Constructing a Ferroptosis-related Long Non-coding RNA Signature to Predict the Prognostic of Head and Neck Squamous Cell Carcinoma Patients by Bioinformatic Analysis.

Ferroptosis is a novel discovered iron-dependent mode of regulated cell death (RCD) which characterized non-apoptosis. Researches have shown the effect of ferroptosis in the biological activities of tumors. But there is no relevant study showing the relationship between ferroptosis-related genes and head and neck squamous cell carcinoma (HNSCC). In this study, we paid attention to several ferroptosis-related lncRNAs in head and neck squamous cell carcinoma and its clinical features. HNSCC data were obtained from The Cancer Genome Atlas (TCGA) database. Ferroptosis-related lncRNAs were selected by the coexpression network. Then, we identified prognostic ferroptosis-related lncRNAs in HNSCC patients and constructed a ferroptosis-related lncRNAs signature by Lasso cox regression. Besides, GSEA analysis was performed to explore the functional enrichment of ferroptosis-related lncRNAs. Nine ferroptosis-related lncRNAs (AC004687.1, AL450992.2, AC010894.2, AL451085.2, AC104083.1, LIPE-AS1, AC108010.1, CTBP1-DT, and PTCSC2) were identified to have the independent prognostic value in HNSCC patients and the ferroptosis-related lncRNAs signature was constructed based on these nine genes. According to the risk score of the signature, the high-risk group had shorter overall survival (OS) compared with the low-risk group. Risk score showed to be an independent factor for the patients with HNSCC. Additionally, the nomogram was generated and the nine lncRNAs were mainly enriched in phagocytosis, metabolism, and chemokine signaling pathways. The ferroptosis-related lncRNA signature has effects on the prognostic prediction of patients with HNSCC which may serve as potential therapeutic targets for patients with HNSCC.

Picropodophyllin inhibits tumor growth of human nasopharyngeal carcinoma in a mouse model.

Insulin-like growth factor-1 receptor (IGF-1R) is a cell membrane receptor with tyrosine kinase activity and plays important roles in cell transformation, tumor growth, tumor invasion, and metastasis. Picropodophyllin (PPP) is a selective IGF-1R inhibitor and shows promising antitumor effects for several human cancers. However, its antitumor effects in nasopharyngeal carcinoma (NPC) remain unclear. The purpose of this study is to investigate the antitumor activity of PPP in NPC using in vitro cell culture and in vivo animal model. We found that PPP dose-dependently decreased the IGF-induced phosphorylation and activity of IGF-1R and consequently reduced the phosphorylation of Akt, one downstream target of IGF-1R. In addition, PPP inhibited NPC cell proliferation in vitro. The half maximal inhibitory concentration (IC50) of PPP for NPC cell line CNE-2 was ≤1 µM at 24h after treatment and ≤0.5 µM at 48 h after treatment, respectively. Moreover, administration of PPP by intraperitoneal injection significantly suppressed the tumor growth of xenografted NPC in nude mice. Taken together, these results suggest targeting IGF-1R by PPP may represent a new strategy for treatment of NPCs with positive IGF-1R expression.

Fibroangiolipoma of palatine tonsil: A case report and literature review.

Lipomas of the palatine tonsil are rare benign neoplasms in clinical practice. We present a case of palatine tonsillar fibroangiolipoma in a 50-year-old Chinese male with a history of multiple lipomas on the back and extremities. It was diagnosed based on histological examination and integrated analysis. Good wound healing and no evidence of recurrence were noted within 6 months follow-up after tonsillectomy. This article also puts a spotlight on the differential diagnosis of benign tonsillar tumors and reviewed recent relevant literature.

MicroRNA-379-5p/YBX1 Axis Regulates Cellular EMT to Suppress Migration and Invasion of Nasopharyngeal Carcinoma Cells.

BACKGROUND: Epithelial-mesenchymal transition (EMT) is a major actor modulating the metastasis of nasopharyngeal carcinoma (NPC). Increasing evidence indicates that microRNAs (miRs) are the important regulators of EMT program. However, the potential roles and underlying mechanisms of miR­379-5p in regulating EMT of NPC cells remain unclear. METHODS: miR-379-5p expression levels in human NPC tissues and cell lines were detected via quantitative real-time PCR (qRT-PCR). Then, the correlations between miR-379-5p expression in NPC tissues and clinicopathologic features and patients' prognosis were analyzed. The effect of miR-379-5p on the expression of EMT markers in NPC cells was evaluated by Western blot and qRT-PCR. NPC cells' migration and invasion were evaluated in vitro by Transwell migration and invasion assays, respectively. The target of miR-379-5p was predicted with three publicly available databases and further validated with dual-luciferase reporter assay, qRT-PCR, and Western blot. RESULTS: The expression of miR-379-5p was significantly decreased in NPC tissues, and its low expression was significantly associated with multiple unfavorable clinicopathological factors and poor prognosis of NPC patients. Meanwhile, miR-379-5p was downregulated in NPC cell lines, and its exotic expression inhibited EMT to reduce the migration and invasion of NPC cells. Furthermore, Y-box binding protein 1 (YBX1) was identified and validated as a direct target of miR-379-5p, and restoring YBX1 expression could reverse the inhibitive effect of miR-379-5p on NPC cell EMT, migration and invasion. CONCLUSION: Taken together, our findings indicate that miR-379-5p inhibits the EMT of NPC cells to reduce their migration and invasion abilities by post-transcriptionally suppressing YBX1 expression, providing a novel potential treatment target for NPC patients.

Long non-coding RNA 319 facilitates nasopharyngeal carcinoma carcinogenesis through regulation of miR-1207-5p/KLF12 axis.

Long non-coding RNAs (lncRNAs) have been wildly verified to modulate multiple tumorigeneses, especially nasopharyngeal carcinoma (NPC). In present study, we aim to investigate the role of LINC00319 in the NPC carcinogenesis. It was indicated that LINC00319 was markedly increased in NPC tissues and cells in comparison to their corresponding controls. Moreover, the aberrant overexpression of LINC00319 indicated the poor prognosis of NPC patients. Silence of LINC00319 was able to suppress NPC cell growth in vitro while overexpression of LINC00319 inversed this process. Moreover, in vivo tumor xenografts were established using CNE-1/SUNE-1 cells to investigate the function of LINC00319 in NSCLC tumorigenesis. Rescue assay was performed to further confirm that LINC00319 contributed to NPC progression by regulating miR-1207-5p/KLF12 signal pathway. Taken together, our study discovered the oncogenic role of LINC00319 in clinical specimens and cellular experiments, showing the potential LINC00319/miR-1207-5p/KLF12 pathway. This results and findings provide a novel insight for NPC tumorigenesis.

Insulin-like growth factor type-I receptor-dependent phosphorylation of extracellular signal-regulated kinase 1/2 but not Akt (protein kinase B) can be induced by picropodophyllin.

The initial event upon binding of insulin-like growth factor 1 to the insulin-like growth factor type-I receptor (IGF-1R) is auto-phosphorylation of tyrosine residues within the activation loop of the kinase domain followed by phosphorylation of other receptor tyrosine residues and the subsequent activation of the intracellular signaling cascades. We found recently that the cyclolignan picropodophyllin (PPP) inhibits phosphorylation of IGF-1R and phosphatidyl-3 kinase/Akt (protein kinase B) signaling molecules without interfering with the highly homologous insulin receptor. Furthermore, PPP causes regression of tumor grafts and substantially prolongs the survival of animals with systemic tumor disease. It is of interest that we show here that short treatments with PPP activate the intracellular extracellular signal-regulated kinase (ERK) signaling. Our data suggest that PPP induces IGF-1R ubiquitination and in turn activates ERK1/2. The PPP-induced ERK activation requires IGF-1R because PPP is not able to induce ERK phosphorylation in IGF-1R-negative cells or in cells in which the receptor is knocked down by small interfering RNA. Moreover, in the absence of Mdm2, an E3 ligase that has been shown previously to be involved in IGF-1R ubiquitination, the phosphorylation of ERK did not occur. Thus, apart from inhibiting the receptor activity, PPP can induce IGF-1R ubiquitination and stimulate ERK in an Mdm2-dependent manner. This response could contribute to the apoptotic effect of PPP.

Expression and growth dependency of the insulin-like growth factor I receptor in craniopharyngioma cells: a novel therapeutic approach.

Craniopharyngioma is a rare benign intracranial epithelial tumor that, however, often recurs and sometimes kills the affected patients, one-third of which are children. In many cases, the patients acquire growth hormone deficiency and postoperatively need substitution. Generally, growth hormone promotes local release of insulin-like growth factor I (IGF-I), which in turn activates the IGF-I receptor (IGF-IR) if present. Together, these circumstances raise the question whether IGF-IR may be involved in craniopharyngioma growth. To address this issue, we analyzed phenotypically well-characterized primary low-passage craniopharyngioma cell lines from nine different patients for IGF-IR expression and IGF-I dependency. Two of the cell lines showed no/very low expression of the receptor and was independent on IGF-I, whereas five cell lines exhibited a strong expression and was clearly contingent on IGF-I. The two remaining cell lines had low receptor expression and IGF-I dependency. Upon treatment with an IGF-IR inhibitor, cells with high IGF-IR expression responded promptly with decreased Akt phosphorylation followed by growth arrest. These responses were not seen in cells with no/very low receptor expression. Growth of cell lines with low IGF-IR expression was only slightly affected by IGF-IR inhibition. Taken together, our data suggest that IGF-IR may be involved in the growth of a subset of craniopharyngiomas and points to the possibility of the involvement of IGF-IR inhibitors as a treatment modality to obtain complete tumor-free conditions before growth hormone substitution.

Long non-coding RNA EWSAT1 promotes human nasopharyngeal carcinoma cell growth in vitro by targeting miR-326/-330-5p.

Long non-coding RNA (lncRNA) Ewing sarcoma associated transcript 1 (EWSAT1) has been identified as an oncogene, and its dysregulation is closed corrected with tumor progression in Ewing sarcoma. Recently, high-through put analysis reveals that EWSAT1 is also highly expressed in human nasopharyngeal carcinoma (NPC). However, whether the aberrant expression of EWSAT1 in NPC is corrected with malignancy or prognosis has not been expounded. Herein, we identified that EWSAT1 was up-regulated in NPC tissues and cell lines, and higher expression of EWSAT1 resulted in a markedly poorer survival time. EWSAT1 over-expression facilitated, while EWSAT1 silencing impaired cell growth in NPC. In addition, mechanistic analysis demonstrated that EWSAT1 up-regulated the expression of miR-326/330-5p clusters targeted gene cyclin D1 through acting as a competitive 'sponge' of miR-326/330-5p clusters. Collectively, our data revealed that EWSAT1 promotes NPC cell growth in vitro through up-regulating cyclin D1 partially via 'spongeing' miR-326/330-5p clusters.

Effects of TESTIN gene expression on proliferation and migration of the 5-8F nasopharyngeal carcinoma cell line.

PURPOSE: To investigate effects of the TESTIN (TES) gene on proliferation and migration of highly metastatic nasopharyngeal carcinoma cell line 5-8F and the related mechanisms. MATERIALS AND METHODS: The target gene of human nasopharyngeal carcinoma cell line 5-8F was amplified by PCR and cloned into the empty plasmid pEGFP-N1 to construct a eukaryotic expression vector pEGFP-N1-TES. This was then transfected into 5-8F cells. MTT assays, flow cytometry and scratch wound tests were used to detect the proliferation and migration of transfected 5-8F cells. RESULTS: A cell model with stable and high expression of TES gene was successfully established. MTT assays showed that the OD value of 5-8F/TES cells was markedly lower than that of 5-8F/GFP cells and 5-8F cells (p<0.05). Flow cytometry showed that the apoptosis rate of 5-8F/TES cells was prominently increased compared with 5-8F/GFP cells and 5-8F cells (p<0.05). In vitro scratch wound assays showed that, the width of the wound area of 5-8F/TES cells narrowed slightly, while the width of the wound area of 5-8F/ GFP cells and 5-8F cells narrowed sharply, suggesting that the TES overexpression could inhibit the migration ability. CONCLUSIONS: TES gene expression remarkably inhibits the proliferation of human nasopharyngeal carcinoma cell line 5-8F and reduces its migration in vitro. Thus, it may be a potential tumor suppressor gene for nasopharyngeal carcinoma.

Targeting the insulin-like growth factor-1 receptor by picropodophyllin as a treatment option for glioblastoma.

Glioblastoma (GB) is the most common malignant brain tumor in adults. It has limited treatment opportunities and is almost exclusively fatal. Owing to the central role the insulin-like growth factor-1 receptor (IGF-1R) plays in malignant cells, it has been suggested as a target for anticancer therapy including GB. The cyclolignan picropodophyllin (PPP) inhibits IGF-1R without affecting the highly homologous insulin receptor. Here, we show that PPP inhibits growth of human GB cell lines along with reduced phosphorylation of IGF-1R and AKT. In vivo, PPP-treatment causes dramatic tumor regression not only in subcutaneous xenografts but also in intracerebral xenografts, indicating passage of PPP across the blood-brain barrier.

[Expression and clinical significance of insulin like growth factor-1 protein in nasopharyngeal carcinoma].

OBJECTIVE: To evaluate the expression of IGF-1R protein and its clinical signification in nasopharyngeal carcinoma. METHOD: SP immunohistochemical staining was performed to detect the expression of IGF-1R in 63 cases of NPC and 21 inflammatory nasopharyngeal mucosa; and the expression of IGF-1R in three NPC cell lines and chronic nasopharyngitis tissues was examined by western blots. RESULT: The positive rates of IGF-1R were 82.5% and 14.3% respectively in 63 cases of NPC and 21 inflammatory nasopharyngeal mucosa; it also showed stronger positive expression in Stage III-IV of NPC than that in stage I-II (P < 0 01); IGF-1R positive expression in cases of NPC with cervical lymph node metastasis was significantly higher than that of NPC without lymph node metastasis (P < 0.05); it more often appears in recurrent cases than in primary cases (P < 0.05); IGF-1R positive expression had a significantly stronger in patients whose survival time less than 5 years than that in patients whose survival time over 5 years (P < 0.01). CONCLUSION: Over-expression of IGF-1R is related to poor differentiation of NPC, also may be linked with oncogenesis and development of NPC. The over-expression of IGF-1R may be one of a prognostic factors in NPC. The IGF-1R might be a potential therapeutic target in nasopharyngeal cancer.

IGF-1R tyrosine kinase expression and dependency in clones of IGF-1R knockout cells (R-).

Insulin-like growth factor 1 receptor (IGF-1R) plays many crucial roles in cancer, like anti-apoptotic activity and necessity for transformation. IGF-1R knockout cells (R-) represent a useful tool for molecular mapping of biological properties of the receptor. R- cells have been shown to be refractory to transformation by viral and cellular oncogenes, highlighting the necessity of this receptor for transformation. Surprisingly, more recent studies have shown that these cells can undergo spontaneous transformation. This observation raises the question as whether R- cells over the years have acquired some properties mimicking those of IGF-1R. Using an IGF-1R inhibitor (cyclolignan PPP) we have identified clones of R- (R-s) that are sensitive to this compound. Since, PPP is closely related to podophyllotoxin, which is an efficient microtubule inhibitor, we first investigated if such a mechanism could explain the sensitivity to PPP. However, highly purified PPP showed no or very slight tubulin binding. Further analysis of R-s revealed expression of a 90 kDa protein being reactive to IGF-1R beta-subunit antibodies. This protein was weakly but constitutively tyrosine phosphorylated and was downregulated by siRNA targeting IGF-1R. This downregulation was paralleled by decreased R-s survival. Taken together, our study suggests that clones of R- express IGF-1R activity and dependency, which in turn may explain that R- can undergo spontaneous transformation.

[Expression of BAG-1 and p53 in laryngeal squamous cell carcinoma and their clinical significance].

OBJECTIVE: To study the expression of BAG-1 and p53 in laryngeal squamous cell carcinoma (LSCC), and the relation between BAG-1 and p53. METHOD: BAG-1 and p53 protein were examined by using the method of immunohistochemical SP staining method in 40 cases of LSCC, 27 cases of pap- a-cancerous tissues and 20 cases of polyp specimens from vocal cord tissues. RESULT: The positive expression of BAG-1 in. LSCC was significantly higher than that in para-cancerous tissues and polyp specimens from vocal cord tissues (P < 0.05). The positive expressions of BAG-1 were related to the prognosis (P < 0.05), and BAG-1 in LSCC with nucleus staining had worse prognosis than that with cytoplasm staining (P < 0.05). The positive expression of p53 in LSCC was significantly higher than that in para-cancerous tissues and polyp specimens from vocal cord tissues (P < 0.05). The positive expressions of p53 were related to the differentiation grades, cervical lymph node metastasis and prognosis (P < 0.05). There was no significance correlation between BAG-1 and p53 expression (P > 0.05). CONCLUSION: Over expression of BAG-1 and p53 play an important role in the process of development of LSCC; BAG-1 may be a biological predictor of early period in LSCC, especially nuclear BAG-1 localization may be a prediction of unfavorable outcome. Expression of p53 is related with cervical lymph node metastasis, and that is a prognostic target.