Integrating iron metabolism-related gene signature to evaluate prognosis and immune infiltration in nasopharyngeal carcinoma.

BACKGROUND: Dysregulation of iron metabolism has been shown to have significant implications for cancer development. We aimed to investigate the prognostic and immunological significance of iron metabolism-related genes (IMRGs) in nasopharyngeal carcinoma (NPC). METHODS: Multiple Gene Expression Omnibus (GEO) and The Cancer Genome Atlas (TCGA) datasets were analyzed to identify key IMRGs associated with prognosis. Additionally, the immunological significance of IMRGs was explored. RESULTS: A novel risk model was established using the LASSO regression algorithm, incorporating three genes (TFRC, SLC39A14, and ATP6V0D1).This model categorized patients into low and high-risk groups, and Kaplan-Meier analysis revealed significantly shorter progression-free survival for the high-risk group (P < 0.0001). The prognostic model's accuracy was additionally confirmed by employing time-dependent Receiver Operating Characteristic (ROC) curves and conducting Decision Curve Analysis (DCA). High-risk patients were found to correlate with advanced clinical stages, specific tumor microenvironment subtypes, and distinct morphologies. ESTIMATE analysis demonstrated a significant inverse relationship between increased immune, stromal, and ESTIMATE scores and lowered risk score. Immune analysis indicated a negative correlation between high-risk score and the abundance of most tumor-infiltrating immune cells, including dendritic cells, CD8+ T cells, CD4+ T cells, and B cells. This correlation extended to immune checkpoint genes such as PDCD1, CTLA4, TIGIT, LAG3, and BTLA. The protein expression patterns of selected genes in clinical NPC samples were validated through immunohistochemistry. CONCLUSION: This study presents a prognostic model utilizing IMRGs in NPC, which could assist in assessing patient prognosis and provide insights into new therapeutic targets for NPC.

Follistatin-Like Protein-1 Upregulates Dendritic Cell-Based Immunity in Patients with Nasopharyngeal Carcinoma.

Follistatin-like protein-1 (FSTL1) is an inflammatory factor that can induce an inflammatory response and is expressed in cancers. However, little is known about its content and function in nasopharyngeal carcinoma (NPC). Interleukin (IL)-12 and IL-4 are primarily secreted by dendritic cells (DCs) and activated T lymphocytes, respectively; these factors can induce Th cell differentiation and cytotoxic lymphocyte production, both of which facilitate tumors through the STAT4 and STAT6 pathways, respectively. In this study, the relationship between FSTL1 and both IL-12 and IL-4 as well as the functional mechanism of these cytokines was explored. Enzyme-linked immunosorbent assay, flow cytometry, and Western blotting were used to assess the levels of key inflammatory factors and DC markers as well as elucidate the mechanism by which FSTL-1 mediates and exerts it antitumor effects. The results revealed that serum FSTL1 and IL-12 levels were significantly decreased in NPC patients compared with those in the control group (P < 0.05); conversely, IL-4 levels were increased (P < 0.05). Supernatants from the experimental groups (EGs) contained higher IL-4 and IL-12 levels than those from the control groups (P < 0.05). Additionally, phosphorylated-STAT6 and phosphorylated-STAT4 were increased in the EGs (P < 0.05). These results suggest that DC-mediated immunity was activated by FSTL1, which leads to an increase of IL-12 and IL-4 production and consequently activates the STAT4 and STAT6 pathways through upregulation of STAT4 and STAT6 phosphorylation, respectively.

Follistatin-like protein 1 contributes to dendritic cell and T-lymphocyte activation in nasopharyngeal carcinoma patients by altering nuclear factor κb and Jun N-terminal kinase expression.

Follistatin-like protein 1 (FSTL1) is a newly characterized protein that can regulate the immune response in various ways. Dendritic cells (DCs) are central to immune regulation. In this study, we explored the impact of FSTL1 on DC activity in nasopharyngeal carcinoma (NPC) patients. The surface expression of CD40, CD86, and HLA-DR on DCs was analyzed and showed significantly elevated expression levels, indicating DC maturity. After FSTL1 was added to DCs collected from NPC patients (n = 50), controls (n = 47), and healthy donors (n = 10), interferon γ secretion and T-cell receptor expression in cytotoxic T lymphocytes were also investigated. In the experimental groups, the expression of the critical immune protein nuclear factor (NF)-κb was upregulated, whereas Jun N-terminal kinase (JNK) was downregulated. Our findings demonstrate that FSTL1 plays a critical role in immune regulation, enhancing the antigen presentation ability of DCs by up-regulating NF-κb expression and down-regulating JNK expression.

Epigenetic inactivation of follistatin-like 1 mediates tumor immune evasion in nasopharyngeal carcinoma.

Follistatin like-1 (FSTL1) is a secreted glycoprotein involved in a series of physiological and pathological processes. However, its contribution to the development of cancer, especially the pathogenesis of nasopharyngeal carcinoma (NPC), remains to be elucidated. We aimed to investigate the dysregulation of FSTL1 and its possible function in NPC. FSTL1 was frequently downregulated in NPC cell lines and primary tumor biopsies by promoter hypermethylation. Ectopic expression of FSTL1 significantly suppressed the colony formation, proliferation, migration and invasion ability of NPC cells and induced cell apoptosis. Overexpression of FSTL1 decreased the tumorigenicity of NPC cells in vivo. In addition, the proliferation of NPC cells in vitro was inhibited by treatment with soluble recombinant FSTL1 protein. The protein level of FSTL1 was decreased in primary NPC tumors and was associated with downregulated interleukin 1ß (IL-1ß) and tumor necrosis factor α (TNF-α). Furthermore, recombinant human FSTL1 protein induced secretion of IL-1ß and TNF-α in macrophage cultures, therefore FSTL1 might activate macrophages and attenuate the immune evasion of NPC cells. In conclusion, the epigenetic downregulation of FSTL1 may suppress the proliferation and migration of NPC cells, leading to dysfunctional innate responses in surrounding macrophages.

Tea polyphenols protect against irradiation-induced injury in submandibular glands' cells: a preliminary study.

AIM: To study the protective effect of tea polyphenols (TPs) on submandibular glands affected by radiation injury. METHODS: Sixty rats were randomly divided into radiation group (R-group, N = 30) and TP-pre-treated-radiation group (TPR-group, N = 30). The rats were intragastrically administered with TP or normal sodium from 14 days before radiation, continuously daily, until the experiment. All the rats in both groups were irradiated with a single exposure dose of 15 Gy gamma rays that were delivered to the head and neck areas. Ten rats of each group were anatomised on the 3rd, 6th and 30th day after irradiation, respectively. The submandibular glands of the rats were removed for the study. The morphologic changes of the submandibular glands were observed by transmission electron microscopy (TEM). The terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate (dUTP)-biotin nick-end labelling (TUNEL) method was used to detect apoptosis of the submandibular glands' cells. RESULTS: Electron microscope observation of the submandibular glands showed that the lesions of the TPR-group were mild. Change in apoptosis of the cells was not obvious compared with the R-group. The cell apotosis was typical after irradiation in the R-group. Apoptosis index that was detected in the cells of submandibular glands of the TPR-group was statistically significantly decreased compared with the R-group (P < 0.01) on the 3rd, 6th and 30th day after irradiation. CONCLUSION: TP could protect submandibular glands from radiation injuries, and the protection mechanism may be realised by anti-apoptosis.

[Expression and relationship of EBV LMP1, COX-2 and VEGF-C in nasopharyngeal carcinoma].

OBJECTIVE: To evaluate the expression of vascular endothelial growth factor C (VEGF-C), Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1), cyclooxygenase-2 (COX-2) in nasopharyngeal carcinoma (NPC). METHOD: LMP1, COX-2 and VEGF-C were detects by immunohistochemical staining for 57 case NPC tissue. RESULT: The positives rates of LMP1, COX-2 and VEGF-C detected by immunohistochemical staining were 49.1% (28/57), 75.4% (43/57) and 59.6% (34/57), respectively. The expression of LMP1, COX-2 and VEGF-C were correlated to each other in NPC (P < 0.05). CONCLUSION: LMP1 and COX-2 may induce expression of VEGF-C directly or LMP1 induce expression of VEGF-C by induce COX-2 expression, may contribute to lymph metastasis and develop NPC.

Effect of 5-ALA-PDT on VEGF and PCNA expression in human NPC-bearing nude mice.

Photodynamic therapy (PDT) is a promising treatment for nasopharyngeal carcinoma. However, recurrence and metastasis of the tumor after PDT remains problematic. In this study we investigated VEGF and PCNA expression in tumor tissues after 5-ALA-PDT. BALB/c nude mice with NPC tumors of similar size were randomly assigned to three groups (n=10 each). In the two treatment groups, mice were administrated 5-ALA intratumorally at a dose of 100 mg/kg and 3-3.5 h prior to laser irradiation (630 nm, 100 J/cm(2), 100 mW/cm(2)). The mice in one of the treatment groups were sacrificed at 24 h after PDT. The other treatment group and control group mice were sacrificed 14 days after PDT, and the tumor weights were determined for all three groups. Mean tumor weights at 14 days after PDT were 1.353+/-0.204 g in the treatment group and 3.124+/-0.380 g in the control group (p<0.001). Results showed the VEGF level in tumor tissues at 24 h after PDT was slightly higher than that in the control group, while it was down-regulated at 14 days after PDT. The PCNA level was not significantly different in tumor tissues among the three groups, but it was lower in degenerated tumor cells 24 h after PDT. Our results suggest that VEGF may play a role in tumor recurrence and metastasis following PDT. Residual tumor cells escaped from PDT is the main reason for tumor recurrence.

[Relationship between the expression of TrkB and anoikis resistance in nasopharyngeal carcinoma line].

OBJECTIVE: To study the regulation of anoikis by tyrosine kinase receptor B (TrkB) in human nasopharyngeal carcinoma lines. METHODS; Expression levels of TrkB and brain-derived neurotrophic factor (BDNF) were evaluated by RT-PCR and Western blot. Colony formation ability of C666-1 was observed in soft agar. Proliferation rate and apoptosis, that change in cells by treating the TrkB inhibitor K252a and specificity ligand BDNF respectively under suspension culture, were measured by cell counting kit-8 (CCK-8) assay and the flow cytometry assay. The expression change of TrkB, BDNF and phosphorylation of serine threonine kinase (p-Akt) were investigated by Western blot analysis. RESULTS: TrkB and BDNF were identified in C666-1 cells. C666-1 cells could be decreased the proliferation of colony in soft agar by effect of K252a, but BDNF could make the colony prolific. K252a can inhibit the expression of TrkB in C666-1, and prevent p-Akt activation. And exogenous BDNF stimulated up-regulation TrkB and p-Akt, induced anoikis resistance. CONCLUSION: TrkB inhibits anoikis in nasopharyngeal carcinoma cells. Inhibiton of TrkB by K252a can induce anoikis, and may prove particularly effective in treatment of nasopharyngeal carcinoma.

[Expression and significance of tyrosine kinase receptors B in nasopharyngeal carcinoma patients].

OBJECTIVE: This study was to explore the expression and clinicopathologic features of Tyrosine kinase receptors B (TrkB) and its ligand brain-derived neurotrophic factor (BDNF) in nasopharyngeal carcinoma (NPC). METHOD: Immunohistochemistry was adopted to detect the expression level of TrkB and BDNF in NPC patients. RESULT: Both TrkB and BDNF were expressed in NPC as well as in chronic inflammation. The active expression rate of TrkB in NPC was 82.5% (47/57) and BDNF was 52.6% (30/57), both of which were higher than those in chronic inflammation (P < 0.05). The degree of TrkB expression was more marked in T3 + T4, III + IV stage NPC than that in T1 +T2, I + II stage NPC (P < 0.05). TrkB abnormal expression rate of nasopharyngeal carcinoma with lymph node metastasis was higher than that of NPC without lymph node metastasis (P < 0.05). No statistical significance for degrees of TrkB expression in pathologic type grades was found (P > 0.05). There were no statistical significance for degrees of BDNF expression in T stage, clinical stage and lymph node metastasis in NPC (P > 0.05). The expression of TrkB was unrelated to the expression of BDNF (r = 0.049, P > 0.05). CONCLUSION: The high expression rate of TrkB and BDNF maybe plays an important role in development of NPC. It is suggested that TrkB and its ligand BDNF may act as an important index for forecasting the development and metastasis of NPC.

[The analysis of plasma Urokinase-type plasminogen activator and Urokinase-type plasminogen activator receptor which unknown factor nosebleed patients].

OBJECTIVE: To investigate the pathogenesis of unknown nosebleed patients. METHOD: The ELISA test were used to detected plasma Urokinase-type plasminogen activator (uPA) and Urokinase-type plasminogen activator receptor (uPAR) level in 19 cases unknown factor nosebleed patients and 36 health persons. RESULT: The results showed uPAR and uPA level in nosebleed group (before treatment) uPAR (0.14 +/- 0.04) microg/L, uPA (0.24 +/- 0.09) microg/L; (after treatment) uPAR (0.08 +/- 0.02) microg/L, uPA (0.18 +/- 0.07) microg/L. And normal group uPAR (0.07 +/- 0.03) microg/L, uPA (0.17 +/- 0.05) microg/L. The uPAR and uPA level in nosebleed group before treatment is higher than that in normal group (P <0.05). There is no significant difference between nosebleed group after treatment and normal group (P>0.05). CONCLUSION: The reasons of uPAR and uPA level high in unknown factor nosebleed patients were not clear, maybe relation to vascular endothelial cell, smooth muscle cell and neutrophil-monocytic release more uPAR and uPA. So uPAR and uPA density of nostril accumulation is more high in its microenvironment, that fibrinolytic system activated increase and result in its hyperactivity, and happened nosebleed when blood be in hypocoagulable state.

[Application of EB virus latent membrane protein 1 in nasopharyngeal swab in diagnosis of nasopharyngeal carcinoma].

OBJECTIVE: To investigate the feasibility and reliability of detection of Epstein-Barr virus (EBV) latent membrane protein-1 (LMP1) gene by nasopharyngeal swab in the diagnosis of nasopharyngeal carcinoma (NPC). To investigate the distribution of 30 bp deletion variant of LMP-1 gene in the local population. METHOD: Nasopharyngeal cells were collected by nasopharyngeal swab, and then DNA was extracted, which was subsequently confirmed by amplification of sequence of beta-thalassemia gene by polymerase chain reaction (PCR). And sequence of LMP1 was amplified with specific primer to verify the significance of LMP1 in the diagnosis of NPC. RESULT: DNA was obtained from 96.4% nasopharyngeal swab samples, LMP1 was detected in 33 of 36 samples, while 2 of 45 samples from normal control, with sensitivity 91.7%, and specialty 95.6%. 30 bp deletion of LMP1 gene was found in 80.6% of NPC samples, and wild type 11.1%. CONCLUSION: Our study suggests that nasopharyngeal swab could be effective method for gene-detection. As a parameter in diagnosis of NPC, LMP1 gene is maybe superior to EBVCA-IgA. 30 bp deletion of LMP1 oncogene is widespread in NPC patients.

[A study of biological characteristic and generation of monocyte-derived dendritic cell from peripheral blood of patients with nasopharyngeal carcinoma].

OBJECTIVE: To study the differences of morphology, phenotype and function of dendritic cell (DC) derived from the healthy individuals and the patients with nasopharyngeal carcinoma (NPC). METHOD: DC were isolated and developed from peripheral blood monocytes from 12 NPC patients and 9 healthy volunteers in vitro, and were studied in terms of shape and phenotype of CD83, CD1a,CD86, HLA-DR by flow cytometry then. MTT method was used to measured the capability of DC in mixed lymphocyte reactions (MLR). RESULT: The shape of DC derived from monocyte in the healthy volunteers' peripheral blood was more typical than that of NPC patients'. The expression rate of CD83, CD1a on DC from healthy volunteers was significantly higher than that from NPC patients (P < 0.05). The DC from healthy volunteers had the higher expression level of CD1a and CD83 than that from NPC patients, but not significantly( P > 0.05). DC in healthy volunteer's group could trigger proliferation of allogenous lymphocyte (P < 0.05) but it could not significantly do that in NPC patients group (P > 0.05). CONCLUSION: These results reveal that the yield of DC derived from NPC patient's monocyte is lower than that from healthy individual's in peripheral blood,and the DC from NPC patient may be in an immature state and could not trigger the proliferation of lymphocyte.

[Analysis of the nasal complication after radiotherapy in patients with nasopharyngeal carcinoma by logistic regression].

OBJECTIVE: To study of the risk factors of the nasal complication after radiotherapy in patients with nasopharyngeal carcinoma(NPC). METHOD: Observed 131 cases nasal disease in NPC patients, analyzed the factors that might cause the complication by using Logistic regression model. RESULT: The highest incidences of complication after radiotherapy was chronic purulent sinusitis (116/131), the chosen factors in the regression formula were T staging and radiation for anterior field of the nose. The second was nasal synechia(33/131), the chosen factors were total dose of external irradiation, intracavitary irradiation and medicamentous rhinitis. The incidences of atrophic rhinitis was the lowest (9/131), the chosen factors were chemotherapy and medicamentous rhinitis. CONCLUSION: T staging of tumor and radiation for anterior field of the nose had relation with post radiotherapy sinusitis. It was easy to lead to nasal synechia when the does of external irradiation being higher or adding intracavitary irradiation or using the decongestant for long time. Chemotherapeutic drugs and decongestant may be the risk factors of atrophic rhinitis.

[Clinical and pathological study of chordoma in the skull base].

OBJECTIVE: To study the clinical and pathological characteristics of chordoma in the skull base. METHOD: The clinical manifestation, radiological finding and pathological characteristics in 20 patients with skull base chordoma were reviewed. Among them, 7 were immunohistochemical stained for keratin, epithelial membrane antigen, vimentin, glial fibrillary acidic protein and desmen, 2 were studied by electron microscope. 19 patients received surgical resection(5 cases were assisted by nasal endoscope). Eleven patients received radiation therapy. All 19 patients were followed-up for 3-5 years after treatment. RESULTS: (1) Clinical type were divided into four types in this group according to symptoms, sign, imaging features, operation finding, the original places and the expansive direction of the tumor. Seller type 5 cases, clival type 9 cases, occipito-temproral type 2 cases and extent type 4 cases. One case with multiple cranial never palsies died due to the pressure on brain stem by the neoplasm, was autopsied prove lungs metastasis. 5 years survival rates were 65% and 35% respectively. (2) Vacuolated physaliferous cell is the pathology characteristic of chordoma. (3) Positive rate of epithelial marker antigens are higher than other mesenchymal neoplasms. Distended rough endoplasmic reticulum and filament were found within the cytoplasm under transmission electron microscope. CONCLUSION: Chordoma is a kind of low malignancy which express epithelial and mesenchymal characteristics. Vacuolated physaliferous cell is the characteristic Pathologic appearance. Immunohistiochemical stain is helpful in differential diagnosis. Complete excision is still technically impossible. Chordoma maintain high recurrence rate after surgery, with metastasis. Combined modality therapy include surgical operation and radiation could prolonged survival period.

[Preservation of laryngeal and glossal function in surgical treatment of oropharyngeal glossal epiglottic carcinoma].

OBJECTIVE: To study the feasibility, surgical techniques and results of surgical treatment of oropharyngeal glossal epiglottic carcinoma with laryngeal and glossal functions preserved. METHODS: A retrospective review of 21 (17 males, 4 females, age range 44 to 72 years) cases with carcinoma extending from the oropharynx, glossa to the epiglottis treated during 1990 to 2000 was conducted. The clinical stage was T2N0 in 4, T2N1 in 5 and T3N1 in 12 according to 1992 UICC. All the cases were undergone partial resection of the glossal base-epiglottis and suproglottic horizontal partial laryngectomy. Seventeen cases received selective neck dissections and all the patients received postoperative radiotherapy (50-60 Gy). RESULTS: The 3-year and 5-year survival rates in this series were 73.3% and 57.1%, respectively. Decannulation and complete preservation of laryngeal and glossal functions were achieved in 20 cases (95.7%), while partial preservation (speaking and deglutition) in one case (4.2%). CONCLUSION: Preservation of the laryngeal and glossal functions in surgical treatment of oropharyngeal glossal epiglottic carcinoma is feasible. Subhyoid approach was proved to be the best way for tumor resection in such an extent.