN were significantly correlated with OS by univariate analysis (P < 0.001). Multivariate analysis demonstrated LODDS, LNR and pN as an independent prognostic factors for OS (P < 0.01). Compared with pN and LNR models, LODDS showed the strongest predictive power. LODDS was superior to LNR and pN in predicting outcomes in patients with no positive lymph nodes and inadequate neck dissection. CONCLUSION: LODDS would be incorporated into future N classification, which may be conducive to discern the prognosis of OSCC and make a decision of adjuvant therapy in clinical practice.

Prognostic value of serum liver enzymes in oral and oropharynx squamous cell carcinomas.

BACKGROUND: Serum liver enzymes, which catalyze relevant catabolic pathways, have been indicated to be diagnostic and prognostic tools for several malignant tumors. The correlation between serum liver enzymes levels and survival in patients with oral and oropharynx squamous cell carcinomas (OSCC) is still absent. Here, we conducted a study focusing on predictive value of serum liver enzymes in terms of prognosis in the patients. METHODS: A retrospective study including 134 OSCC patients from years 2009 to 2014 was performed to investigate the association between levels of pre-treatment serum liver enzymes, various clinical parameters and prognostic outcomes, which are overall survival (OS) and disease-free survival (DFS). Log-rank tests with Kaplan-Meier method were used to detect potential prognostic biomarkers. Multivariate analyses by Cox proportional hazards model were used to identify significant predictors of prognosis. RESULTS: Serum adenosine deaminase (ADA) level was associated with patients' OS and DFS by univariate analyses (P = 0.006 and P = 0.024, respectively). Multivariate analyses showed that higher serum ADA (>17.2 U/L) (P = 0.019) as well as positive lymph node status (P = 0.035) independently predicted worse OS of patients with OSCC. In addition, older age (≥60 years) (P = 0.043) and positive lymph node status (P = 0.027) were independently prognostic parameters for poorer DFS. CONCLUSIONS: Pre-operative serum ADA levels may serve as a reliable independent prognostic predictor for OS in OSCC patients.

Membrane-tethered Notch1 exhibits oncogenic property via activation of EGFR-PI3K-AKT pathway in oral squamous cell carcinoma.

Notch proteins are highly conserved cell surface receptors which play essential roles in cellular differentiation, proliferation, and apoptotic events at all stages of development. Recently, NOTCH1 mutations have been extensively observed in oral squamous cell carcinoma (OSCC) and are hinted to be Notch1-inactivating mutations. However, little is known about the biological effect of these reported mutations in OSCC. To mimic the inactivation of Notch1 due to inappropriate mutations and to determine the potential mechanisms, we utilized wild-type Notch1 vectors (Notch1WT ) or mutant Notch1 vectors (Notch1V1754L ) to transfect into OSCC cell lines. Membrane-tethered Notch1 induced by mutation was analyzed by immunofluorescence staining. γ-Secretase inhibitor PF-03084014 was utilized to determine the phenotype in the absence of endogenous Notch1 activation. Here we demonstrated that membrane-tethered Notch1 inactivated the canonical Notch1 signaling and oncogenic phenotypes were identified by promoting cell proliferation and invasion and by inducing epithelial-to-mesenchymal transition in cells. The γ-secretase inhibitor PF-03084014 also showed distinct oncogenic property after treatment. Importantly, both membrane-tethered Notch1 and PF-03084014 inhibitor activated the epidermal growth factor receptor (EGFR)-phosphoinositide 3-kinase (PI3K)-protein kinase B (AKT) signaling pathway, which has been confirmed as an overwhelming modulator in OSCC. This was the first time that we clearly simulated the mutated Notch1 activities and determined the oncogenic phenotypes of membrane-tethered Notch1. Compared with wild-type Notch1, membrane-tethered Notch1 was strongly associated with activated EGFR-PI3K-AKT signaling pathway.

Mechanical versus Hand-Sewn Venous Anastomoses in Free Flap Reconstruction: A Systematic Review and Meta-Analysis.

BACKGROUND: Venous complications are the primary reason for flap loss in massive defect reconstructions; therefore, the quality and reliability of microvascular anastomoses are significant. The aim of this systematic review was to evaluate venous anastomotic time, the venous complication rate, and the flap failure rate with the mechanical anastomotic coupling device versus the hand-sewn technique in venous anastomoses of microvascular free flap operations. METHODS: Chinese and English databases were searched for eligible articles published between their inception and July of 2017. The pooled relative risk was calculated for dichotomous variables, and the weighted mean difference was calculated for continuous data. Whether to use the fixed effects or random effects model depended on the heterogeneity evaluation among the studies. RESULTS: Twelve studies were selected, including 3788 flaps (mechanical anastomotic coupling device, n = 1667; hand-sewn, n = 2121). Using the mechanical anastomotic coupling device significantly decreased venous anastomotic time (weighted mean difference, -13.50; 95 percent CI, -17.09 to -9.91; p < 0.01) and the incidence of venous complications (relative risk, 0.40; 95 percent CI, 0.25 to 0.65; p < 0.01). There was a significant difference in terms of flap failure between the groups (relative risk, 0.56; 95 percent CI, 0.32 to 0.97; p = 0.04); thus, flap survival improved with the assistance of the mechanical anastomotic coupling device. No publication bias was detected in those analyses. CONCLUSION: This meta-analysis suggests that the mechanical anastomotic coupling device contributes to reduced operative time, decreased probability of surgical reexploration, and mitigation of flap loss.

Serum bilirubin level predicts postoperative overall survival in oral squamous cell carcinoma.

BACKGROUND: Aberrant level of serum bilirubin, marker of hepatobiliary and hematological disorders, was associated with patient prognosis in several human malignancies. In this study, we aim to evaluate the predictive value of serum bilirubin for clinicopathologic characteristics and survival of patients with oral squamous cell carcinoma (OSCC). METHODS: This study retrospectively reviewed 129 patients with OSCC and 129 normal controls matched for age and sex. The association between levels of preoperative direct bilirubin (DBIL), indirect bilirubin (IBIL), total bilirubin (TBIL), and clinical variables was analyzed. A proportional hazards regression model was used to find out the independent predictors of survival. RESULTS: Significantly lower TBIL (P = .009) and IBIL (P < .001) were found in OSCC patients compared with normal controls. DBIL (P = .011) and lymph-node metastasis (P = .031) were found to be independent prognostic factors. Patients with higher DBIL (≥4.0 µmol/L) had longer overall survival than those with lower DBIL (P = .002). Patients with both lymph-node metastasis and lower DBIL showed the shortest overall survival (P = .001). CONCLUSIONS: Lower DBIL was associated with a poorer prognosis and may be regarded as an independent prognostic marker for patients with OSCC.