Targeted Knockdown of Macrophage Migration Inhibitory Factor Enhances UVB Irradiation-Induced Apoptosis Via Increasing ROS Generation in Oral Squamous Cell Carcinoma.

Objectives: We investigated the effects of macrophage migration inhibitory factor (MIF) knockdown or overexpression combined with ultraviolet radiation B (UVB) irradiation on cell proliferation and apoptosis of oral squamous cell carcinoma (OSCC). Methods: MIF expression in OSCC and adjacent tissues was detected by immunohistochemistry. MIF expression in human immortalized oral epithelial cells (HIOEC) and OSCC cells was detected by western blotting. MIF was knocked down or overexpressed in OSCC cell lines (SCC-25 and CAL-27). OSCC cells were set up into control (CON), MIF overexpression/knockdown (oeMIF/shMIF), CON + UVB, and oeMIF + UVB/shMIF + UVB groups based on their exposure to UVB irradiation. Cell line proliferation was studied using a cell counting kit-8 (CCK-8) and colony formation assays. Flow cytometry was applied for determination of apoptosis, cell cycle, reactive oxygen species (ROS) abundance, and mitochondrial membrane potential. Apoptosis-related proteins were assayed by western blotting. Results: The expression of MIF was significantly higher in OSCC tissues and cell lines than in adjacent tissues and HIOEC. MIF knockdown accompanied by UVB irradiation significantly hampered cell viability and proliferation compared to MIF knockdown or UVB irradiation alone. Western blotting and flow cytometry showed that MIF knockdown combined with UVB irradiation not only induced apoptosis via the mitochondrial pathway but also mediated the cell cycle. Flow cytometry showed that ROS and mitochondrial membrane potential depolarization were increased in the combination treatment groups compared with the mono-treatment groups. Additionally, the ROS scavenger N-acetylcysteine significantly attenuated MIF knockdown combined with UVB irradiation-induced apoptosis and reversed MIF knockdown combined with UVB irradiation-induced MAPK activation. Conclusion: MIF knockdown combined with UVB irradiation significantly inhibited the proliferation of OSCC cells. MIF was involved in UVB-induced ROS generation and enhanced UVB irradiation-induced mitochondria-dependent apoptosis of OSCC cells by activating the MAPK pathway. This suggests that MIF-targeted therapy combined with UVB irradiation may be a novel approach for treating OSCC.

Predicting prognosis of nasopharyngeal carcinoma based on deep learning: peritumoral region should be valued.

BACKGROUND: The purpose of this study was to explore whether incorporating the peritumoral region to train deep neural networks could improve the performance of the models for predicting the prognosis of NPC. METHODS: A total of 381 NPC patients who were divided into high- and low-risk groups according to progression-free survival were retrospectively included. Deeplab v3 and U-Net were trained to build segmentation models for the automatic segmentation of the tumor and suspicious lymph nodes. Five datasets were constructed by expanding 5, 10, 20, 40, and 60 pixels outward from the edge of the automatically segmented region. Inception-Resnet-V2, ECA-ResNet50t, EfficientNet-B3, and EfficientNet-B0 were trained with the original, segmented, and the five new constructed datasets to establish the classification models. The receiver operating characteristic curve was used to evaluate the performance of each model. RESULTS: The Dice coefficients of Deeplab v3 and U-Net were 0.741(95%CI:0.722-0.760) and 0.737(95%CI:0.720-0.754), respectively. The average areas under the curve (aAUCs) of deep learning models for classification trained with the original and segmented images and with images expanded by 5, 10, 20, 40, and 60 pixels were 0.717 ± 0.043, 0.739 ± 0.016, 0.760 ± 0.010, 0.768 ± 0.018, 0.802 ± 0.013, 0.782 ± 0.039, and 0.753 ± 0.014, respectively. The models trained with the images expanded by 20 pixels obtained the best performance. CONCLUSIONS: The peritumoral region NPC contains information related to prognosis, and the incorporation of this region could improve the performance of deep learning models for prognosis prediction.

In vitro assessment of the role of DpC in the treatment of head and neck squamous cell carcinoma.

The present study aimed to investigate the antitumor efficacy of di-2-pyridylketone-4-cyclohexyl-4-methyl-3-thiosemicarbazone (DpC) and di-2-pyridylketone-4,4,-dimethyl-3-thiosemicarbazone (Dp44mT) on head and neck squamous cell carcinoma (HNSCC) cells. The proliferation and apoptosis of HNSCC cells treated with the iron chelators DpC and Dp44mT were detected. The mechanism of DpC-induced apoptosis on HNSCC cells was investigated. The human HNSCC cell lines FaDu, Cal-27 and SCC-9 were cultured in vitro and exposed to gradient concentrations of DpC and Dp44mT. A Cell Counting Kit-8 assay was used to detect the viability of FaDu, Cal-27, SCC-9 cells. Double staining with annexin V and propidium iodide was performed for the detection of the proportion of apoptotic FaDu, Cal-27 and SCC-9 cells following treatment. The nuclear damage to Cal-27 cells that were treated with DpC was detected by Hoechst staining. Finally, western blot analysis was used to detect the expression of proteins associated with the DNA damage pathway in Cal-27 cells that were treated with DpC. The CCK-8 assay showed that treatment with DpC and Dp44mT was able to markedly inhibit the viability of FaDu, Cal-27 and SCC-9 cells in a concentration-dependent manner. In comparison to Dp44mT, treatment with DpC exhibited a more effective inhibitory effect on the viability of HNSCC cells. The proportion of apoptotic cells detected by flow cytometry increased in a dose-dependent manner in all cell lines following DpC and Dp44mT treatment, with the proportion of apoptotic HNSCC cells induced by DpC treatment being significantly higher compared with Dp44mT (P<0.05). The results of Hoechst staining revealed that the nuclei of Cal-27 cells exhibited morphological changes in response to DpC treatment, including karyopyknosis and nuclear fragmentation. The expression of DNA damage-associated proteins, including phosphorylated (p)-serine-protein kinase ATM, p-serine/threonine-protein kinase Chk1 (p-Chk-1), p-serine/threonine-protein kinase ATR (p-ATR), p-Chk-2, poly (ADP-ribose) polymerase, p-histone H2AX, breast cancer type 1 susceptibility protein, p-tumor protein P53, increased with increasing concentration of DpC in Cal-27 cells. Treatment with DpC and Dp44mT markedly inhibited cell viability and increased the apoptotic rates in human HNSCC cells in a concentration-dependent manner. DpC exhibited a stronger antitumor effect compared with Dp44mT, potentially inducing the apoptosis of HNSCC cells via the upregulation of DNA damage repair-associated proteins.

Downregulated cytoplasmic polyadenylation element-binding protein-4 is associated with the carcinogenesis of head and neck squamous cell carcinoma.

Cytoplasmic polyadenylation element-binding protein-4 (CPEB4) is involved in several biological processes that are associated with cancer progression. However, it remains unknown whether CPEB4 expression levels are associated with head and neck squamous cell carcinoma (HNSCC). The aim of the present study was to explore the potential function of CPEB4 in HNSCC. The expression of CPEB4 was analyzed in HNSCC from six Gene Expression Omnibus (GEO) datasets. Immunohistochemical staining was conducted to examine CPEB4 protein levels in an HNSCC tissue microarray (TMA). According to the GEO dataset analyses, CPEB4 gene expression was downregulated in HNSCC compared with normal samples (P<0.05). Notably, a statistical difference was observed between different tumor grades (P<0.05). Furthermore, the methylation of the CPEB4 gene in HNSCC was significantly increased compared with that observed in normal samples (P<0.01). The outcome from the TMA demonstrated that CPEB4 protein expression in human HNSCC tumors was significantly decreased compared with normal samples (P<0.05). In addition, the expression of CPEB4 protein was negatively associated with histological grades of HNSCC (P<0.05). The results from the present study suggested that CPEB4 may function as a tumor suppressor gene in HNSCC, which identifies the potential value of CPEB4 in predicting prognosis of HNSCC. Hypermethylation of the CPEB4 gene may be responsible for the downregulation of CPEB4 expression in HNSCC and result in tumorigenesis.

Laminaria Japonica Polysaccharides effectively inhibited the growth of nasopharyngeal carcinoma cells in vivo and in vitro study.

AIMS: Laminaria Japonica Polysaccharides (LJP) is a kind of plant polysaccharide isolated from Laminaria Japonica Aresch. LJP has a variety of biological activity, including anti-tumor, improving immune function, anti-radiation and others. This study observed the biological activity of LJP in vitro and in vivo on human nasopharyngeal carcinoma(NPC), and the possible anticancer mechanism was explored. METHODS: Nasopharyngeal poorly differentiated squamous cell carcinoma cell lines CNE2 and HONE1 were used for the study. MTT method was used to detect the proliferation of HONE1 and CNE2 treated with gradient concentrations of LJP. The apoptosis of HONE1 treated with LJP was detected by annexin V-FITC/PI double staining method. HONE1 was used to establish subcutaneous implanted tumor model in nude mice. The changes of transplanted tumor volume and body weight of nude mice in each group were observed and recorded. The changes of the ultrastructure of transplanted tumor were observed by transmission electron microscope (TEM). RESULTS: MTT results showed that LJP has inhibitory effect on proliferation of both HONE1 and CNE2, and the effects were dosage-dependent; results of flow cytometry (FCM) analysis showed that, LJP could efficient induce apoptosis in HONE1, and apoptosis rate increased with the increase of LJP concentration. In vivo experiments, the inhibition rate was 33.7% (P<0.05) and 47% (P<0.01) in middle and high dose LJP group, respectively. TEM results suggested that the cancer cells in the transplanted tumor tissue treated with middle and high dose LJP presented unique apoptosis changes. CONCLUSIONS: LJP can effectively inhibit the growth of NPC cells. And it may be achieved by inducing apoptosis of NPC cells.

Effect of intraoperative neuromonitoring on recurrent laryngeal nerve palsy rates after thyroid surgery--a meta-analysis.

BACKGROUND/PURPOSE: Though intraoperative nerve monitoring (IONM) during thyroid surgery has gained universal acceptance for localizing and identifying the recurrent laryngeal nerve (RLN), its role in reducing the rate of RLN injury remains controversial. In order to assess the effect of IONM during thyroid surgery, its value in reducing the incidence of RLN palsy was systematically evaluated. METHODS: Studies were evaluated for inclusion in this analysis by researching PubMed, Embase, the Cochrane Central Register of Controlled Trials, and the references of included studies. The initial screening of article titles and abstracts was independently performed by five reviewers based on the research protocol criteria. Each article was then read in detail and discussed before inclusion in the meta-analysis. Data were independently extracted, including the level of evidence, number of at-risk nerves, allocation method, baseline equivalence between groups, definitions of transient and permanent vocal fold palsy, systematic application of electrodes, etc. The meta-analysis was then performed. Odds ratios were pooled using a random effects model. RESULTS: Five randomized clinical trials and 12 comparative trials evaluating 36,487 at-risk nerves were included. Statistically significant differences in terms of total recurrent laryngeal nerve palsy (3.37% with intraoperative nerve monitoring [IONM] vs. 3.76% without IONM [OR: 0.74; 95% confidence interval [CI]: 0.59-0.92]) and transient recurrent laryngeal nerve palsy (2.56% with IONM vs. 2.71% without IONM [OR: 0.80; 95% CI: 0.65-0.99]) were identified. The persistent incidence of recurrent laryngeal nerve palsy was 0.78% for IONM versus 0.96% for nerve identification alone (OR: 0.80; 95% CI: 0.62-1.03). CONCLUSION: Based on this meta-analysis, statistically significant differences were determined in terms of the incidences of total and transient recurrent laryngeal nerve palsy after using IONM versus recurrent laryngeal nerve identification alone during thyroidectomy. However, no statistically significant differences were identified regarding the incidence of persistent recurrent laryngeal nerve palsy between groups.

[The effects of tea polyphenols and Laminaria japonica polysaccharides on nasopharyngeal carcinoma cell HONE1 and CNE2].

OBJECTIVE: To explore the effects of laminaria japonica polysaccharides(LJP) and tea polyphenols (TP) on nasopharyngeal carcinoma(NPC) cells HONE1 and CNE2, and further to explore the underlying mechanism of antitumor activity of LJP on NPC cell in vivo. METHOD: To identify the logarithmic growth phase of NPC cells HONE1 and CNE2 through cell growth curve and doubling time by means of MTT, then inhibition of the cells proliferation were detected with LJP and TP separately and combined. With LJP treatment, cell apoptosis of HONE1 was examined by double staining assay. A tumor model,established by subcutaneously inoculation of NPC cell HONE1 into nude mice,was used to evaluate the inhibitory effect of LJP in vivo. RESULT: Both LJP and TP had inhibition effect on two groups of cell proliferation, and LJP and TP combined effect of inhibition were higher than the two separate on two sets of experimental cell proliferation, whose effect was concentration-dependent. LJP could induce apoptosis of HONE1. With the increasing concentration of LJP, apoptosis rate increased. The apoptosis rate was(49.51 +/- 1. 89) % (P<0. 01) when treated with 320 mg/L LJP. The inhibition rate was between 50% to 60% at 72 h after treatment with 320 mg/L LJP. Compared to control group, the growth of xenografts in nude mice was significantly suppressed after administration of LJP at a dose-dependent manner. The inhibition rates were 33. 7%(P<0. 05)and 47. 0%(P<0. 01) when treated with 25.0 mg/kg and 50. O mg/kg respectively. Whereas the inhibition rate of 12.5 mg/kg group was only 16. 4%(P>0. 05). CONCLUSION: LJP and TP can inhibit the proliferation of NPC cells HONE1 and CNE2 respectively,and combined use has a significant effect. LJP can inhibit the growth of NPC probably by inducing apoptosis of NPC cells in vitro and in vivo.