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BACKGROUND: Tumor necrosis factor-associated protein 1 (TRAP1) is a heat-shock protein 90-related mitochondrial chaperone. Accumulative evidence has demonstrated that TRAP1 overexpression is closely related to carcinogenesis. However, the exact function and mechanism of TRAP1 in the occurrence of laryngeal carcinoma remains unclear. OBJECTIVE: To investigate whether RNA interference can inhibit TRAP1 overexpression and to explore its effects on growth and apoptosis of CD133+CD44+ laryngeal carcinoma stem cells. METHODS: CD133+CD44+ laryngeal carcinoma stem cells were sorted from human laryngeal carcinoma Hep-2 cellsusing immunomagnetic beads. The shRNA sequence of TRAP1 was designed and synthesized and CD133+CD44+ laryngeal carcinoma stem cells were transfected with LipofectamineTM 2000. Cell counting kit-8 assay, colony formation assay and flow cytometry were used to investigate the effects of interference of TRAP1 expression on growth and apoptosis of CD133+CD44+ laryngeal carcinoma stem cells. Spectrophotometric method was used to detect the activity of caspase-3, -8 and -9. RESULTS AND CONCLUSION: TRAP1 mRNA and protein expression levels were significantly decreased in TRAP1 shRNA-transfected CD133+CD44+ laryngeal carcinoma stem cells (P < 0.01). Compared with the blank control and negative control groups, the growth and colony formation of CD133+CD44+ laryngeal carcinoma stem cells were significantly inhibited in the TRAP1 shRNA-transfected group (P < 0.05). Apoptosis of CD133+CD44+ laryngeal carcinoma stem cells was significantly inhibited in the TRAP1 shRNA-transfected group as compared with the blank control and negative control groups (P < 0.05). TRAP1 shRNA-mediated cell apoptosis was associated with the activation of caspase-3, -8 and -9. These results suggest that RNA interference targeting inhibition of TRAP1 suppresses cell growth but promotes apoptosis in CD133+CD44+ aryngeal carcinoma stem cells. TRAP1 is likely to be a gene target for treatment of laryngeal carcinoma.
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BACKGROUND:Studies have indicated that the abnormal expression of tumor necrosis factorreceptor-associated protein 1 (TRAP1) is closely related to the occurrence and development of a variety of tumors. Therefore, targeted inhibition of TRAP1 expression has become an important target for the treatment or intervention of tumor growth. OBJECTIVE:To explore the effect of the TRAP1 gene silencing on the proliferation and apoptosis of human laryngeal squamous cel carcinoma stem cel s. METHODS:CD24-CD44-human laryngeal squamous cel carcinoma stem cel s were isolated by flow cytometry. Interfering RNA (siRNA) sequences for smal molecule TRAP1 gene was designed and transferred into human laryngeal cancer stem cel s by LipofectamineTM 2000. Flow cytometry, MTT assay, cel clone formation assay and TUNEL apoptosis assay were used to evaluate the effect of silencing TRAP1 gene on the proliferation and apoptosis of CD24-CD44+laryngeal cancer stem cel s. RESULTS AND CONCLUSION:Compared with CD24+CD44-cel s, CD24-CD44+cel s upregulated OCT4, SOX2, NANOG and TRAP1 expression levels (P<0.05). However, the expression of TRAP1 protein in human laryngeal squamous cel carcinoma was significantly decreased after RNA interference (P<0.05). The growth rate of TRAP1 gene silenced human laryngeal squamous cel carcinoma was significantly reduced (P<0.05), the cel arrest was in the G0/G1 phase, the number of cel s in the S phase was decreased (P<0.05), and there was no significant change in the M phase. TRAP1 gene silencing significantly inhibited the proliferation of human laryngeal squamous cel carcinoma stem cel s (P<0.05). Compared to the non-transfected cel s, the TRAP1 gene silencing significantly reduced the clone formation ability of transfected human laryngeal squamous cel carcinoma stem cel s (P<0.05), and TRAP1 gene silenced-human laryngeal squamous cel carcinoma stem cel s were more easy to trigger apoptosis by upregulating BAD and BAX expression levels (P<0.05). Overal , our experimental results indicate that the specific interference of TRAP1 gene expression could inhibit the proliferation and promote apoptosis of human laryngeal squamous cel carcinoma stem cel s.
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BACKGROUND:Several studies have demonstrated that JAK2 kinase plays an important role in myocardial protection by ischemic preconditioning and medicine preconditioning. OBJECTIVE:To investigate the effects of exercise preconditioning on JAK2 mRNA and protein expression in the rat heart and the underlying mechanism of action. METHODS:SD rats were randomly divided into control, exhaustive, exercise preconditioning, and exercise preconditioning + AG490 groups. Rats in the control and exhaustive groups were fed routinely for 3 days. Rats in the exercise preconditioning and exercise preconditioning + AG490 groups were subjected to intermittent treadmil exercise for 3 successive days to establish exercise preconditioning animal models. Rats in the exercise preconditioning + AG490 group were intraperitonealy injected with JAK2 inhibitor AG490 at 10 minutes before exercise preconditioning. After 3 days, rats in the exhaustive, exercise preconditioning, and exercise preconditioning + AG490 groups were subjected to exhaustive treadmil exercise. Serum levels of lactate dehydrogenase and creatine kinase isoenzyme were determined. JAK2 mRNA and protein expression in the rat myocardial tissue was detected by real-time fluorescent quantitative PCR and immunohistochemistry.RESULTS AND CONCLUSION:Compared with the control group, serum levels of actate dehydrogenase and creatine kinase isoenzyme were increased, and JAK2 mRNA and protein expression in the myocardial tissue was significantly increased in the exhaustive group (P < 0.05). Compared with the exhaustive group, serum levels of actate dehydrogenase and creatine kinase isoenzyme were decreased (P < 0.05), while JAK2 mRNA and protein expression in the myocardial tissue was increased in the exercise preconditioning group (P < 0.05). Compared with the exercise preconditioning group, serum levels of actate dehydrogenase and creatine kinase isoenzyme were increased (P < 0.05), while JAK2 mRNA and protein expression in the myocardial tissue was decreased in the exercise preconditioning + AG490 group (P < 0.05). The results confirm that exercise preconditioning plays its cardioprotective role through activating JAK/STAT signaling pathway, up-regulating the expression of JAK2 mRNA and protein in the cardiac tissue and aleviating myocardial ischemia injury.
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Objective To investigate the effect of lysyl oxidase (LOX) gene expression on proliferation, invasion and radiotherapy sensitivity in laryngeal cancer Hep-2 cells. Methods Hep-2 cells were divided into control group (normal cultured), negative control group (transfection reagent) and transfection group (LOX siRNA transfected). The expressions of LOX, Ki-67, PCNA, MMP-2 and MMP-9 mRNA were detected by real time-PCR and Western blot methods. The cell sur-vival rate and apoptosis irradiated by different doses of X-rays (0, 3, 6, 9, 12, 15 and 18 Gy) were detected by MTT and flow cytometry (FCM). Results The expression levels of LOX, Ki-67, PCNA, MMP-2 and MMP-9 protein and mRNA were sig-nificantly lower in Hep-2 cells after transfection than those of control group and negative control group (P<0.05). The cell survival rate was significantly inhibited 24 hours after irradiation (12, 15 and 18 Gy) in a dose-dependent manner (P<0.05). The apoptotic rate of transfection cells combined with radiotherapy was significantly higher than that of control and the nega-tive control groups (%:79.11 ± 1.26 vs 5.01 ± 1.02, 4.95 ± 1.12, 43.21 ± 2.1,P<0.05). Conclusion The expression of LOX can be down-regulated after LOX siRNA transfection, inhibiting proliferation and enhancing radiosensitivity, which may be related to the down-regulation of Ki-67, PCNA, MMP-2 and MMP-9 protein expression.
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Objective To observe changes of erythropoietin (EPO) and rheology in moderate and severe obstructive sleep apnea hypopnea syndrome (OSAHS) patients after the noninvasive positive pressure ventilation (NPPV) treatment. Methods Healthy adults were selected as control group (n=40) while moderate to severe OSAHS patients were selected as OSAHS group. OSAHS group was underwent NPPV treatment then, Levels of sleep apnea hypopnea index (AHI), sleep mean minimum oxygen saturation (LSaO2) and serum erythropoietin (EPO) were assessed, routine blood test and hemodynam-ic indexes were also checked before treatment and 1 and 3 months after treatment in both groups. Results In both groups serum EPO, blood, blood rheology indexes, AHI, LSaO2 were analysised at each time point by ANOVA repeated measures, all of which show significant different between groups and between each time points within the same group. Indexes in OSAHS group improved with prolonged treatment , but they are in the normal range in the control. Conclusion OSAHS pa-tients with NPPV therapy can significantly relieve hypoxia, reduce serum EPO level and blood viscosity. So NPPV has impor-tant clinical significance in prevention and treatment of OSAHS.
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OBJECTIVE@#The utility of virtual endoscopy is compared to fiberoptic endoscopy and was also investigated with respect to accuracy of diagnosis and reproduction of images in patients with obstructive sleep apnea hypopnea syndrome (OSAHS).@*METHOD@#Twenty-one patients with OSAHS were examined by helical spiral CT axial images and fiberoptic endoscopy. The helical spiral CT axial data was reconstructed using a VE software program. The results were compared to the fiberoptic endoscopic findings done by the otolaryngologists. All the patients were examined both in the sleeping and nonsleeping status. The dimensions of soft palate, uvula, lingua and epiglottis region were evaluated.@*RESULT@#The results both in the virtual endoscopy evaluation and fiberoptic endoscopy was statistically significant difference in all of region. In the palate region, there was statistically significant difference in the left-and-right dimension, but no difference in the fore-and-aft dimension both in the sleeping and nonsleeping status.@*CONCLUSION@#The dimensions of upper airway were more difference between in the sleeping and nonsleeping status. The change of dimension was more in the left-and-right; the fiberoptic endoscopy has more diagnostic significance in evaluation of dynamic movement of the upper airway; virtual endoscopy evaluation of the upper airway was accurate in assessing stenosis width and length; virtual endoscopy added more information about anatomic structure and pathological change of the upper airway.
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Adult , Female , Humans , Male , Middle Aged , Endoscopy , Methods , Laryngoscopy , Sleep Apnea, Obstructive , Diagnosis , Diagnostic Imaging , Tomography, Spiral Computed , Tomography, X-Ray ComputedABSTRACT
OBJECTIVE On the basis of the different shapes of upper airway(UA)obstruction and collapse characteristic of obstructive sleep apnea hypopnea syndrome(OSAHS), the effect of modified uvulopalatopharyngoplasty(UPPP)combined with coblation in OSAHS operations on patients with AHI≥20,which could be done selectively to patients with various or multiple UA obstructive sites, was explored. METHODS 30 patients of OSAHS were diagnosed and analysed by polysomnography(PSG)to have AHIs≥20. Based on the shapes of their UAs classified according to the degree of the shortened lateral and radial vectors,the UPPP was modified in two ways: either the lateral vector or the radial vector was amplified, or both could be amplified with coblation. All patients were analyzed by questionnaires(PSG)Muller's maneuver and oral cavity measurement pre-and 6 months post-operation. RESULTS 86.7 % of the patients showed a decrease of at least 25 % in AHI . Velopalatal insufficiency never occurred. CONCLUSION The modified-UPPP combined with coblation can be used selectively on patients based on their UA shapes.