Downregulation of RBM17 enhances cisplatin sensitivity and inhibits cell invasion in human hypopharyngeal cancer cells.

Most of advanced hypopharyngeal squamous cell carcinoma (HSCC) are resistant to chemotherapy, and there is still lack of effective treatment for HSCC now. The present study aimed to investigate whether downregulation of RNA-binding motif protein 17 (RBM17) could enhance cisplatin sensitivity and inhibit cell invasion in HSCC and the underlying mechanism. We observed that RBM17 was upregulated in tumor tissues and associated with poor progression. Treatment of FaDu cells with cisplatin increased RBM17 expression in mRNA levels. Downregulation of RBM17 enhanced cisplatin-mediated inhibition of FaDu cells. In addition, downregulation of RBM17 effectively suppressed tumor cell migration and invasion through the reversion of epithelial-mesenchymal transition. Moreover, downregulation of RBM17 could significantly slow tumor growth in FaDu xenograft tumor model. Liquid chromatography-mass spectrometry/mass spectrometry detection and independent PRM analysis showed that 21 differentially expressed proteins were associated with the downregulation of RBM17. Taken together, our study implied that downregulation of RBM17 could serve as a novel approach to enhance cisplatin sensitivity in HSCC.

Clinicopathological significance of SOX4 and epithelial-mesenchymal transition markers in patients with laryngeal squamous cell carcinoma.

OBJECTIVE: Sex-determining region-Y-related high-mobility-group box 4 (SOX4) is associated with the metastasis and prognosis of many cancer types. However, studies on the role of SOX4 in laryngeal squamous cell carcinoma (LSCC) are few, and hence the mechanism is unclear. Epithelial-mesenchymal transition (EMT) allows neoplastic cells to gain the plasticity and motility required for tumor progression and metastasis. This study aimed to analyze the relationship between SOX4 and EMT, and their relationship with clinicopathological factors and related prognosis. METHODS: Immunohistochemical staining was used to detect the positive expression of SOX4 protein, EMT-related transcription factor protein, and related marker protein in 127 LSCC tissue samples. At the same time, data on various parameters of clinical pathology and postoperative survival were collected. RESULTS: The positive expression rate of SOX4 and Slug in LSCC was related to pathological differentiation, Lymph node metastasis (LNM), and pathological TNM of a tumor. The expression rates of ZEB1, E-cadherin, N-cadherin, and ß-catenin in LSCC correlated with LNM and pTNM. The expression of SOX4, combined expression of SOX4 and ZEB1, and LNM were independent prognostic factors for the total survival time of patients with LSCC. CONCLUSIONS: In summary, SOX4 was vital in the LSCC EMT process, which might be mediated by transcription factor ZEB1. SOX4 and ZEB1 might serve as potential biomarkers of metastasis and prognosis, as well as promising therapeutic targets of LSCC.

Clinicopathological significance of SOX4 and epithelial-mesenchymal transition markers in patients with laryngeal squamous cell carcinoma.

BACKGROUND AND AIM: Sex-determining region-Y-related high-mobility-group box 4 (SOX4) is associated with the metastasis and prognosis of many cancer types. However, studies on the role of SOX4 in laryngeal squamous cell carcinoma (LSCC) are few, and hence the mechanism is unclear. Epithelial-mesenchymal transition (EMT) allows neoplastic cells to gain the plasticity and motility required for tumor progression and metastasis. This study aimed to analyze the relationship between SOX4 and EMT, and their relationship with clinicopathological factors and related prognosis. METHODS: Immunohistochemical staining was used to detect the positive expression of SOX4 protein, EMT-related transcription factor protein, and related marker protein in 127 LSCC tissue samples. At the same time, data on various parameters of clinical pathology and postoperative survival were collected. RESULTS: The positive expression rate of SOX4 and Slug in LSCC was related to pathological differentiation, lymphatic invasion, and pathological tumor node metastasis (TNM) of a tumor. The expression rates of ZEB1, Twist, E-cadherin, N-cadherin, and ß-catenin in LSCC correlated with lymphatic invasion and pathological tumor node metastasis. The expression of SOX4, combined expression of SOX4 and ZEB1, and lymphatic invasion were independent prognostic factors for the total survival time of patients with LSCC. CONCLUSIONS: In summary, SOX4 was vital in the LSCC EMT process, which might be mediated by transcription factor ZEB1. SOX4 and ZEB1 might serve as potential biomarkers of metastasis and prognosis, as well as promising therapeutic targets of LSCC.

Clinical significance of USP7 and EZH2 in predicting prognosis of laryngeal squamous cell carcinoma and their possible functional mechanism.

BACKGROUND: Laryngeal squamous cell carcinoma (LSCC) is one of the most prevalent cancer types in the world. The ubiquitin specific protease 7 (USP7), a kind of deubiquitylating enzyme, has been reported to play multifaceted roles in different tumor types. EZH2 has been found to be highly expressed in various malignantcells and high expression of EZH2 is closely related to tumor growth infiltration, lymph node involvement, clinical stage, and poor prognosis. The aim of this study was to investigate the expression and function of USP7 and EZH2 in LSCC. CASE PRESENTATION: Immunohistochemical staining and histochemical staining were performed to explore the expression of USP7 and EZH2 in both LSCC tissues and adjacent normal laryngeal tissues. Chi-square test, univariate analysis, and multivariate analysis were conducted to statistically evaluate the clinical significance of USP7 and EZH2. CONCLUSIONS: USP7and EZH2 affects LSCC evolution; USP7 and EZH2 were upregulated in LSCC tissues, which can serve as independent prognostic predictors, and potential therapeutic targets for LSCC.

[Long chain non-coding RNA MALAT-1 gene knockdown inhibits growth and migration and promotes apoptosis of human laryngeal squamous cell carcinoma Hep-2 cells in vitro].

OBJECTIVE: To investigate the effect of knocking down long chain non-coding RNA MALAT-1 gene on the biologicalbehaviors of human laryngeal squamous cell carcinoma Hep-2 cells. METHODS: With immortalized nasopharyngeal epithelial(NPE) cell line NP-69 as the reference, MALAT1 expression in FaDu, Hep-2 and nasopharyngeal carcinoma CNE-2Z cells weredetected using real-time PCR. Hep-2 cells were transfected with shmalat1 lentivirus and the expression of MALAT1 wasdetected. MTT assay, flow cytometry, Transwell assay and M Atrigel invasiveness test were used to evaluate the effect ofMALAT-1 knockdown on the proliferation, cell cycle, cell apoptosis, migration, and invasiveness of Hep-2 cells. RESULTS: Compared with NP-69 cells, Hep-2 cells, FaDu cells, and CNE-2Z cells all showed significantly increased MALAT-1expression. In Hep-2 cells, knockdown of MALAT-1 significantly inhibited the cell proliferation, increased the cell percentagein S phase (P < 0.01), decreased the cell percentage in G2/M phase (P < 0.01), and attenuated the migration and invasiveness of thecells. CONCLUSIONS: MALAT-1 is over-expressed in laryngeal squamous cell carcinoma, and knocking down MALAT-1 gene cansignificantly suppress the proliferation, invasion and migration and promotes apoptosis of the cancer cells.

Downregulation of RNA binding motif protein 17 expression inhibits proliferation of hypopharyngeal carcinoma FaDu cells.

RNA binding motif protein 17 (RBM17) is a protein-coding gene. The protein encoded by RBM17 is involved in the regulation of alternative splicing and is overexpressed in cancer. The present study aimed to determine the effect of RBM17-knockdown in hypopharyngeal carcinoma FaDu cells using the lentivirus-mediated shRNA method. Cell proliferation was detected by an MTT assay. Flow cytometry analysis was used to determine cell cycle distribution and apoptosis. The results of the present study demonstrated that RBM17 expression was significantly decreased in FaDu cells infected with lentivirus-shRNA. Knockdown of RBM17 expression by shRNA significantly reduced cell proliferation, augmented cell apoptosis and arrested cells at the G2/M phase in FaDu cells. The results of the present study indicate that RBM17 serves a notable role in cell proliferation, cell cycle progression and apoptosis of hypopharyngeal carcinoma cells.

[One-stage repair of pharyngeal defect using tongue flaps after resection of advanced stage hypopharyngeal neoplasm and laryngeal neoplasm].

OBJECTIVE: To study the effectiveness of one-stage repairing pharyngeal defect with the tongue flaps after resection of advanced stage hypopharyngeal neoplasm and laryngeal neoplasm. METHODS: Between June 2006 and March 2011, 20 patients with hypopharyngeal neoplasm (8 cases) and laryngeal neoplasm (12 cases) with advanced stage were treated. There were 19 males and 1 female, aged 47-78 years (mean, 62.8 years). All neoplasms were squamous cell carcinomas. The disease duration was 1-8.5 months (mean, 3.9 months). According to the standards of International Union Against Cancer (UICC, 1987), 12 cases were in stage III and 8 cases were in stage IV. The size of pharyngeal defect was 5 cm x 2 cm to 4 cm x 4 cm after resection of tumor. Defects were repaired by the whole base of the tongue flaps in 16 cases and by the horizontal base of the tongue flaps in 4 cases. The size of the flaps ranged from 5 cm x 2 cm to 4 cm x 4 cm. Postoperative radiotherapy and chemotherapy were regularly performed. RESULTS: The 20 tongue flaps were alive. Healing of incision by first intention was achieved in 18 cases and delayed healing in 2 cases because of subcutaneous fluid. The patients were followed up 12-63 months (mean, 36.7 months). The patients had normal feeding ability and tongue function. Of 20 cases, 12 died and 1 of local recurrence was alive with tumor. The 3-year survival rate was 69.2% (9/13). CONCLUSION: One-stage repair of pharyngeal defect with the tongue flaps after resection of hypopharyngeal neoplasm and laryngeal neoplasm can obtain good effectiveness because the tongue flap is easy-to-obtain and easy-to-survive, and has abundant blood supply.

[Surgery for T4 hypopharyngeal cancer and reconstruction after hypopharyngo-oesphagectomy].

OBJECTIVE: To explore the surgical treatment of hypopharyngeal and cervical esophageal cancers and the ways of reconstruction after hypopharyngo-oesphagectomy, and to evaluate their efficacy. METHODS: Twenty five patients with cancer of the laryngopharynx and cervical esophagus treated in our department between 1995 and 2007 were included in this study. Their clinical data were restrspectively analyzed. Among them, 17 cases had the tumor originated from the pyriform sinus, 3 of the posterior pharyngeal wall and 5 of the postcricoid region. Acording to the 2002 UICC criteria, all the tumors were stage T4, including 9 patients with cN0, 11 with cN1, and 5 with cN2 disease. The pharyngoesophageal defect reconstruction methods were as following: pharyngogastric anastomosis in 7 patients, free jejunal transplantion in 4, laryngotracheal flap in 8, and pectoralis major musculocutaneous flap in 6 patients. All patients were treated with modified and/or selective neck dissection. Among them, 8 cases received pre-operation radiotherapy, 17 received post-operative auxiliary radiotherapy. RESULTS: There was no operation death case in this group. All patients were followed up for 3 to 5 years. Three patients died in the first year. According to Kaplan-Meier analysis, the 1-year survival rate was 88.0%, 3-year survival rate was 48.0%, and 5-year survival rate was 28.0%. CONCLUSIONS: The use of primary repair of the defects of laryngopharynx and cervical esophagus expands the operative indication for cancers of the laryngopharynx and cervical esophagus, improves the survival rate and life quality of the patients. Regarding the repair method of choice, site of the tumor and size of the defect are the most important factors regarding choice of reconstruction method, while the patients' age and general condiction should also be considered to minimize the complications as more as possible.

Decoding task-based attentional modulation during face categorization.

Attention is a neurocognitive mechanism that selects task-relevant sensory or mnemonic information to achieve current behavioral goals. Attentional modulation of cortical activity has been observed when attention is directed to specific locations, features, or objects. However, little is known about how high-level categorization task set modulates perceptual representations. In the current study, observers categorized faces by gender (male vs. female) or race (Asian vs. White). Each face was perceptually ambiguous in both dimensions, such that categorization of one dimension demanded selective attention to task-relevant information within the face. We used multivoxel pattern classification to show that task-specific modulations evoke reliably distinct spatial patterns of activity within three face-selective cortical regions (right fusiform face area and bilateral occipital face areas). This result suggests that patterns of activity in these regions reflect not only stimulus-specific (i.e., faces vs. houses) responses but also task-specific (i.e., race vs. gender) attentional modulation. Furthermore, exploratory whole-brain multivoxel pattern classification (using a searchlight procedure) revealed a network of dorsal fronto-parietal regions (left middle frontal gyrus and left inferior and superior parietal lobule) that also exhibit distinct patterns for the two task sets, suggesting that these regions may represent abstract goals during high-level categorization tasks.

[Application of pectoralis major myocutaneous flap in repair of defect caused by resection of tonsillar cancer].

OBJECTIVE: To discuss and value the effects and the advantages of pectoralis major myocutaneous flap in repair of defect caused by resection of tonsillar cancer. METHODS: The data were retrospectively summarized from 10 patients with recurrent tonsillar cancer after radical radiotherapy from January 1998 to December 2005, including 7 cases of squamous cell carcinoma, 2 cases of undifferentiated carcinoma, and 1 case of adenoid cystic carcinoma. There were 8 males and 2 females, aged 43-68 years with an average of 58 years. All cases were classified as stages III and IV before radiotherapy according to staging standard of oropharyngeal cancer (International Union Against Cancer, 1997). The time of relapse was 6-32 months after radiotherapy. Recurrent tonsil cancer invaded tongue base, soft palate, posterior wall of pharyngeal, parapharyngeal space, and palate. Tumor size was from 4 cm x 2 cm to 8 cm x 5 cm. Seven cases were accompanied by lymph node metastasis. After carcinoma were completely resected and defects were reconstructed by pectoralis major myocutaneous flap of 7 cm x 5 cm-12 cm x 9 cm. The donor sites were sutured directly. RESULTS: After operation, pectoralis major myocutaneous flap completely survived in 9 cases. Partial necrosis of pectoralis major myocutaneous flap was found in 1 case; after treatment, the necrotic flap remained small pharyngeal defect. Incision at donor site healed by first intention in 10 cases. All patients showed satisfactory functions of respiratory, voice, and swallowing with no complication. Ten patients were followed up 2 years to 5 years and 8 months. The 3-year survival rate was 66.7% (6/9), and the 5-year survival rate was 20.0% (1/5). CONCLUSION: Pectoralis major muscle flap has a high survival rate, which is safe, reliable, easy-to-operate, and can repair larger defect. Pectoralis major myocutaneous flap is an ideal material in repair of defect caused by resection of recurrent tonsillar carcinoma after radiotherapy.

In situ measurement of solute transport in the bone lacunar-canalicular system.

Solute transport through the bone lacunar-canalicular system is believed to be essential for osteocyte survival and function but has proved difficult to measure. We report an approach that permits direct measurement of real-time solute movement in intact bones. By using fluorescence recovery after photobleaching, the movement of a vitally injected fluorescent dye (sodium fluorescein) among individual osteocytic lacunae was visualized in situ beneath the periosteal surface of mouse cortical bone at depths up to 50 microm with laser scanning confocal microscopy. Transport was analyzed by using a two-compartment mathematical model of solute diffusion that accounted for the characteristic anatomical features of the lacunar-canalicular system. The diffusion coefficient of fluorescein (376 Da) was determined to be 3.3 +/- 0.6 x 10(-6) cm2/sec, which is 62% of its diffusion coefficient in water and is similar to diffusion coefficients measured for comparably sized molecules in cartilage. The diffusion of fluorescein in bone is also consistent with the presence of an osteocyte pericellular matrix whose structure resembles that proposed for the endothelial glycocalyx [Squire, J. M., Chew, M., Nneji, G., Neal, C., Barry, J. & Michel, C. (2001) J. Struct. Biol. 136, 239-255]. To our knowledge, this is the first instance where the dynamics of molecular movement has been measured directly in the bone lacunar-canalicular system. This in situ imaging approach should also facilitate the analysis of convection-based transport mechanisms in bones of living animals.

Mechanotransduction and strain amplification in osteocyte cell processes.

A paradox in bone tissue is that tissue-level strains due to animal and human locomotion are too small to initiate intracellular chemical responses directly. A model recently was proposed to resolve this paradox, which predicts that the fluid flow through the pericellular matrix in the lacunar-canalicular porosity due to mechanical loading can induce strains in the actin filament bundles of the cytoskeleton that are more than an order of magnitude larger than tissue level strains. In this study, we greatly refine this model by using the latest ultrastructural data for the cell process cytoskeleton, the tethering elements that attach the process to the canalicular wall and their finite flexural rigidity EI. We construct a much more realistic 3D model for the osteocyte process and then use large-deformation "elastica" theory for finite EI to predict the deformed shape of the tethering elements and the hoop strain on the central actin bundle. Our model predicts a cell process that is 3 times stiffer than in a previous study but hoop strain of >0.5% for tissue-level strains of >1,000 microstrain at 1 Hz and >250 microstrain at frequencies >10 Hz. We propose that this strain-amplification model provides a more likely hypothesis for the excitation of osteocytes than the previously proposed fluid-shear hypothesis.

[Different surgical management of tumors in the anterior skull base: a report of 10 cases].

OBJECTIVE: To study the different surgical approach for the resection of tumors in the anterior skull base and to introduce an approach of the frontal mid-line incision with the nasal pyramid over-turned downward. METHOD: A review of 10 tumors in the anterior skull base employed unlike operative approach was presented. Of the 10 cases, 6 cases were benign tumors and 4 cases were malignant tumors. Two cases underwent the combined craniofacial approach, 3 cases the subfrontal approach, 3 cases the lateral rhinotomy approach, 2 cases the approach of the frontal mid-line incision with the nasal pyramid over-turned downward. RESULT: All tumors were totally resected. All patients were followed up from 1 to 5 years. In the benign tumors group, 6 cases achieved good results and no recurrence. Of the 4 cases with malignant tumors, 1 cases were followed up 1 year and is alive, 2 cases survived after 3 years and 1 case died after 5 years. Three cases died from local recurrence or invaded intra-cranial. CONCLUSION: According to the site, range and pathology of the skull base tumors, the best operative approach should been selected. The approach of the frontal mid-line incision with the nasal pyramid over-turned downward is closer to tumor and easy to repair duramater, has fine exposure of the anterior skull base, minimal retraction of the frontal lobe, and has better clinically worthy.

[Pearson technique in the treatment of advanced laryngeal cancer].

OBJECTIVE: To seek for a better method of treating the advanced laryngeal cancer. METHOD: Twenty-eight cases of advanced laryngeal cancer were treated with near-total laryngectomy(Pearson technique), including 11 glottic cancers(III stage, 8 cases: IV stage, 3 cases), 17 supraglottic cancers (III stage, 13 cases; IV stage, 4 cases). In the same time 26 patients had neck node dissections. All the cases had speech function reconstructions. RESULT: The speech function was obtained in 27/28(96.4%). All cases had good swallowing, 3-year survival rate is 78.6%, 5-year survival rate is 68.4%. CONCLUSION: Near-total laryngectomy can be used in most cases that might be treated with traditional total laryngectomy in the past. It can provide satisfied survival rate, good swallowing, better speech function, but breath function.

Mechanotransduction and flow across the endothelial glycocalyx.

In this inaugural paper, we shall provide an overview of the endothelial surface layer or glycocalyx in several roles: as a transport barrier, as a porous hydrodynamic interface in the motion of red and white cells in microvessels, and as a mechanotransducer of fluid shearing stresses to the actin cortical cytoskeleton of the endothelial cell. These functions will be examined from a new perspective, the quasiperiodic ultrastructural model proposed in Squire et al. [Squire, J. M., Chew, M., Nneji, G., Neal, C., Barry, J. & Michel, C. (2001) J. Struct. Biol. 136, 239-255] for the 3D organization of the endothelial surface layer and its linkage to the submembranous scaffold. We shall show that the core proteins in the bush-like structures comprising the matrix have a flexural rigidity, EI, that is sufficiently stiff to serve as a molecular filter for plasma proteins and as an exquisitely designed transducer of fluid shearing stresses. However, EI is inadequate to prevent the buckling of these protein structures during the intermittent motion of red cells or the penetration of white cell microvilli. In these cellular interactions, the viscous draining resistance of the matrix is essential for preventing adhesive molecular interactions between proteins in the endothelial membrane and circulating cellular components.

[A clinical evaluation on supracricoid partial laryngectomy with laryngoplasty].

OBJECTIVE: To evaluate the results of supracricoid partial laryngectomy with laryngoplasty in treating laryngeal cancer and reconstructing its functions. METHODS: One hundred and fifty-nine patients receiving subtotal laryngectomy from 1993 to 1999 were analysed. 4 kinds of operations were performed for them. 46 cases underwent supracricoid partial laryngectomy with the reconstruction of laryngeal function by pedicled flaps. RESULTS: Among the 46 cases receiving supracricoid partiallaryngectomy with laryngoplasty, aspiration did not occur in 40 cases. The decannulation rate was 91.3%, 3, 5 year survival rates were 84.8%, 75%. 41 cases resumed enjoyed satisfactory phonation. CONCLUSION: This form of laryngeal reconstruction in supraericoid partial laryngectomy with laryngoplasty is helpful to restore laryngeal function. It can prevent aspiration and improve the decannulation rate as well as the quality of life in partial laryngectomy patients.