A pilot study reveals the potential of miR-31-3p and miR-196a-5p as non-invasive biomarkers in advanced laryngeal cancer

Introduction: Recently, miRNAs have become popular molecules used as non-invasive biomarkers in cancer diseases. Aim: The aim of the study was to explore the expression of four miRNAs isoforms: miR-31-3p, miR-196a-5p, miR-210-3p and miR-424-5p in plasma and tissue samples from patients with advanced laryngeal squamous cell carcinoma (LSCC) and healthy controls. Materials and methods: Fresh-frozen tumour and normal laryngeal tissue as well as plasma samples were obtained from 22 patients diagnosed with advanced LSCC. The control group included plasma samples from 21 cancer-free volunteers. Total RNA (including miRNAs) extraction, reverse transcription and real time qPCR were the laboratory techniques used in the study. The obtained results were analyzed using SPSS software v. 23. Results: We found that miR-31-3p, miR-196a-5p, and miR-210-3p levels were significantly elevated in laryngeal tumour tissue, but only the levels of miR-31-3p and miR-196a-5p were significantly upregulated in the plasma LSCC target group. Positive correlation was obtained for miR-31-3p (rs=0.443, p=0.039) and miR-196a-5p (rs=0.548; p=0.008) between plasma and adjacent tumour tissue LSCC samples. ROC analyses were used to evaluate the discriminative power of both miRNAs alone and in combination. The combination of miR-31-3p and miR-196a-5p showed best results with AUC=0.978 (95% CI: 0.945&ndash;1.000, p<0.001) with 100% sensitivity and 81% specificity at cut-off: RQ=2.99. Conclusions: Based on this miR-31-3p and miR-196a-5p are proposed as potential biomarkers for validation in larger LSCC group and could be included in a non-invasive miRNAs set for detection of advanced LSCC.

Erchen Plus Huiyanzhuyu Decoction Inhibits the Growth of Laryngeal Carcinoma in a Mouse Model of Phlegm-Coagulation-Blood-Stasis Syndrome via the STAT3/Cyclin D1 Pathway

Erchen plus Huiyanzhuyu decoction (EHD), a Chinese herbal medicine (CHM) formula that consists of Erchen decoction and Huiyanzhuyu decoction, has achieved satisfactory results in the clinic. The main function of EHD is to remove phlegm and blood stasis, and EHD is suitable for phlegm-coagulation-blood-stasis (PCBS) syndrome in laryngeal cancer (LC). In this study, a xenograft mouse model of LC with PCBS syndrome was constructed by feeding a high-fat diet, swimming in ice water, and subcutaneously injecting epinephrine hydrochloride for 2 weeks. Based on the successful Chinese medicine syndrome model, Hep2-luciferase-GFP cells were injected subcutaneously under the armpit of the right upper limb in mice to form tumours. A mouse model of LC with PCBS syndrome was established via heterotopic transplantation. Then, the mice received intragastric administration of different concentrations of EHD daily, and cisplatin (DDP) was intraperitoneally injected every week for 21 days. Tumour fluorescence in mice was measured with a living animal imager on days 7, 14, 21, and 28 during treatment. The results of this experiment confirmed that a mouse model of Chinese medicine syndrome was successfully constructed. Moreover, EHD slowed the growth of xenograft tumours in nude mice; decreased the expression levels of STAT3, p-STAT3, and cyclin D1; and upregulated the expression level of P27. In brief, EHD inhibited laryngeal tumour growth in a xenograft mouse model of PCBS syndrome and regulated the STAT3/cyclin D1 signalling pathway. This study was the first to construct a Chinese medicine xenograft mouse model of LC with PCBS syndrome; in addition, this study clarified that EHD regulated the STAT3/cyclin D1 signalling pathway to inhibit the growth of LC and that EHD may be a promising novel therapeutic compound for the treatment of patients with LC.

Lateral Soft Tissue Neck X-ray a Tool for the Otorhinolaryngology Surgeon

Background: Management of aerodigestive emergencies can prove a challenge especially in this sub-Saharan region where there is a dearth of specialized equipment. In some areas, a plain radiograph may be the only investigative tool readily available to the surgeon. It is routinely done in most of these cases. In our environment health care is often financed by the patients through out of pocket expenses since most do not have health insurance. Affordability of a particular treatment becomes paramount. This study, therefore, is aimed at assessing the use of lateral soft tissue neck X-ray and its reliability as a diagnostic tool in aerodigestive emergencies. Objective: To determine the relevance or effectiveness of plain radiograph of the lateral soft tissue neck in patients with upper aerodigestive emergencies and therefore its use as a tool to the otorhinolaryngology (ORL) surgeon as the first-line investigation in these patients. Patients and Methods: It is a prospective study of all patients that presented to the ENT surgery department of UPTH from April 2018 to April 2019 with upper aerodigestive emergencies. Patients who presented in the ENT clinic, accident and emergency department, children`s emergency wards and the clinics with upper aerodigestive emergencies were recruited in this study. All the patients had X-ray lateral soft tissue of the neck done on presentation and the results were then compared with the final operative findings in the operation theatre. Some patients had also chest radiograph done however; the diagnosis was done based on the lateral soft tissue neck finding. Patients that had radiologic investigation other than radiograph of the lateral soft tissue neck as the main investigative tool were excluded from the study. The radiologist opinion was obtained for all the radiographs as a routine. The results were analyzed using the SPSS 20 and presented in simple statistical tables. Results: Sixteen patients were studied. Males were 14(87.5%) while females were 2(12.5%). Age range was from 1.5 to 77 years with children comprising only 31.25% of the study population. The commonest presenting complaint was voice change 68.75%. Positive findings on the x-ray neck were seen as soft signs in 62.5% and radio-opacity in 18.75%. In n=14(87.5%) the lateral neck x-ray positive findings were corroborated by the operative findings. The tool has a specificity of 100% for both foreign body inhalation/ingestion and laryngeal tumour but the sensitivity of 80% in laryngeal tumours but 100% for the foreign body. Conclusion: Study concludes that lateral soft tissue neck x-ray was found to be a good and useful tool in the diagnosis of a good number of upper aerodigestive emergencies such as foreign body ingestion/inhalation and patients with upper airway obstruction caused by laryngeal tumours and some infective conditions such as a retropharyngeal abscess.

Diagnosis dan Penatalaksanaan Tumor Ganas Laring

Abstrak Laring berperan dalam koordinasi fungsi saluran aerodigestif atas seperti bernafas, berbicara dan menelan.Laring terbagi tiga yaitu supraglotis, glotis dan subglotis. Laring merupakan daerah tersering kedua untuk kasuskarsinoma sel skuamosa kepala-leher, biasanya berhubungan dengan tembakau dan alkohol. Lebih dari 95% kasustumor ganas laring adalah karsinoma sel skuamosa. Pasien tumor ganas laring datang dengan berbagai keluhanseperti disfonia, obstruksi jalan napas, disfagia, odinofagi dan hemoptisis. Diagnosis tumor ganas laring ditegakkanberdasarkan anamnesis, pemeriksaan klinis menggunakan endoskopi kaku, serat optik dan biopsi. Penatalaksanaantumor ganas laring tergantung stadium dengan modalitas berupa operasi, kemoterapi, radiasi atau terapi kombinasi.Dilaporkan kasus laki-laki 53 tahun dengan karsinoma glotis stadium III (T3N0M0) squamous cell ca keratinized welldifferentiated. Penatalaksanaan pada pasien ini dengan melakukan laringektomi total.Kata kunci: Tumor ganas laring, karsinoma, laringektomi, tembakau Abstract Larynx plays a certain role in coordinating functions of the upper aerodigestive tract, such as respiration,speech, and swallowing. The larynx is divided into three region; supraglottic, glottic, and subglottic. Larynx is thesecond most common site for squamous cell carcinoma in the head and neck and usually related to tobacco andalcohol exposure. Primary malignant tumors of the larynx are squamous cell carcinomas can found more than 95% ofcases. Patients with laryngeal tumors usually present with complaints of hoarseness, respiratory obstruction,dysphagia, odynophagia and hemoptysis. Diagnosis of laryngeal cancer is made by medical history, clinicalexamination using a rigid or fiberoptic endoscope and biopsy. Management of laryngeal tumour depends on stadiumwith various modality included surgery, chemotheraphy, radiotheraphy or combined therapy. Reported case of 53years old male with Glottic carcinoma of the larynx stage III (T3N0M0) squamous cell ca keratinized well differentiatedis presented. The treatment undergoes with total laryngectomy.Keywords: Laryngeal cancer, carcinoma, laryngectomy, tobacco

Computer Based Correlation of the Texture of P63 Expressed Nuclei with Histological Tumour Grade, in Laryngeal Carcinomas

Background. P63 immunostaining has been considered as potential prognostic factor in laryngeal cancer. Considering that P63 is mainly nuclear stain, a possible correlation between the texture of P63-stained nuclei and the tumor’s grade could be of value to diagnosis, since this may be related to biologic information imprinted as texture on P63 expressed nuclei.Objective. To investigate the association between P63 stained nuclei and histologic grade in laryngeal tumor lesions.Methods. Biopsy specimens from laryngeal tumour lesions of 55 patients diagnosed with laryngeal squamous cell carcinomas were immunohistochemically (IHC) stained for P63 expression. Four images were digitized from each patient’s IHC specimens. P63 positively expressed nuclei were identified, the percentage of P63 expressed nuclei was computed, and 118 textural, morphological, shape, and architectural features were calculated from each one of the 55 laryngeal lesions. Data were split into the low grade (21 grade I lesions) and high grade (34 grade II and grade III lesions) classes for statistical analysis.Results. With advancing grade, P63 expression decreased, P63 stained nuclei appeared of lower image intensity, more inhomogeneous, of higher local contrast, contained smaller randomly distributed dissimilar structures and had irregular shape.Conclusion. P63 expressed nuclei contain important information related to histologic grade.