A Pilot Study on Immunohistochemical Expressions of NF-ĸB, Cyclin-D1, VEGF, and Cox-2 in Advanced Stage Laryngeal Carcinoma

BACKGROUND: Progression of laryngeal carcinoma can be classified with the clinical staging, however there are different patterns of progressions observed in the patient with the same clinical stage which also affects their prognoses. Therefore biomarkers should be used. Nuclear factor (NF)-ĸB, Cyclin-D1, vascular endothelial growth factor (VEGF), cyclooxygenase (Cox)-2 have been reported for laryngeal carcinoma. However, it is still unclear how these markers are expressed and correlated in advanced stage laryngeal carcinoma. Therefore current study was conducted to investigate the expressions of NF-ĸB, Cyclin-D1, VEGF and Cox-2 and their correlations in advanced stage laryngeal carcinoma.METHODS: Subjects were recruited and laryngeal biopsies were collected, fixed in formalin and prepared for immunohistochemistry. The immunohistochemistry was performed using mouse monoclonal anti-NF-kB p65, anti-Cyclin-D12 anti-VEGF, and anti-Cox-2 antibodies. The immunohistochemistry results were documented and measured using ImmunoRatio. Pearson or Spearman correlation test was used based on the results of Shapiro-Wilk test of normality. A p-value of less than 0.05 is considered statistically significant.RESULTS: Twelve male subjects were included in this study. Expressions of NF-ĸB, Cyclin-D1, VEGF dan Cox-2 were clearly observed. Mean of NF-ĸB, Cyclin-D1, VEGF dan Cox-2 IHC expression levels measured with ImmunoRatio were 57.50±20.06%, 45.00±24.31%, 43.33±17.23% and 40.42±16.98%, respectively. There was significant correlation between the expressions of VEGF dan Cox-2 (p=0.031, r=0.622).CONCLUSION: Since correlation between the VEGF and Cox-2 expressions was statistically significant, VEGF and Cox-2 might have important roles in the growth, invasion and metastasis of laryngeal carcinoma.KEYWORDS: advanced stage laryngeal carcinoma, immunohistochemistry, NF-ĸB, Cyclin-D1, VEGF, Cox-2

Ứng dụng nội soi dải ánh sáng hẹp kết hợp nội soi vi phẫu bằng laser CO2 trong chẩn đoán và điều trị ung thư hạ họng, thanh quản giai đoạn sớm

TÓM TẮT Mục tiêu: Đánh giá kết quả ứng dụng nội soi dải ánh sáng hẹp kết hợp nội soi vi phẫu bằng Laser CO2 trong chẩn đoán và điều trị ung thư hạ họng và ung thư thanh quản giai đoạn sớm. Phương pháp nghiên cứu: Nghiên cứu tiến cứu, có can thiệp lâm sàng trên 18 bệnh nhân ung thư hạ họng, ung thư thanh quản ở giai đoạn sớm, được phát hiện tổn thương ác tính bằng nội soi dải ánh sáng hẹp, điều trị phẫu thuật nội soi vi phẫu bằng Laser CO2. Kết quả: Tuổi trung bình mắc bệnh là 63,8 ± 12,6; bệnh nhân có hút thuốc lá chiếm 61,1%; triệu chứng khàn tiếng chiếm 72%, ung thư giai đoạn T1a có tỷ lệ 72,2%; IPCL type V-1 là 72,2%; khả năng kiểm soát bờ rìa là 83,3%, không có tai biến trong phẫu thuật 17/18 trường hợp và di chứng sau phẫu thuật tháng thứ 1 là tăng sinh mô hạt (77,8%), tháng thứ 3 là dính mép trước dây thanh (27,8%). Kết luận: Nội soi dải ánh sáng hẹp có giá trị phát hiện sớm ung thư hạ họng, ung thư thanh quản. Phẫu thuật nội soi vi phẫu bằng Laser CO2 ung thư hạ họng, ung thư thanh quản ở giai đoạn sớm mang lại kết quả tốt, tỷ lệ tai biến và biến chứng sau phẫu thuật thấp, bảo tồn được tối đa chức năng phát âm, hô hấp và nuốt. ABSTRACT UTILITY OF NARROW - BAND IMAGING ENDOSCOPY COMBINED WITH CO2 LASER ENDOSCOPIC MICROSURGERY IN DIAGNOSIS AND TREATMENT OF EARLYSTAGE HYPOPHARYNGEAL AND LARYNGEAL CARCINOMA Objective: To evaluate the results of applying narrow band imaging endoscopy combined with CO2 laser endoscopic microsurgery to diagnose and treat early stage hypopharyngeal and laryngeal carcinoma. Methods: A prospective study was conducted on 18 patients with early stage hypopharyngeal and laryngeal carcinoma. Results: The mean age was 63.8 ± 12.6; smoker was 61.1%, the rate of hoarseness was 72%, T1a tumor stage was 72.2%, IPCL type V-1 was 72,2%, marginal tumor control was 83.3%. There were of 17/18 case with no complication during operation. The rate of extensive granuloma was 77.8% in the first month, and of anterior commissure adhesion was 27.8% in the third month post - surgery. Conclusion: Narrow band imaging endoscopy was a valuable tool in screening for early stage hypopharyngeal and laryngeal carcinoma. CO2 laser endoscopic microsurgery was an effective method. The rate of complication during and after surgery was low. This method restores the maximum function of speech, breathing and swallowing. Keywords: Early - stage pharyngeal cancer, early - stage laryngeal cancer, narrow - band imaging endoscopy, CO2 laser endoscopic microsurgery.

Effect of Notch1 Signaling on Cellular Proliferation and Apoptosis of Human Laryngeal Carcinoma

The pathological processes of occurrence and development of malignancies include the excessive proliferation and apoptosis resistance of neoplastic cells. The study aims to identify the effects of Notch1 signaling on the proliferation and apoptosis of laryngeal cancer cells in hypoxic microenvironment. Notch1 and Ki-67 expression in laryngeal squamous cell carcinoma (LSCC) tissue samples were detected by immunohistochemistry. The apoptotic index (AI) of LSCC was evaluated by TUNEL method. In laryngeal cancer cells, small interfering RNA (siRNA) technology was to inhibit Notch1 expression. Meanwhile, Real-time PCR detected Notch1, Hes1 and Hey1 mRNA expression, and Western blot detected Notch1 and Notch1 intracellular domain (N1ICD) protein expression. Annexin V-FITC/propidium iodide staining and Cell Counting Kit‑8 methods measured cell apoptosis and proliferation, respectively. Notch1 expression was detected in 63.55％(68/107) of LSCC samples and was significantly related to the proliferation index (PI) (P < 0.05) and AI (P < 0.05) in LSCC tissues. Furthermore, it was confirmed that hypoxia could induce proliferation and inhibit apoptosis of laryngeal carcinoma cells (P < 0.05). Meanwhile, Notch1 expression and Notch1 signaling activity could be upregulated by hypoxia (P < 0.05). In contrast, suppression of Notch1 signaling activity in hypoxic neoplastic cells could obviously decrease cell proliferation and increase cell apoptosis (both P<0.05). Our study has demonstrated that hypoxia may promote cell proliferation and inhibit cell apoptosis of laryngeal carcinoma. Notch1 signalling may exert a pivotal role in regulating the proliferation and apoptosis resistance of laryngeal cancer cells under hypoxia.

miR-206 Inhibits Laryngeal Carcinoma Cell Multiplication, Migration, and Invasion

Laryngeal carcinoma (LC) is one of the common human cancer types. MicroRNAs (miRNAs) were reported to be the essential regulators in cancer diagnosis, treatment, and prognosis. It was reported that miR-206 expression was reduced in various neoplastic diseases. However, the role and functional mechanism of miR-206 in LC progression remain unclear. In this research, miR-206 was found to be associated with tumor-node-metastasis (TNM) staging. In addition, the area under the curve (AUC) of miR-206 was 0.902 for diagnosis of LC and 0.854 for differential diagnosis of stage I-II and stage III-IV patients. Low expression of miR-206 was associated with poor prognosis of LC patients. miR-206 expression was an independent factor affecting the prognosis of LC patients, as revealed by the Cox regression analysis. In vitro experiments demonstrated that miR-206 overexpression reduced cell multiplication, invasion, and migration and increased cell apoptosis in LC cells. Moreover, SOX9 was a target of miR-206, and miR-206 negatively regulated SOX9 expression. Collectively, miR-206 might be a promising biomarker with diagnostic and prognostic value for LC, and the miR-206/SOX9 axis might be a candidate target for LC therapy.

Undifferentiated laryngeal carcinoma with hyaline bodies in a cat

Background Primary laryngeal neoplasms are rare in cats, with lymphoma and squamous cell carcinoma being the most commonly diagnosed tumour types. These tumours are usually highly aggressive, difficult to treat, and have a poor prognosis. Here an undifferentiated laryngeal carcinoma with hyaline bodies in a cat is reported. Case presentation A 13-year-old cat was presented for progressive respiratory signs. Diagnostic procedures revealed a partially obstructive laryngeal mass. Cytology was compatible with a poorly differentiated malignant tumour, with neoplastic cells frequently containing large intracytoplasmic hyaline bodies. After 1 month the patient was euthanised due to a worsening clinical condition and submitted for post-mortem examination, which confirmed the presence of two laryngeal masses. Histopathology confirmed the presence of an undifferentiated neoplasm with marked features of malignancy. Strong immunolabelling for pancytokeratin led to a diagnosis of undifferentiated carcinoma, however, histochemical and immunohistochemical investigations could not elucidate the origin of the large intracytoplasmic hyaline bodies observed in tumour cells, which appeared as non-membrane bound deposits of electron-dense material on transmission electron microscopy. Conclusion This is the first report of primary undifferentiated laryngeal carcinoma in a cat. Our case confirms the clinical features and the short survival that have been reported in other studies describing feline laryngeal tumours. Moreover, for the first time in feline literature, we describe the presence of intracytoplasmic hyaline bodies in neoplastic cells that were compatible with the so-called hyaline granules reported in different human cancers and also in the dog.

Role and mechanism of Glut-1 and H+/K+-ATPase expression in pepsin-induced development of vocal cord leukoplakia

Purpose We investigated the role of Glut-1 and H+/K+-ATPase expression in pepsin-induced development of human vocal cord leukoplakia cells (HVCLCs). Next, we analyzed the relationship between Glut-1 and H+/K+-ATPase expression with the clinicopathological features of laryngeal carcinoma. Methods Glut-1 and H+/K+-ATPase expression levels in HVCLCs were determined after treatment with artificial gastric juice containing pepsin and laryngeal carcinoma tissues. Results Exposure to pepsin-containing artificial gastric juice significantly enhanced the migration and proliferation of VSCLCs in a time-dependent manner. The apoptotic rate of VSCLCs decreased over time after exposure to pepsin and reached a nadir on day 7 (p < 0.01). With increasing duration of exposure to pepsin, the proportion of VSCLCs in G0/G1 phase decreased and the proportions in the S and G2/M phases significantly increased (p < 0.05). After treatment with pepsin-containing artificial gastric juice, RT-PCR and Western blotting showed that the expression of Glut-1 and H+/K+-ATPase α, β significantly increased in HVCLCs compared to in the absence of pepsin (p < 0.05). The expression of Glut-1 and H+/K+-ATPase α, β gradually increased from vocal cord leukoplakia (VLC) to laryngeal carcinoma (p < 0.05). Lentivirus-mediated inhibition of Glut-1 expression in VCL significantly inhibited the cells’ migration and proliferation (p < 0.05) but enhanced their apoptosis (p < 0.05). Also, inhibition of Glut-1 expression resulted in an increased proportion of cells in G0/G1 phase and a significantly decreased proportion in G2/M phase (p < 0.05). Conclusions Elevated Glut-1 expression may promote the development of VCL by upregulating laryngeal H+/K+-ATPase expression to reactivate absorbed pepsin, thus damaging the laryngeal mucosa.

Epstein-Barr Virus-Associated Carcinoma of the Larynx: A Systematic Review with Meta-Analysis

Over the past few decades, several publications have investigated the role of Epstein-Barr virus (EBV) in head and neck squamous cell carcinomas, and an increasing number of them have shown its presence in laryngeal tumors. The purpose of this meta-analysis was to evaluate the association of EBV with laryngeal carcinoma. The search was carried out in two databases, Scopus and PubMed, using the following terms: “Epstein-Barr virus” and “laryngeal carcinoma”. A total of 187 records were found, of which 31 were selected for meeting the inclusion and exclusion criteria. The meta-analysis yielded an overall pooled prevalence of 43.72% (95% confidence interval (CI): 34.35–53.08). Studies carried out in Europe and Eurasia had slightly higher pooled prevalence than other subgroups, while the prevalence of studies performed in developed countries was higher than in developing countries (46.37% vs. 34.02%). Furthermore, laryngeal carcinoma occurred almost three times as often among EBV-infected individuals compared to those without EBV infection (odds ratio = 2.86 (95% CI: 1.18–6.90); Begg’s test, p = 0.843 and Egger’s test, p = 0.866). Our findings support the idea that EBV is related to laryngeal carcinoma. However, further studies are needed before recognizing a definitive etiological role of EBV in the development and/or progression of laryngeal carcinomas.