Metabolic stress-induced reciprocal loop of long noncoding RNA ZFAS1 and ZEB1 promotes epithelial-mesenchymal transition and metastasis of pancreatic cancer cells.

Pancreatic cancer (PC) development faces significant metabolic stress due to metabolic reprogramming and a distinct hypovascular nature, often leading to glucose and glutamine depletion. However, the adaption mechanisms by which PC adapts to these metabolic challenges have not yet been completely explored. Here, we found that metabolic stress induced by glucose and glutamine deprivation led to an overexpression of ZNFX1 antisense RNA 1 (ZFAS1). This overexpression played a significant role in instigating PC cell epithelial-mesenchymal transition (EMT) and metastasis. Mechanistically, ZFAS1 enhanced the interaction between AMPK, a key kinase, and ZEB1, the primary regulator of EMT. This interaction resulted in the phosphorylation and subsequent stabilization of ZEB1. Interestingly, ZEB1 also reciprocally influenced the transcription of ZFAS1 by binding to its promoter. Furthermore, when ZFAS1 was depleted, the nutrient deprivation-induced EMT of PC cells and lung metastasis in nude mice were significantly inhibited. Our investigations also revealed that ZFAS1-rich exosomes released from cells suffering glucose and glutamine deprivation promoted the EMT and metastasis of recipient PC cells. Corroborating these findings, a correlated upregulation of ZFAS1 and ZEB1 expression was observed in PC tissues and was associated with a poor overall survival rate for patients. Our findings highlight the involvement of a long noncoding RNA-driven metabolic adaptation in promoting EMT and metastasis of PC, suggesting ZFAS1 as a promising novel therapeutic target for PC metabolic treatment.

Effects of modified BOPPPS-based SPOC and Flipped class on 5th-year undergraduate oral histopathology learning in China during COVID-19.

BACKGROUND: Colleges and universities in China have offered courses based on online teaching platforms as required by the Ministry of Education since the beginning of the COVID-19 pandemic. This emergency action was not an expedient measure, but a powerful impetus to improve extant education and implement teaching reform. Oral histopathology is a basic subject in oral medicine education, which combines theory with practice. The course aims to improve the ability of students to observe, think, analyze and identify oral diseases. METHOD: We adjusted and modified the original Bridge-In, Outcomes, Pre-assessment, Participatory Learning, Post-assessment, and Summary (BOPPPS) teaching method to fit the characteristics and needs of oral histopathology. We then combined the characteristics of Small Private Online Courses (SPOCs) and a Flipped class to complete teaching material online, and assessed the effects of such teaching using a questionnaire and interviews. Fifty 5th-year undergraduates in stomatology at the School of Stomatology of Harbin Medical University of China participated in online classes. All were in the junior second half of the semester at the beginning of 2020. Teachers investigated from various medical colleges were responsible for delivering courses associated with stomatology or ophthalmology. RESULT & CONCLUSION: The results showed that the modified BOPPPS combined with SPOC and the Flipped class improved teaching satisfaction. Modified BOPPPS combined with SPOC and the Flipped class is a useful complement to offline teaching on 5th-year undergraduate oral histopathology learning in China during COVID-19, and it can meet the multiple needs of students participating in the course.

Targeting IL-17alpha to promote anti-PD-1 therapy effect by screening the tumor immune microenvironment in a mouse oral carcinogenesis model.

BACKGROUND: Resistance to PD-1 blocking agents is not uncommon, limiting their wide clinical success. Certain tumor-infiltrating immune cells (e.g., TILs/CTLs) have emerged as biomarkers of response, and absence of such immune cells contributes to resistance. OBJECTIVE: We deconvoluted the dynamic immune microenvironment in a mouse model of oral carcinogenesis for augmenting the resistance to PD-1 blocking agents by combination. METHODS: Bioinformatics methods and routine biological experiments were adopted such as morphological analysis and ELISA in the 4NQO-treated mice model. RESULTS: Our findings revealed that dysplastic tongue tissues from 4NQO-treated mice were characterized by an immunosuppressive tumor microenvironment. Tongue tissues from mice treated with 4NQO for 12 weeks had higher levels of Th2 cells and Tregs compared to tissues taken from control mice or mice treated with 4NQO for 28 weeks; these results suggested a potential therapeutic benefit of anti-PD-1 in the oral cancer. The IL-17 pathway was significantly upregulated during progression from normal mucosa to hyperplasia and tumor formation in mice. Inhibition of IL-17α combined with PD-1 blockade delayed the development of 4NQO-induced precancerous and cancerous lesions and prolonged the survival of 4NQO-treated mice. CONCLUSIONS: Our data suggested a strong rationale of IL-17α blockade as a potential approach to augment the tumor-eliminating effects of anti-PD-1 therapy.

HMGB1-induced autophagy promotes extracellular matrix degradation leading to intervertebral disc degeneration.

Intervertebral disc degeneration (IDD) remains a leading cause of adult disability. High mobility group box l (HMGB1) is a nuclear DNA-binding protein and acts as a central mediator of inflammation. The purpose of this study was to investigate the effect of HMGB1 in IDD. In our study, IDD intervertebral disc tissues were collected and nucleus pulposus cells (NPCs) were primarily cultured. The HMGB1 expression and the effect of HMGB1 on NPCs and extracellular matrix and autophagy were all evaluated. Results showed that HMGB1 was markedly overexpressed in IDD (P<0.05), and upregulated expression of HMGB1 can inhibit NPC proliferation and promote NPC apoptosis (P<0.05), promote extracellular matrix degradation, and activate cell autophagy (P<0.05). Therefore, we concluded that HMGB1 was up-regulated in IDD and HMGB1-induced autophagy can promotes extracellular matrix degradation and thus lead to intervertebral disc degeneration. In brief, HMGB1 may serve as a novel diagnostic biomarker and therapeutic target for IDD.

Impact of immunodeficiency on lingual carcinogenesis and lymph node metastasis in mice.

PURPOSE: Carcinogenesis is mostly accompanied by inflammation and is shaped by immune responses, where the host immune system does not always appear to be a powerful active tool. Some studies have identified that T lymphocytes in the tumor microenvironment are indispensable for the process of distant metastasis. This study aimed to investigate the contradictory role of the immune system and to examine the impact of immunodeficiency on carcinogenesis and lymph node metastasis in mouse models. METHODS: BALB/c mice, BALB/c nude mice, and CB-17SCID mice were treated with 50 mg/L 4-nitroquinoline-1-oxide (4NQO) in drinking water for 16 weeks followed by distilled water until 38 weeks. Three mice per group were sacrificed biweekly or every four weeks from week 4, and tongue lesions and regional lymph node metastases were evaluated. RESULTS: There were no significant differences in the histological examination of carcinogenesis in immunocompetent and immunodeficient mice exposed to 4NQO. However, development of cancer in immunodeficient mice (12 weeks) was earlier than that in immunocompetent mice (28 weeks). Local lymph metastases were observed after 36 weeks in immunocompetent BALB/c mice as opposed to after 18 weeks in immunodeficient mice. CONCLUSION: Immunodeficiency could remarkably advance carcinogenic progression and lymph node metastasis but might not accelerate lymph node metastasis progression.

Prognostic implication of NOTCH1 in early stage oral squamous cell cancer with occult metastases.

OBJECTIVE: The objective of this study was to explore the prognostic value of cancer stem cell markers, namely CD133, NANOG, and NOTCH1, in early stage oral squamous cell carcinoma (OSCC). MATERIALS AND METHODS: One hundred forty-four patients with early stage (cT1T2N0) OSCC were identified from a pre-existing database of patients with oral cancer. We examined the impact of the immunohistochemical expression of CD133, NANOG, and NOTCH1 in OSCC. Overall survival (OS) curves were calculated using the Kaplan-Meier method. Predictors of outcome were identified using multivariate analysis. RESULTS: We found that CD133, NANOG, and NOTCH1 were significantly associated with lymph node metastasis, and NOTCH1 was also significantly associated with depth of invasion and locoregional recurrence. CONCLUSIONS: NOTCH1 was identified as an independent prognostic factor for OS. CLINICAL RELEVANCE: NOTCH1 might prove to be a useful indicator for high-risk patients with occult metastases from early stage OSCC.

A pilot study: Screening target miRNAs in tissue of nonsyndromic cleft lip with or without cleft palate.

Nonsyndromic cleft lip with or without cleft palate (NSCLP) has been recognized as a condition resulting from a combination of environmental and genetic factors. Studies have demonstrated that microRNAs (miRNAs) are involved in embryonic development. However, few studies have focused on screening potential target miRNAs in human NSCLP tissue. Using microarray-based miRNA expression profiling, miRNA expression was compared in tissue samples from 4 NSCLP patients and 4 healthy control subjects. Two hundred and fifty-four miRNAs were found to be differentially expressed. Changes in Homo sapiens (hsa)-miR-24-3p, hsa-miR-27b-3p, hsa-miR-205-5p, hsa-miR-1260b and hsa-miR-720 were of particular interest with respect to Wnt signaling (fold-changes were 12.5, 12.2, 12.1, 12.3 and 10.5, respectively; P<0.005 for all). The levels of hsa-miR-24-3p, hsa-miR-1260b and hsa-miR-205-5p were higher in tissues from NSCLP patients than in those from controls according to PCR analysis. Hsa-miR-24-3p, hsa-miR-1260b and hsa-miR-205-5p may be candidate miRNAs involved in the etiology of NSCLP via Wnt signaling.

Telmisartan suppresses cardiac hypertrophy by inhibiting cardiomyocyte apoptosis via the NFAT/ANP/BNP signaling pathway.

Telmisartan, a type of angiotensin II (Ang II) receptor inhibitor, is a common agent used to treat hypertension in the clinic. Hypertension increases cardiac afterload and promotes cardiac hypertrophy. However, the ventricular Ang II receptor may be activated in the absence of hypertension. Therefore, telmisartan may reduce cardiac hypertrophy by indirectly ameliorating hypertensive symptoms and directly inhibiting the cardiac Ang II receptor. Nuclear factor of activated T­cells (NFAT) contributes to cardiac hypertrophy via nuclear translocation, which induces a cascade of atrial natriuretic peptide (ANP) and brain/B­type natriuretic peptide (BNP) expression and cardiomyocyte apoptosis. However, NFAT-mediated inhibition of cardiac hypertrophy by telmisartan remains poorly understood. The present study demonstrated that telmisartan suppressed cardiomyocyte hypertrophy in a mouse model of cardiac afterload and in cultured cardiomyocytes by inhibiting NFAT nuclear translocation, as well as by inhibiting ANP and BNP expression and cardiomyocyte apoptosis, in a dose­dependent manner. The present study provides a novel insight into the potential underlying mechanisms of telmisartan-induced inhibition of cardiomyocyte hypertrophy, which involves inhibition of NFAT activation, nuclear translocation and the ANP/BNP cascade.

In situ memory T cells and patterns of invasion predict outcome in patients with early-stage oral squamous cell carcinoma.

PURPOSE: The objective of this study was to explore the prognostic value of in situ memory T cells and patterns of invasion (POI) in early stage oral squamous cell carcinoma (OSCC). METHODS: One hundred fifty-seven patients with early stage (cT1T2N0) OSCC were identified from a pre-existing database of patients with oral cancer and were classified by POI according to hematoxylin-eosin staining. We examined the impact of the immunohistochemical expression of CD45RO in OSCC. Overall survival (OS) curves were calculated using the Kaplan-Meier method. RESULTS: Margin status was significantly associated with local recurrence in early stage OSCC (p= 0.000), and depth of invasion was also associated with regional recurrence (2 mm: p= 0.034; 4 mm: p= 0.015). The expression of CD45RO (p= 0.021) and POI (p= 0.027) individually was associated with OS, and patients in the group with combination score tended to have worse OS (p= 0.012). CONCLUSION: Combined evaluation of POI and CD45RO might prove to be a useful indicator for high-risk patients with occult metastases from early stage OSCC.

Combined Expression of c-jun, c-fos, and p53 Improves Estimation of Prognosis in Oral Squamous Cell Carcinoma.

To identify the prognostic value of c-jun, c-fos, and p53 in oral cancer, we examined the impact of immunohistochemical expression of these markers on tumor progression in 157 oral squamous cell carcinoma (OSCC). We found that c-jun or c-fos was significantly associated with lymph node metastasis, and coexpression of c-jun/c-fos, or c-jun/c-fos/p53 were significantly associated with lymph node metastasis, poor differentiation and clinical stage. The coexpression of c-jun/c-fos/p53 was identified as independent prognostic factors for overall survival. Simultaneous coexpression of these markers in OSCCs might prove to be a useful indicator for differentiation of low and high-risk patients.

Expression of CD163, interleukin-10, and interferon-gamma in oral squamous cell carcinoma: mutual relationships and prognostic implications.

Tumor-associated macrophages (TAMs) and their associated inflammatory cytokines represent the major inflammatory component of the stroma of many tumors and can affect prognosis in the case of neoplasms. The objective of this study was to determine the prognostic significance of CD163(+) cells, interleukin-10 (IL-10), and interferon-gamma (IFN-γ) in oral lesions associated with oral squamous cell carcinoma (OSCC). The levels of CD163, IFN-γ, and IL-10 in the tissue samples of 240 patients with OSCC and 58 patients with other oral lesions were assessed by immunohistochemistry. Individuals with low IFN-γ levels, high IL-10 levels, and low CD163 levels were of special concern with respect to OSCC progression. We found that high levels of CD163, or a combination of low IFN-γ levels, high IL-10 levels, and low CD163 levels, were associated with poorer overall survival (OS). CD163(+) cells provide better predictive power for OS in comparison with traditional markers, such as clinical stage and lymph node metastasis. Therefore, CD163(+) cells may be effective prognostic predictors of OSCC. IL-10 may also indicate poor outcomes when IFN-γ secretion is low and the cells are CD163(-) .

Paxillin expression levels are correlated with clinical stage and metastasis in salivary adenoid cystic carcinoma.

AIMS: To investigate the relationship between paxillin expression and clinicopathological features and metastasis in salivary adenoid cystic carcinoma (SACC). METHODS: A total of 47 SACC were assessed histochemically for paxillin expression. Paxillin immunoreactivity was compared with histological type, clinical stage and distant metastasis. RESULTS: Paxillin expression was identified in 57.45% of SACC as cytoplasmic staining and the expression was correlated with distant metastasis and clinical stage (P < 0.05), but not with histological type. CONCLUSIONS: Our observations indicate that paxillin expression is upregulated in SACC. High expression of paxillin was correlated with a more advanced stage and metastasis in SACC, suggesting that paxillin is a disease marker in advanced SACC and SACC with distant metastasis, and, consequently, may have value as a therapeutic target for SACC.

Detection of deregulated pathways to lymphatic metastasis in oral squamous cell carcinoma.

Oral squamous cell carcinoma (OSCC) is a common malignancy, in which lymph node metastasis is a major determinant of outcome. The pathway deregulation resulting from a large number of somatic genetic alterations in the development of the tumor, plays an important role in lymphatic metastasis process. To detect the deregulated pathways to lymphatic metastasis in OSCC, we performed pathway-oriented analysis using gene expression profile from 16 samples without lymphatic metastasis and 27 samples with lymphatic metastasis. We identified seven significantly (p < 0.05) deregulated pathways: the erythropoietin (EPO) Signaling Pathway, Signaling Pathway from G-Protein Families, Cytokine-cytokine receptor interaction, the Janus kinase (JAK)-signal transducer and activator of transcription (STAT) signaling pathway, Ribosome, Colorectal cancer, B cell receptor signaling pathway. The biological relevance of these pathways to OSCC is the focus of ongoing studies, as well as complex interactions and crosstalk between them. These pathways might provide additional clues about factors that regulate the course for OSCC patients and might offer new opportunities for therapeutic intervention.