RESUMO
OBJECTIVES: To assess and compare the effectiveness of various treatment approaches for laryngeal contact granulomas (LCG). METHODS: A retrospective analysis was conducted on a cohort of 45 patients diagnosed with LCG at the Second Affiliated Hospital of Xi'an Jiaotong University from October 2017 to May 2023. Based on the treatment modalities administered, patients were categorized into three groups: acid suppression alone, hormone injection combined with acid suppression, and surgery combined with acid suppression. Subsequently, the study compared differences in treatment efficacy and average healing time among these three groups, using various indicators. RESULTS: The findings indicate that the granuloma size in LCG patients with hoarseness (0.126, 95% CI 0.087-0.288) was significantly greater compared to LCG patients without hoarseness (0.047, 95% CI 0.014-0.083) (P = 0.001). However, there were no significant variations in age, morphology (unlobulated/lobulated), laterality ratio (left/right), sex ratio (male/female), history of tracheal intubation (non-intubation/intubation), and RFS score (RFS > 7/RFS ≤ 7) (P > 0.05), regardless of the presence of hoarseness symptoms. At the treatment observation endpoint of 3 months, the curative ratio in the group receiving hormone injection combined with acid suppression was found to be significantly higher compared to the group receiving acid suppression alone (P = 0.018). In addition, the average healing time of patients in the hormone injection combined with acid suppression group was notably shorter than that of the acid suppression alone group (P = 0.007). CONCLUSIONS: The combination of hormonal injections and acid suppression may enhance the curative ratio and expedite the healing time of LCG.
Assuntos
Granuloma Laríngeo , Rouquidão , Humanos , Masculino , Feminino , Estudos Retrospectivos , Rouquidão/etiologia , Rouquidão/terapia , Granuloma Laríngeo/cirurgia , Granuloma , HormôniosRESUMO
OBJECTIVE: To analyze the risk factors for recurrence of laryngeal amyloidosis (LA). METHODS: The clinical data of patients with LA admitted in the Otolaryngology Head and Neck Surgery Department of the Second Affiliated Hospital of Xi'an Jiaotong University from August 2009 to June 2022 were analyzed retrospectively; then, the risk factors for recurrence and their impacts on the recurrence time were also analyzed. RESULTS: Of the 44 patients with LA, the majority (38 cases, 86.4%) only involved one anatomical region and the others (6 cases, 13.6%) involved two laryngeal regions concurrently. Overall, the glottic region was the most commonly affected area (28 cases, 63.6%), followed by the supraglottic region (16 cases, 36.4%) and subglottic region (6 cases, 13.6%). In addition, all the lesions were categorized as isolated nodule (31.8%), submucosal localized deposition (52.3%), and submucosal diffuse deposition (15.9%) according to their morphologies under electronic laryngoscope. Finally, six patients (13.6%) had recurrence after operation with a median recurrence time of 24.5 months, and subglottic involvement was confirmed to be an independent risk factor for recurrence of LA by univariate and multivariate logistic regression analyses (P < 0.05). Meanwhile, the patients with subglottic involvement presented as submucosal diffuse deposition had a considerable shorter recurrence time (t = 5.759, P = 0.005). CONCLUSION: The subglottic involvement is an independent risk factor for recurrence of LA.
Assuntos
Amiloidose , Neoplasias Laríngeas , Laringe , Humanos , Neoplasias Laríngeas/cirurgia , Estudos Retrospectivos , Laringe/patologia , Fatores de RiscoRESUMO
OBJECTIVE: This study aimed to analyze the characteristics of laryngopharyngeal reflux (LPR) by using narrow band imaging (NBI) endoscopy. STUDY DESIGN: A prospective study. SETTING: A large-volume practice with tertiary care providers. METHODS: A total of 67 patients with suspected LPR who underwent 24-hour multichannel intraluminal impedance-pH monitoring were included from June 2020 to March 2022. Manifestations of NBI endoscopy included submucosal clustered brownish microvessels (CBMs), spotted brownish microvessels, and no special microvessels; the latter 2 formed the non-CBM group. The manifestations of all patients and their changes were observed after 8 weeks of proton pump inhibitor and symptomatic treatment for patients with LPR, and symptomatic treatment for patients without LPR. RESULTS: According to the results of 24-hour multichannel intraluminal impedance-pH monitoring, the incidence of submucosal CBMs was significantly higher in patients with LPR (30 cases) than in those without LPR (37 cases, P < .001), particularly in the posterior cricoid area (P < .001). Besides Reflux Finding Score, the incidence of signs such as subglottic edema and vocal fold edema was significantly higher in the CBM group than the non-CBM group (P < .05). Finally, 22 patients with LPR (91.7%) and only 2 patients without LPR (28.6%) underwent a transformation from CBMs to spotted brownish microvessels after continuous medication for 8 weeks in the CBM group (χ2 = 15.916, P < .001), while no significant change was observed in patients with or without LPR in the non-CBM group (P > .05). CONCLUSION: Submucosal CBMs in the posterior cricoid area under NBI endoscopy may be a characteristic of LPR.
Assuntos
Refluxo Laringofaríngeo , Humanos , Refluxo Laringofaríngeo/diagnóstico , Imagem de Banda Estreita , Estudos Prospectivos , Monitoramento do pH Esofágico , Endoscopia , EdemaRESUMO
OBJECTIVE: This study aimed to analyze the status of hypoxia in non-rapid eye movement (NREM) sleep in children with otitis media with effusion (OME). METHODS: A total of 232 children with OME and/or adenotonsillar hypertrophy were enrolled in this retrospective study between August 2020 and November 2021. Polysomnographic monitoring was carried out, and the differences in polysomnographic results between the experimental group (children with OME and adenotonsillar hypertrophy) and control group (children with adenotonsillar hypertrophy only) were compared. RESULTS: The lowest oxygen saturation level during sleep was significantly lower in the experimental group (n = 36) than in the control group (n = 196). However, the apnea-hypopnea index, respiratory disorder index, apnea index, obstructive apnea index, obstructive apnea-hypopnea index, and mixed apnea-hypopnea index were significantly higher in the experimental group than in the control group. More importantly, the apnea-hypopnea index, the oxygen desaturation index, oxygen desaturation events, the average heart rate during NREM sleep, and the NREM stage in total sleep time were also significantly higher in the experimental group than in the control group. CONCLUSIONS: Hypoxia during NREM sleep may affect the severity of OME in children.
Assuntos
Otite Média com Derrame , Apneia Obstrutiva do Sono , Criança , Humanos , Otite Média com Derrame/complicações , Apneia Obstrutiva do Sono/complicações , Estudos Retrospectivos , Movimentos Oculares , Sono/fisiologia , Hipóxia , Hipertrofia , OxigênioRESUMO
OBJECTIVE: To investigate the effect of Helicobacter pylori (HP) eradication therapy on salivary pepsin concentration in laryngopharyngeal reflux (LPR) patients with HP infection. MATERIALS AND METHODS: A total of 477 patients with suspected LPR were enrolled from June 2020 to September 2021. Reflux symptom index, reflux finding score, the positive rates and disintegrations per minute values of HP infection detected by 14C urea breath test and salivary pepsin concentrations analyzed using enzyme-linked immunosorbent assay were compared in LPR patients and non-LPR patients with or without HP infection. HP-positive patients were treated with HP eradication therapy while HP-negative patients with PPI therapy. RESULTS: The scores of nagging cough (0.88 vs. 0.50, P = 0.035), erythema or hyperemia (1.93 vs. 1.78, P = 0.035) and vocal fold edema (1.04 vs. 0.85, P = 0.025) were higher in the LPR (+) Hp (+) subgroup than in LPR (+) Hp (-) subgroup. The concentrations of salivary pepsin in the Hp (+) subgroup were higher than in the Hp (-) subgroup either in LPR patients (75.24 ng/ml vs. 61.39 ng/ml, P = 0.005) or the non-LPR patients (78.42 ng/ml vs. 48.96 ng/ml, P = 0.024). Compared to baseline (before treatment), scores of nagging cough (0.35 vs. 0.84, P = 0.019) and erythema or hyperemia (1.50 vs. 1.83, P = 0.039) and the concentrations of salivary pepsin (44.35 ng/ml vs. 74.15 ng/ml, P = 0.017) in LPR patients with HP infection decreased after HP treatment; yet, this was not observed for the LPR patients without HP infection treated with PPI only (P > 0.05). CONCLUSION: HP infection may aggravate the symptoms and signs of LPR patients, partly by increasing their salivary pepsin concentration.
Assuntos
Helicobacter pylori , Hiperemia , Refluxo Laringofaríngeo , Tosse , Humanos , Refluxo Laringofaríngeo/complicações , Refluxo Laringofaríngeo/diagnóstico , Refluxo Laringofaríngeo/tratamento farmacológico , Pepsina A , Saliva , UreiaRESUMO
Emerging studies have demonstrated that long noncoding RNAs (lncRNAs) play essential roles in tumorigenesis. However, the role and function of lncRNAs in hypopharyngeal squamous cell carcinoma (HSCC) have not been completely elucidated. The present study explored the function of a novel lncRNA, RP11156L14.1, in HSCC. RP11156L14.1 was revealed to be highly expressed in HSCC tissues and cell lines. Knockdown of RP11156L14.1 inhibited proliferation, migration, and invasion in HSCC cells. Furthermore, RP11156L14.1 regulated epithelialmesenchymal transition (EMT) by controlling EMTrelated protein expression. Mechanistically, RP11156L14.1 exerted its function as a competing endogenous RNA (ceRNA) and directly interacted with miR548ao3p. The present study also demonstrated that miR548ao3p regulated signal sequence receptor subunit 1 (SSR1) expression by targeting SSR1 3'UTR. Moreover, the xenograft HSCC tumor model revealed that knockdown of RP11156L14.1 markedly suppressed HSCC tumor growth in vivo. In summary, these findings indicated that the lncRNA RP11156L14.1 functions as an oncogene in HSCC by competing with miR548ao3p in regulating SSR1 expression. The RP11156L14.1/miR548ao3p/SSR1 axis could be utilized as a potential novel biomarker and therapeutic target for HSCC.
Assuntos
Proteínas de Ligação ao Cálcio/genética , Neoplasias Hipofaríngeas/genética , Glicoproteínas de Membrana/genética , MicroRNAs/metabolismo , RNA Longo não Codificante/metabolismo , Receptores Citoplasmáticos e Nucleares/genética , Receptores de Peptídeos/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/genética , Adulto , Idoso , Animais , Carcinogênese/genética , Linhagem Celular Tumoral , Movimento Celular/genética , Proliferação de Células/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Humanos , Neoplasias Hipofaríngeas/patologia , Neoplasias Hipofaríngeas/cirurgia , Hipofaringe/patologia , Hipofaringe/cirurgia , Laringectomia , Masculino , Camundongos , Pessoa de Meia-Idade , Prognóstico , RNA Longo não Codificante/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço/patologia , Carcinoma de Células Escamosas de Cabeça e Pescoço/cirurgia , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
The source of seed cells is a critical factor for tissue engineering. The goal of this study was to evaluate the chondrogenesis of Sox9-overexpressing human umbilical cord mesenchymal stem cells (hUCMSCs) seeded onto bone matrix gelatin (BMG)/fibrin hybrid scaffolds both in vitro and in vivo. hUCMSCs were stably transfected with Sox9-expressing plasmid and grown on the three-dimensional BMG/fibrin hybrid scaffold for 8 weeks. Scanning electron microscopy and histochemistry were performed. The hUCMSC-loaded scaffolds were implanted into the subcutaneous layer of immunocompetent rats and chondrogenesis and host immune responses were monitored for 8 weeks. We found that hUCMSCs spread well and proliferated from 2 weeks after culturing. They produced abundant glycosaminoglycans and collagen II. At 8 weeks after implanting into rats, the hUCMSCs on the scaffolds formed cartilage-like tissue and displayed positive staining for toluidine blue, safranin O, Masson's trichrome, and collagen II. No significant changes in serum levels of lgG, lgA, lgM, C3, and C4 were observed after implantation of the hUCMSC-loaded scaffolds. Xenogeneic implantation of Sox9-overexpressing hUCMSCs embedded in the BMG/fibrin scaffolds promotes the formation of cartilage-like tissue without inducing evident host immune response. Therefore, Sox9-overexpressing hUCMSCs represent a promising cell candidate for cartilage tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1150-1155, 2017.
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Cartilagem/metabolismo , Sangue Fetal/metabolismo , Células-Tronco Mesenquimais/metabolismo , Fatores de Transcrição SOX9/biossíntese , Engenharia Tecidual , Alicerces Teciduais/química , Animais , Cartilagem/citologia , Sangue Fetal/citologia , Xenoenxertos , Humanos , Transplante de Células-Tronco Mesenquimais , Células-Tronco Mesenquimais/citologia , Ratos , Ratos Sprague-Dawley , Fatores de TempoRESUMO
OBJECTIVES/HYPOTHESIS: To compare the different effects of bronchoalveolar lavage (BAL) with diverse combinations of lidocaine, epinephrine, and dexamethasone on pediatric patients with an inhaled tracheobronchial foreign body (TFB). STUDY DESIGN: Randomized controlled study. METHODS: Two hundred forty cases of pediatric patients with inhaled TFB were included in this study, and were randomly divided into four groups using three kinds of drugs for BAL, namely 0.9% saline (S) group, 2% lidocaine with diluted epinephrine (LE) group, 2% lidocaine with diluted epinephrine and 0.5% dexamethasone (LED), control group (C) without BAL. The incidences of intraoperative or postoperative complications and recovery periods were compared. Meanwhile, the concentrations of interleukin (IL)-1ß, IL-6, and tumor necrosis factor (TNF)-α in BAL fluids and plasma were evaluated by enzyme-linked immunosorbent assay. RESULTS: The incidences of bronchospasm, hypoxemia, and postoperative fever were significantly lower in the LED group than other groups (P < .001). Fever after the TFB removal procedure appeared later in the LED group than the other groups. The improvement and healing periods in the LE and LED groups were significantly shorter than those in the C and S groups (P < .001). The concentrations of IL-1ß, IL-6, and TNF-α in BAL fluids were significantly higher in the LE and LED groups than those in the S group (P < .001), but those in the plasma of the C and S groups were lower compared with the LE and LED groups (P < .001). CONCLUSIONS: BAL with lidocaine, epinephrine, and dexamethasone could promote recovery for TFB patients and reduce incidences of complications, possibly by regulating release of proinflammatory cytokines. LEVEL OF EVIDENCE: 1b.
Assuntos
Lavagem Broncoalveolar/métodos , Corpos Estranhos/terapia , Anestésicos Locais/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Biomarcadores/análise , Broncodilatadores/uso terapêutico , Broncoscopia , Pré-Escolar , Dexametasona/uso terapêutico , Epinefrina/uso terapêutico , Feminino , Humanos , Incidência , Interleucina-1beta/análise , Interleucina-6/análise , Complicações Intraoperatórias/epidemiologia , Lidocaína/uso terapêutico , Masculino , Complicações Pós-Operatórias/epidemiologia , Fatores de Tempo , Fator de Necrose Tumoral alfa/análiseRESUMO
OBJECTIVE: To investigate the concentrations of pepsin and pepsinogen within the middle ear cavity and determine whether pepsin and pepsinogen affect the prognosis of children with otitis media with effusion (OME). METHODS: All middle-ear lavage fluid from patients with OME undergoing myringotomy (M subgroup) or tympanostomy tube insertion (T subgroup) was collected and pepsin and pepsinogen were detected using enzyme-linked immunosorbent assay. After close follow-up over 2 years, the effects of pepsin and pepsinogen on the prognosis of the patients with OME in the M and T subgroups were analyzed. RESULTS: The average pepsin and pepsinogen concentrations were significantly lower in the M subgroup (n=54; 24.38±16.10mg/mL and 286.49±91.95mg/mL, respectively) than in the T subgroup (n=55; 45.56±16.60mg/mL and 664.92±107.06mg/mL; t=2.484, P=0.018 and t=2.670, P=0.011, respectively). In the M subgroup, the average time to tympanic membrane healing and tympanic pressure restoration to normal was much longer in pepsin(+) patients (17.0±2.0 days and 26.0±2.5 days, respectively) than in pepsin(-) patients (14.0±1.1 days and 22.0±1.0 days; t=3.871, P=0.001 and t=5.734, P=0.000, respectively), and the hearing level of pepsin(+) patients with OME ascended to 13.08±1.19dB, which was much lower than that of pepsin(-) patients (18.29±1.27dB; t=11.001, P=0.000). In the T subgroup, the complication rate including otorrhea and myringosclerosis was much higher in patients with high pepsin concentrations than in those with low pepsin concentrations (P<0.05). Finally, in both subgroups, the recurrence rates of OME in pepsin(+) or patients with high pepsin concentrations (34.6% [9/26] and 28.6% [10/35]) were significantly higher than those in pepsin(-) or low pepsin concentrations (10.7% [3/28] and 5.0% [1/20]; χ(2)=4.456, P=0.035 and χ(2)=4.420, P=0.036). However, pepsinogen had no significant effect on OME prognosis or recurrence. CONCLUSION: Pepsin but not pepsinogen could postpone tympanic membrane healing and pressure restoration in children with OME undergoing myringotomy and increase the incidence of recurrence and complications including otorrhea and myringosclerosis for those undergoing tympanostomy tube insertion. Therefore, pepsin could be considered a poor prognostic factor for OME, further emphasizing the important role of pepsin in OME pathogenesis.
Assuntos
Ventilação da Orelha Média , Otite Média com Derrame/enzimologia , Otite Média com Derrame/cirurgia , Pepsina A/análise , Pepsinogênio A/análise , Membrana Timpânica/cirurgia , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Seguimentos , Audição , Humanos , Masculino , Ventilação da Orelha Média/efeitos adversos , Otite Média com Derrame/fisiopatologia , Pressão , Prognóstico , Recidiva , Membrana Timpânica/fisiologia , CicatrizaçãoRESUMO
OBJECTIVES/HYPOTHESIS: To analyze the relationship between laryngopharyngeal reflux (LPR) represented by pepsin and pepsinogen, and pathogenesis of otitis media with effusion (OME). STUDY DESIGN: Prospective case-control study. METHODS: Children with OME who required adenoidectomy and tympanostomy/tympanostomy tubes placement were enrolled in OME group, whereas children with adenoid hypertrophy (AH) who required adenoidectomy and individuals who required cochlear implantation (CI) were enrolled in AH and CI groups, respectively. Pepsinogen mRNA and protein levels were assessed by real-time fluorescence-based quantitative polymerase chain reaction and immunohistochemistry in adenoid specimens from the OME and AH groups. Pepsin and pepsinogen concentrations were evaluated by enzyme-linked immunosorbent assay in middle ear fluid and plasma from the OME and CI groups. RESULTS: The levels of pepsinogen protein expressed in cytoplasm of epithelial cells and clearance under epithelial cells in adenoid specimens from the OME group were significantly higher than those in the AH group. Furthermore, the concentrations of pepsin and pepsinogen in the OME group were 51.93±11.58 ng/mL and 728±342.6 ng/mL, respectively, which were significantly higher than those in the CI group (P<.001). In addition, the concentrations of pepsin in dry ears were significantly lower than those in serous and mucus ears in the OME group (F=22.77, P<.001).Finally, the concentration of pepsinogen in middle ear effusion was positively correlated with the expression intensity of pepsinogen protein in cytoplasm of epithelial cells (r=0.73, P<.05) in the OME group. CONCLUSIONS: Pepsin and pepsinogen in middle ear effusion are probably caused by LPR and may be involved in the pathogenesis of OME. LEVEL OF EVIDENCE: 3b.
Assuntos
Regulação da Expressão Gênica , Refluxo Laringofaríngeo/complicações , Otite Média com Derrame/etiologia , Pepsina A/genética , Pepsinogênio A/genética , Tonsila Faríngea/química , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Monitoramento do pH Esofágico , Feminino , Seguimentos , Humanos , Imuno-Histoquímica , Refluxo Laringofaríngeo/genética , Refluxo Laringofaríngeo/metabolismo , Masculino , Otite Média com Derrame/genética , Otite Média com Derrame/metabolismo , Pepsina A/biossíntese , Pepsinogênio A/biossíntese , Estudos Prospectivos , RNA Mensageiro/análise , RNA Mensageiro/genética , Reação em Cadeia da Polimerase em Tempo RealRESUMO
Our previous studies have showed that chemokine receptor 4 (CXCR4) was over-expressed in laryngeal squamous cell carcinoma (LSCC). However, the mechanism underlying aberrant CXCR4 expression remains unclear. To investigate the roles played by miRNAs in CXCR4 over-expression in LSCC, putative miR-139 was predicted through computational algorithms, including TargetScan, PicTar and miRBase, and luciferase reporter assay was explored to confirm that whether CXCR4 was directly regulated by miR-139. Then, quantitative real-time PCR, immunohistochemistry and in situ hybridization methods were employed to detect the expression of miR-139 and CXCR4 in primary LSCC tissues, normal adjacent mucosal tissues and metastatic lesions derived from 40 LSCC patients in the Second Hospital, Xi'An JiaoTong University. Finally, gain- and loss-of-function assays were adopted to explore the effects of miR-139 and CXCR4 on proliferation, invasion and metastasis of the human LSCC cell line Hep-2 in vitro and in vivo. Our results showed that miR-139 dampened CXCR4 expression, and CXCR4 was directly targeted by miR-139. Additionally, the expression of miR-139 was reduced in alignment with the progression of primary to metastatic LSCC. Moreover, an inverse correlation was observed between miR-139 and CXCR4 protein levels in LSCC specimens. Functional analyses demonstrated that ectopic expression of miR-139 inhibited cell proliferation, migration and metastasis of Hep-2 cells in vitro and in vivo. Similar to the observations seen in restoring miR-139 expression, dampening of CXCR4 expression inhibited cell growth, migration and invasion, whereas miR-139 over-expression reversed the pro-metastatic effect of CXCR4. Taken together, we conclude that miR-139 targets CXCR4 and inhibits proliferation and metastasis of LSCC.
Assuntos
Carcinoma de Células Escamosas/metabolismo , Regulação Neoplásica da Expressão Gênica , Neoplasias Laríngeas/metabolismo , MicroRNAs/metabolismo , Receptores CXCR4/metabolismo , Animais , Linhagem Celular Tumoral , Proliferação de Células , Perfilação da Expressão Gênica , Células HEK293 , Humanos , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Metástase Neoplásica , Transplante de NeoplasiasRESUMO
Nasopharyngeal carcinoma (NPC) is uncommon worldwide but often highly invasive in late stages. Due to its special location and lack of specific symptoms, NPC is hardly detected in regular medical examination at the beginning. Development of sensitive and specific biomarkers should help to save lives against this type of disease. In the present report, we investigated the value of plasma miRNAs for diagnosis and prognosis of NPC. Using candidate approach, we selected 21 miRNAs from literature to compare their expression levels in the plasma of NPC patients and controls. As a result, 5 miRNAs showed diagnostic potentials (P<0.01). Among them, miR-16, -21, -24, and -155 had increased levels in NPC patients, whereas the level of miR-378 was decreased. There was a negative correlation between plasma miRNA expression and cancer progression, where miR-21 was statistically significant in T and N staging and miR-16 and 24 were significant in N staging only. Combination of miR-16, -21, -24, -155, and -378 gives 87.7% of sensitivity and 82.0% of specificity for NPC diagnosis. Without miR-16, combination of the rest 4 miRNAs gives the same sensitivity but a slightly reduced specificity. After treatment, all 5 miRNAs were somewhat back to normal levels in patients without cancer recurrence but the prognostic value was not statistically significant. In conclusion, plasma miRNA expression is a useful biomarker for NPC diagnosis but not for its prognosis. More importantly, it is simple, effective, and non-invasive. Combination of several plasma miRNAs can increase both NPC diagnostic sensitivity and specificity.
Assuntos
Biomarcadores Tumorais/sangue , MicroRNAs/sangue , Neoplasias Nasofaríngeas/diagnóstico , Adolescente , Adulto , Idoso , Biomarcadores Tumorais/genética , Carcinoma , Feminino , Humanos , Masculino , MicroRNAs/genética , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/patologia , Prognóstico , Adulto JovemRESUMO
OBJECTIVE: To investigate the effects of the enhancer of zeste homolog 2 (EZH2) gene on cell growth and invasion of the nasopharyngeal carcinoma (NPC). METHODS: Recombinant lentivirus vector for shRNA delivery of EZH2 was constructed and transfected into 293FT cells. After collecting the viral particles, the NPC cell line 5-8F cells were transfected. The effects of EZH2 silence on cell proliferation and cell cycle were detected using MTT assay, plate colony formation assay and flow cytometry. The migration and invasion of 5-8F cells were determined by wound healing assay and matrigel invasion assay, respectively. The expressions of EZH2 and epithelial-mesenchymal transition (EMT)-related markers at mRNA and protein levels were examined by real-time PCR and Western blot respectively. RESULTS: The expressions of EZH2 mRNA and protein in the transfected 5-8F cells were obviously reduced. MTT assay showed that EZH2 downregulation significantly inhibited the growth of 5-8F/shEZH2 cells (P < 0.001). Colony formation rate (84.44%) of 5-8F/shEZH2 cells was lower than control (31.56%, P = 0.001). Cell cycle analysis showed that most 5-8F/shEZH2 cells were arrested in G0/G1 phase, with a very low ratio of cells in S phase. Wound healing assay indicated that the migration ability of cells silencing EZH2 decreased significantly, and the 48-hour relative migration distance of 5-8F/ShEZH2 cells and control cells was 0.58 ± 0.05, and 0.81 ± 0.02, respectively (P < 0.000). Matrigel invasion assay, showed the invasive capacity of cells silencing EZH2 was significantly inhibited, with less penetrating cells (72.23 ± 4.08) compared to control (150.95 ± 16.27), P < 0.000. The mRNA expressions of epithelial markers E-cadherin and Keratin 18 in the cells silencing EZH2 increased by 177% and 158% respectively, and the mRNA expressions of mesenchymal markers ß-catenin and N-cadherin decreased by 18.04% and 41.18% respectively. Similar results also were obtained with Western blot analysis. CONCLUSION: EZH2 significantly enhanced the proliferation and invasion of nasopharyngeal carcinoma cells in vitro, which might be mediated by inducing EMT.
Assuntos
Proliferação de Células , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/patologia , Complexo Repressor Polycomb 2/genética , Carcinoma , Linhagem Celular Tumoral , Proteína Potenciadora do Homólogo 2 de Zeste , Transição Epitelial-Mesenquimal , Regulação Neoplásica da Expressão Gênica , Humanos , Carcinoma Nasofaríngeo , Invasividade NeoplásicaRESUMO
OBJECTIVE: To study the effect of Epstein-Barr virus nuclear antigen 1 (EBNA1) on cell proliferation and cell cycle in nasopharyngeal carcinoma (NPC) cells. METHODS: Recombinant lentivirus that encoded EBNA1 short hairpin RNA (shRNA) was prepared. The C666-EBNA1 (CE) cells were transduced with lentivirus and selected by fluorescence activated cell sorting (FACS) to repress EBNA1 expression. The protein expression levels of EBNA1 were examined by Western blot. The effect of EBNA1 silence on cell proliferation was analyzed by MTT assay and cell growth assay, respectively. Cell cycle was assessed by flow cytometry. The mRNA and protein levels of cell cycle regulators were examined by real-time PCR and Western blot. RESULTS: Recombinant lentivirus encoded EBNA1 shRNA was successfully constructed. The EBNA1 expression in CE cells was significantly reduced by lentivirus-mediated RNA interference. The results of cell counting and MTT assay showed that EBNA1 down-regulation significantly inhibited cell growth in CE-shRNA EBNA1 cells (P < 0.05). Compared with the control group, the percentage of cells in G0-G1 phase was increased from (62.43 ± 6.62)% to (89.66 ± 0.64)% (t = -7.091, P = 0.002), and that in S phase was decreased from (34.93 ± 7.36)% to (7.82 ± 2.44)% (t = 6.095, P = 0.004). The mRNA expressions of c-myc, CDK4, CDK6 and pRb were decreased by 65.60%, 34.06%, 41.05% and 55.29% respectively with the similar results in protein expression levels. CONCLUSIONS: Suppression of EBNA1 may inhibit the growth of nasopharyngeal carcinoma cells in vitro and induce a G1-phase cell cycle arrest, which might be mediated by down-regulation of c-myc, CDK4, CDK6 and pRb.
Assuntos
Ciclo Celular , Proliferação de Células , Antígenos Nucleares do Vírus Epstein-Barr/metabolismo , Neoplasias Nasofaríngeas/metabolismo , Carcinoma , Linhagem Celular Tumoral , Antígenos Nucleares do Vírus Epstein-Barr/genética , Regulação Neoplásica da Expressão Gênica , Vetores Genéticos , Humanos , Lentivirus , Carcinoma Nasofaríngeo , Neoplasias Nasofaríngeas/genética , Neoplasias Nasofaríngeas/patologia , Interferência de RNA , RNA Mensageiro/genética , RNA Interferente Pequeno/genética , Transdução GenéticaRESUMO
OBJECTIVE: To explore the role of CXCR4/SDF-1alpha axis in organ-specific metastasis of nasopharyngeal carcinoma (NPC) by assessment of CXCR4 expression in NPC cells and SDF-1alpha expression in distant target organs of NPC. METHODS: Thirty patients with NPC and fifteen normal subjects were recruited in this study. The expressions of CXCR4 in NPC and normal cases were identified by RT-PCR and immunohistochemistry (IHC), then the relationship between CXCR4 expression and clinicopathological factors was analyzed. IHC was also used to analyze the SDF-1alpha protein expression in normal cervical lymph nodes (including normal superior and inferior deep cervical lymph nodes), bone marrow, lung, liver, kidney and colon tissues of NPC patients (5 cases/each group). RESULTS: The relative expression level of CXCR4 mRNA in NPC (0.71 +/- 0.22) was significantly higher than that of normal nasopharynx tissues (0.14 +/- 0.07, F = 27.94, P < 0.05). The relative expression level of CXCR4 protein in NPC (1.58 +/- 0.59) was significantly higher than that of normal nasopharynx tissues (0.51 +/- 0.22, F = 17.75, P < 0.05). The high expression levels of CXCR4 mRNA and protein in NPC were closely related to clinical stage, cervical lymph node metastasis and cancer cell differentiation (P < 0.05). SDF-1alpha protein was strongly expressed in normal superior deep cervical lymph nodes, bone marrow, lung and liver (2.35 +/- 0.67), but absent or very poor expression in inferior deep cervical lymph nodes, kidney and colon tissues (0.68 +/- 0.23), and the differences between them were statistically significant (t = 10.13, P < 0.01). CONCLUSION: CXCR4 is closely correlated to metastasis of nasopharyngeal carcinoma. CXCR4/SDF-1alpha axis may play an important role in organ-specific metastasis of NPC.