Antitumor activity of 7RH, a discoidin domain receptor 1 inhibitor, alone or in combination with dasatinib exhibits antitumor effects in nasopharyngeal carcinoma cells.

Dysregulation of the discoidin domain receptors (DDRs) has been implicated in the development of numerous types of tumors, including head and neck cancer, and nasopharyngeal, breast, ovarian and esophageal carcinomas. Furthermore, agents that inhibit DDR1 activity are hypothesized to be useful for the treatment of nasopharyngeal carcinoma (NPC). The aim of the present study was to evaluate the effect of the DDR1 inhibitory (3-(2-(pyrazolo(1,5-a)pyrimidin-6-yl)-ethynyl)benzamide compound, 7RH, in NPC cells both in vitro and in vivo, and its effect when used in combination with dasatinib, a SRC family kinase (SFK) inhibitor. The effects of 7RH alone or in combination with dasatinib on cell viability were assessed using MTT assays and apoptosis was detected by flow cytometry. In addition, western blotting was performed to analyze the relative protein expression levels of cell cycle-associated genes in human NPC cell lines (CNE1, CNE2, HONE1 and SUNE1). Cell migration was also assessed using cell adhesion assays. Furthermore, tumor xenografts of CNE2 NPC cells were established in nude mice and the growth inhibitory effects of 7RH treatment alone or in combination with dasatinib were evaluated. Finally, knockdown of DDR1 protein expression was achieved by transfection of CNE2 cells with DDR1-specific small interfering RNA. Treatment with 7RH effectively suppressed the proliferation and induced the apoptosis of NPC cells. In addition, the Janus kinase 1 (JAK1)/signal transducer and activator of transcription (STAT3) signaling pathway was downregulated by 7RH, whereas the activities of the Ras/Raf/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK) and phosphoinositide 3-kinase (PI3K)/AKT signaling pathways were upregulated in response to 7RH treatment. Furthermore, the expression levels of phosphorylated SRC were increased in NPC cells treated with 7RH; thus indicating that SRC exhibits a vital function in the resistance of NPC cells to 7RH via activation of the PI3K/AKT signaling pathway. The results of the present study indicate that DDR1 and SFK inhibition may present a potential therapeutic strategy for patients with NPC.

The vocal aerodynamic change in female patients with muscular tension dysphonia after voice training.

OBJECTIVE: To investigate the changes of vocal aerodynamics indicators after voice training in female patients with muscular tension dysphonia (MTD). METHODS: Twenty-one female MTD patients (before voice training and 12 weeks after voice training) and 20 female volunteers with normal voices (the control group) received vocal aerodynamic analysis. Parameters included subglottal pressure (SGP), aerodynamic power (AP), mean expiratory airflow (MEA), and maximum phonation time (MPT) were recorded and analyzed by phonatory aerodynamic system. RESULTS: Before voice training, the median SGP and mean AP were higher than control group, whereas median MPT was shorter, and these differences were statistically significant. After 12 weeks of voice training, the median SGP and mean AP were decreased and the median MPT was increased compared with the measurements obtained before training, and these differences were statistically significant. The differences of median SGP, mean AP, mean MEA, and median MPT between MTD after 12 weeks of training and control group were not statistically significant. CONCLUSION: Voice training is an effective treatment for MTD patients. Aerodynamic analysis can effectively evaluate the vocal functional status of MTD patients before and after training, which is beneficial for the treatment efficacy evaluation.

[Mechanism of radiosensitization of 3-methyladenine in human hypopharynx cancer cells].

OBJECTIVE: To investigate the radiosensitizing effect and its mechanism of 3-MA in human hypopharynx cancer cells. METHODS: 5 mmol/L of 3-MA combined with 2 Gy or 4 Gy of X-ray was utilized to deal with Fadu cells, and the cell livability (cloning efficiency) and DNA lesion severity (tail moment) of each groups was examined by clonogenic survival assay and comet assay, then differences were compared between groups by independent-sample T test. Fadu cells were then treated with different dose of 3-MA (1, 2, 5, 10 mmol/L), the alteration of cell cycle distribution was detected by flow cytometer, and differences among groups were analyzed through one-way analysis of variance. The expression of p62 and cyclinB1 in each group was examined by western blot. RESULTS: The livability and DNA lesion severity of cells treated with 3-MA alone showed no notable variation. Compared with non-3-MA groups, the cloning efficiency of cells treated with 3-MA decreased much more after irradiated with 2 Gy or 4 Gy of X-ray (t = 13.41 or 13.98, P < 0.001), and the cells showed a more serious DNA lesion (t = 7.07 or 6.91, P < 0.001). The G2/M percentages of cells in the control group and groups treated with 1, 2, 5, 10 mmol/L of 3-MA were 17.10 ± 1.20, 23.30 ± 2.3, 39.90 ± 3.12, 58.47 ± 1.65, 76.13 ± 3.51 and differences among groups were statistically significant (F = 278.4, P < 0.05). The expression of p62 in cells treated with 3-MA showed a dose-dependent increase, while cyclinB1 showed a dose-dependent decrease. CONCLUSIONS: The autophagy inhibitor 3-MA could enhance radiosensitivity of human hypopharynx cancer cells by inducing G2/M arrest and enhancing irradiation-induced DNA damage.

[Application of narrow band imaging endoscopy in the diagnosis of nasopharyngeal lesions].

OBJECTIVE: To evaluate the value of narrow band imaging (NBI) endoscopy in the diagnosis of nasopharyngeal lesions. METHODS: Between December 2009 and April 2010, a total of 124 patients with nasopharyngeal lesions were examined with electronic nasopharyngolaryngoscope equipped with the white light mode and NBI mode. The biopsies of nasopharyngeal lesions were done in all patients. The characteristics of morphologies of mucosa and mucosal superficial vessels of each lesion under NBI mode were evaluated. RESULTS: Of all cases, there were 1 of papilloma, 87 of lymphoid follicular hyperplasia and chronic inflammation, 11 of nasopharyngeal cysts, and 25 of nasopharyngeal carcinoma. According to the pathological results, the morphologies of nasopharyngeal lesions under NBI mode were quite different. The color depth of the mucosa could be divided into four types: light red (+), dark red (++), prunosus (+++), and blue or blue black (++++). Under NBI, the color depths were (+) in papilloma, (++) in nasopharyngeal cysts, and (+++) in lymphoid follicular hyperplasia and chronic inflammation, without abnormal vessels. The color depths were (+++)-(++++) in nasopharyngeal cancer, with abnormal vessels. CONCLUSION: NBI has a potential ability to predict pathological results of nasopharyngeal lesions.

Inhibitor of differentiation 1 contributes to head and neck squamous cell carcinoma survival via the NF-kappaB/survivin and phosphoinositide 3-kinase/Akt signaling pathways.

PURPOSE: A key issue in cancer is apoptosis resistance. However, little is known about the transcription factors that contribute to cellular survival of head and neck squamous cell carcinoma (HNSCC). EXPERIMENTAL DESIGN: Three batches (54, 64, and 38) of HNSCC specimens were used for cellular and molecular analyses to determine the major molecular signaling pathways for cellular survival in HNSCC. Animal models (cell culture and xenografts) were used to verify the importance of apoptosis resistance in HNSCC. RESULTS: Inhibitor of differentiation (Id) family member, Id1, was significantly upregulated in clinical HNSCC specimens and acted to protect keratinocytes from apoptosis. Transfection of HNSCC cells with Id1 in vitro induced the phosphorylation of Akt (p-Akt) via phosphoinositide 3-kinase and increased the expression of survivin via NF-kappaB. Blockage of both pathways by specific inhibitors (LY294002 and IkappaBalphaM, respectively) abrogated Id1-induced cell survival of keratinocytes. In vivo studies showed that increased expression of Id1 allowed nontumorigenic keratinocytes (Rhek-1A) to become tumorigenic in nude mice by increased expression of survival genes such as p-Akt and survivin. More importantly, short interfering RNA for Id1 significantly reduced HNSCC tumor volume of HNSCC in xenograft studies. Analysis of clinical data verified the importance of the Id1 downstream molecule, survivin, in the prognosis of HNSCC patients. CONCLUSIONS: The above data, taken together, suggest that Id1 and its downstream effectors are potential targets for treatment of HNSCC because of their contribution to apoptosis resistance.

[Repair of pharynx with acellular dermal matrix].

OBJECTIVE: To evaluate the clinical use of acellular dermal matrix (ADM) in the repair of pharynx. METHOD: From June 2003 to December 2004, 18 cases of laryngopharyngeal carcinoma or oropharyngeal carcinoma accepted treatment of surgery. Four of them had the tumor in the place of posterior wall of laryngopharynx or oropharynx. In the operation, the tumor was resected and the function of larynx was reserved, reconstruction of the posterior wall of laryngopharynx or oropharynx was made with ADM. Three of them had normal function of vocal cords and the tumor was in the place of posterior wall of pharynx and lateral wall of piriform fossa. These patients accepted resection of the tumor and the larynx was reserved. ADM was sutured to the prevertebral fascia to repair the pharynx, reinforcement was made with sternocleidomastoid muscle then. The other 11 patients had the tumor in the pharynx and cervico-esophagus and the vocal cords were fixed. These cases accepted total resection of the larynx and laryngopharynx. ADM was sutured to the prevertebral fascia to repair the posterior wall of laryngopharynx, musculo-cutaneous flap of pectoralis major muscle was made to repair the anterior and lateral walls. All these 18 patients accepted radiotherapy after the operation, the dosage was 60-70 Gy. RESULTS: Among all these 18 cases, no rejection of ADM or formation of pharynx fistula happened after the operation. The allograft became normal mucosa finally. Two patients had subcutaneous infection. After the treatment of dressing change and antibiotics, the wound healing was satisfying. All the 18 patients had resumption of oral diet finally but 3 of them had light mis-swallowing. Seven patients whose larynx were reserved had the trachea cannula pulled out The follow-up periods after the operation lasted 12 months to 30 months. The middle follow-up period was 19. 38 months. Neither rejection nor scar formation of the allograft happened. The follow-up of 11 patients lasted more than 18 months, 3 of them had relapse of tumor. After the second operation 1 patient was still alive and 2 patients died of distant metastasis. CONCLUSION: ADM can be easily obtained. Its histocompatibility were perfect The operative procedure were easy to perform. The complications of donor part can be totally avoided. The combined use of ADM and musculo-cutaneous flap of pectoralis major muscle or other neck tissue was safe and effective in the reconstruction of pharynx.

[Analysis of surgical treatment of nasopharyngeal angiofibroma].

OBJECTIVE: To study the individual surgical treatment of nasopharyngeal angiofibroma. METHODS: Between Feb. 1989 and Oct. 2004, different kinds of surgical approaches in 51 patients with angiofibromas were retrospectively analysed. Using Fisch stage: thirty three cases were in stage I and II, 22 cases via palatal approach, 11 cases via endoscopic surgery;18 cases were in stage IIN and IV, 9 cases via lateral rhinotomy approach, 4 cases via midfacial degloving approach + Lefort I approach or modified maxillotomy, 5 cases via craniofacial approach. Twenty three cases accepted the embolization of the artery which feed the tumor, 7 cases in stage IV, 8 cases in stage IlI, 5 cases in stage II, 3 cases in stage I. RESULTS: The average of blood loss was 1010 ml in operation but it became 200 - 870 ml (the average of blood loss was 485 ml) when had a selective preoperative embolization. The blood loss of none selective preoperative embolization was 500 - 3500 ml (the average of blood loss was 1600 ml), t = 7. 48, P < 0.05, the average of blood loss of selective preoperative embolization is less than the average of blood loss of none selective preoperative embolization. After 9 to 48 months of operation (the average time was 26.4 months), 8 cases recurred one time and 2 cases recurred two times. CONCLUSIONS: The preoperative embolization could reduce the amount of bleeding during operation. In patients who are in stage I and II, searching endoscopic surgery could avoid the effect on craniofacial development which traditional surgical procedure did. For patient who are in stage III and IV, midfacial degloving approach and Lefort I approach or modified maxillotomy are not only good for exclusive resection of the tumor but also have less effect on articulation and beautiful cosmetic outlook.

[Endoscopic thyroid gland surgery without carbon dioxide neck insufflation].

OBJECTIVE: To evaluate endoscopic thyroid gland surgery without carbon dioxide neck insufflation. METHODS: Sixteen patients with thyroid gland benign tumor, aged from 23-62 years, were selected and treated with endoscopic surgery. Eight cases received whole thyroid lobectomy and the other eight cases received partial thyroid lobectomy, the procedure was through a wall method or a single 2.0-3.0 cm horizontal skin incision. Among the 16 cases, 6 were thyroid adenoma and 10 were nodular goiter. RESULTS: All patients received endoscopic surgery, 2 cases with sub-clavicle approach and 14 cases with a single 2.0-3.0 cm horizontal skin incision approach. The recurrent laryngeal nerve and parathyroid glands were easily identified and preserved during operation, the operating time lasted from 1.1 hours to 4.0 hours, no complication occurred, the postoperative cosmetic outcome was excellent. CONCLUSIONS: The results indicated that gasless technique is feasible and safe, so it suggested that the video-assisted thyroid surgery without carbon dioxide neck insufflation could be an alternative for thyroid gland surgery.

[Relationship between expression of survivin and prognosis in human laryngeal squamous cell carcinoma].

BACKGROUND & OBJECTIVE: A novel inhibitor of apoptosis Survivin has recently been found in many common human cancers but not in normal tissues. Its potential distribution in human laryngeal squamous cell carcinoma (HLSCC) and its implication for inhibition of apoptosis were not yet very clear. This study was designed to investigate the expression of survivin and the probabilities of using it as a new indicator of the prognosis of HLSCC. METHODS: HLSCC samples from 71 patients treated in Department of Otorhinolaryngology, Second Affiliated Hospital of Sun Yat-sen University between 1995 and 1998 were analyzed for survivin expression by immunohistochemistry method. The patients were followed-up for long term and the relationship between survivin expression and clinical data including age, gender, classification, clinical type, pathological type, lymph node metastasis, distant metastasis, and prognosis were analyzed using software SPSS11.0. RESULTS: The expression rate was 50.7% (36/71) in all 71 cases. The expression rate was 81.8% (9/11) in the cases with lymph nodes metastasis, which was higher than that in the cases without lymph nodes metastasis (45%, 27/60), with significant difference (P=0.025). Univariate analysis revealed that classification,type and survivin expression were significantly related to the prognosis of HLSCC. P value were 0.0001, 0.009, and 0.0008, respectively. Multivariate analysis demonstrated that the patients with positive survivin had significant shorter survival (63.5 months) than those with negative survivin (84.0 months) (P=0.006) except the effect of classification and type on the prognosis of HLSCC. CONCLUSION: Survivin expressed in HLSCC especially in the cases with lymph nodes metastasis. Its expression may related to the prognosis of HLSCC.