Elevation of miR-125b-5p is related to improved prognosis in laryngeal squamous cell carcinoma and inhibits the malignancy and glycometabolic disorder by targeting MAP3K9.

Laryngeal squamous cell carcinoma (LSCC) is the most common malignant tumor in the head and neck cancer, with a poor prognosis. As we know, microRNAs (miRNAs) play a vital role in the initiation and development of various cancers including LSCC. In this study, we explored the role of miR-125b-5p and its downstream regulatory pathway in LSCC. Our data demonstrated that miR-125b-5p expression was significantly downregulated in LSCC tissues and cells. LSCC patients with high expression of miR-125b-5p had higher overall survival (OS) and were closely related to the clinical stage. Overexpression of miR-125b-5p impaired viability and glycolysis, and facilitated apoptosis in LSCC cells. And miR-125b-5p silencing had the opposite effects. Bioinformatics website predicted that MAP3K9 was one of the potential target genes of miR-125b-5p. Cell experiments demonstrated that miR-125b-5p repressed the MAP3K9 levels by directly targeting MAP3K9. Additionally, the negative correlation between miR-125b-5p and MAP3K9 was validated in LSCC tissues. Overexpression of MAP3K9 attenuated the inhibitory effect of miR-125b-5p on viability and glycolysis, and the pro-apoptosis effect of miR-125b-5p in LSCC cells. Furthermore, in vivo experiments demonstrated that tumor growth was hampered in AMC-HN-8 cells transfected with miR-125b-5p mimic. In contrast, the knockdown of miR-125b-5p reduced tumor growth in vivo. Meanwhile, the in vivo immunohistochemistry and TUNEL assays suggested that the miR-125b-5p overexpression restrained cell proliferation and promoted apoptosis via targeting MAP3K9. Overall, these above results suggested that miR-125b-5p suppressed proliferation and glycolysis, and promoted apoptosis by directly targeting MAP3K9 in LSCC cells. Thus, miR-125b-5p acts as a tumor suppressor miRNA and the miR-125b-5p/MAP3K9 axis may be a promising candidate for LSCC treatment.

The value of circulating CYFRA21-1 expression in patients with nasopharyngeal carcinoma: a study of 529 subjects.

BACKGROUND: Early diagnosis of nasopharyngeal carcinoma (NPC) needs more reliable biomarkers. The aim of this study was to investigate serum cytokeratin 19 fragment 21.1 (CYFRA21-1) as an NPC biomarker based on data from a large sample. METHODS: From October 2010 to February 2014, 529 subjects were enrolled and divided into three groups-NPC group (n = 274), healthy control group (n = 175) and nasal inflammatory disease group (n = 80). Serum CYFRA21-1 levels were measured prior to radiotherapy/chemoradiotherapy, and their associations with T, N, and clinical classification were determined. Receiver operating characteristic curve analysis was performed to discriminate the NPC group from the healthy control and nasal inflammatory disease groups. Three Epstein-Barr virus (EBV) antibodies and their correlations with serum CYFRA21-1 levels were analyzed. RESULTS: Pretreatment serum CYFRA21-1 levels were significantly elevated in the NPC group compared with the other groups (p < 0.01), Furthermore, serum CYFRA21-1 levels decreased significantly after radiotherapy (p < 0.01). Serum CYFRA21-1 levels were closely related to T, N, and clinical classifications. The area under the curve, sensitivity and specificity of the serum CYFRA21-1 levels in the NPC patients were 0.89, 0.87 and 0.83, respectively. Strong correlations were observed between serum CYFRA21-1 levels and EBV antibodies. CONCLUSION: Serum CYFRA21-1 may be a reliable and effective biomarker for NPC.

Assessment of the effect of local application of amifostine on acute radiation-induced oral mucositis in guinea pigs.

The aim of present study was to assess the radioprotective effects of the local application of amifostine to treat acute buccal mucositis in guinea pigs. A total of 32 guinea pigs were randomized into four groups: (Group A) topically administered 50 mg of amifostine plus radiotherapy (RT); (Group B) 100 mg amifostine plus RT; (Group C) normal saline plus RT; and (Group D) normal saline plus sham RT. The opportunity for administration was 15 min before irradiation. When administered, the cotton pieces that had been soaked with 0.5 ml amifostine solution or saline were applied gently on the buccal mucosa of each guinea pig for 30 min. The animals in Groups A, B and C were irradiated individually with a single dose of 30 Gy to the bilateral buccal mucosa. Eight days after irradiation, the animals were scored macroscopically; they were then euthanized, and the buccal mucosal tissues were processed for hematoxylin-eosin staining and ICAM-1 immunohistochemical analysis. In Groups A and B, the mean macroscopic scores were 2.9 ± 0.6 and 2.4 ± 1.1, respectively. There was no significant difference between the two groups (P > 0.05). However, when they were separately compared with Group C (4.4 ± 0.7), a noticeable difference was obtained (P < 0.05). No mucositis was observed in Group D. Comparisons of the expression of ICAM-1 were in agreement with the macroscopic data. Histologically, superficial erosion, exudate and ulcer formation were all observed in the RT groups; only the severity and extent were different. The microscopic observations in the amifostine-treated groups were better than in Group C. The results demonstrated that topical administration of amifostine to the oral mucosa is effective treatment of acute radiation-induced mucositis.

Frequent epigenetic inactivation of Myocardin in human nasopharyngeal carcinoma.

BACKGROUND: Epigenetic silencing of tumor suppressor genes plays an important role in nasopharyngeal carcinoma (NPC) tumorigenesis. In the present study, we explore a novel target gene of epigenetic silencing in NPC, Myocardin, which is inactivated by promoter hypermethylation. METHODS: Transcriptional expression levels of Myocardin were evaluated by reverse transcription-polymerase chain reaction (RT-PCR). Methylation status was addressed by methylation-specific PCR and bisulfite genomic sequencing. RESULTS: Myocardin mRNA expression was inactivated in 4 of 5 NPC cell lines. Myocardin was aberrantly methylated in 4 of 5 NPC cell lines (80%) and in 48 of 65 NPC primary tumors (73.8%, but not in any of the 12 normal nasopharyngeal tissues tested. Myocardin expression could be reactivated in NPC cells after treatment with the demethylating agent 5-aza-2'-deoxycytidine (5-aza-dC). CONCLUSIONS: Epigenetic inactivation of Myocardin is a frequent and tumor-specific event in NPC. Our findings suggest that Myocardin is a candidate tumor suppressor gene in NPC.

A study of middle ear function in the treatment of nasopharyngeal carcinoma with IMRT technique.

PURPOSE: This study evaluates the difference in damage to middle ear function with CRT and IMRT techniques in the treatment of nasopharyngeal carcinoma (NPC). We explore the isthmus of the Eustachian tube (ET) as the key anatomic site for the prevention of radiation-induced otitis media with effusion. METHODS AND MATERIALS: Eighty-two patients with NPC were divided into two groups: 40 patients treated with CRT and 42 patients treated with IMRT. The difference between dosage over the middle ear cavity and the isthmus of the ET was evaluated in both CRT group and IMRT group. All patients underwent hearing tests including pure tone audiometry and impedance audiometry before and after RT. RESULTS: The dosage difference to the middle ear cavity and isthmus between these two groups was statistically significant (p<0.05). The difference in hearing test results between these two groups was also statistically significant (p<0.05). If we limited the dose to the middle ear cavity under 34 Gy and the dose to the isthmus under 53 Gy with IMRT, we may decrease radiation-induced OME even with the larger 2.25 Gy fraction size. CONCLUSIONS: IMRT may have better protected the middle ear function compared with the CRT technique, even with larger fraction sizes than for the conventional CRT technique.

Analysis of anatomical factors controlling the morbidity of radiation-induced otitis media with effusion.

PURPOSE: To analyze the anatomical factors controlling the morbidity of radiation-induced otitis media with effusion (OME) and determine how to best preserve middle ear function when treating nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: Forty patients with nasopharyngeal carcinoma undergoing 3-D radiotherapy (RT) planning and curative RT were analyzed retrospectively. The difference in dosage over the middle ear cavity and the isthmus of the Eustachian tube (ET) was evaluated. Pure tone audiometry and impedance audiometry tests were performed before and after RT. RESULTS: Mean dosages over the isthmus of the ET for acoustic impedance and pure tone audiometry were recorded. Differences in dosage among the three classifications of unchanged, improved, and worsened ears were statistically significant. CONCLUSION: There was a correlation between the morbidity of radiation-induced OME and the radiation dosage over the middle ear cavities. Decreased OME morbidity was observed when the dosage over the isthmus of the ET was below 52 Gy and the dosage over middle ear cavity was below 46 Gy.

The relationship between the protective effect of amifostine and decreased intercellular adhesion molecule 1 expression.

PURPOSE: The objective of our study was to characterize the relationship between the protective effect of amifostine and decreased intercellular adhesion molecule 1 (ICAM-1) expression in early-phase, radiation-induced otitis media and to illustrate the possible mechanism of early-phase radiation-induced otitis media. MATERIALS AND METHODS: A comparison study of middle ear tissue was performed by the expression of ICAM-1 and the electron microscope from total 38 guinea pigs. Group A, consisting of 2 pigs, was used as control, and these pigs were not irradiated. Groups B, C, D, and E, consisting of 9 pigs each, were irradiated. Sterile saline was administered intraperitoneally to the pigs in groups B and D before irradiation, and amifostine was administered intraperitoneally as an aqueous solution 30 minutes before irradiation to the pigs in groups C and E. The pigs in groups B and C were killed on the second day after irradiation, and the pigs in groups D and E were killed 30 days after irradiation. RESULTS: Intercellular adhesion molecule 1 was strongly expressed in the middle ear mucosa of the irradiated pigs after a 45-Gy dose of radiation was administered. Enhanced ICAM-1 expression was accompanied by pathomorphologic changes in the middle ear tissue. Scanning electron microscopy demonstrated the changes. Intercellular adhesion molecule 1 expression in the mucosa of the groups killed on the second day was stronger than that in the mucosa of the groups killed 30 days after irradiation. Amifostine protected the middle ear from radiation injury, and we found that the expression of ICAM-1 in the middle ear mucosa was down-regulated. However, slight expression of ICAM-1 remained 30 days after irradiation. CONCLUSIONS: Irradiation increased the expression of ICAM-1 in the middle ear mucosa. Amifostine protected the middle ear from early irradiation injury. There was a relationship between oxygen free radicals derived from irradiation and up-regulation of ICAM-1 expression. Continuous ICAM-1 expression might be related to stenosis of the eustachian tube.