Publications (18)23.04 Total impact
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Article: [Role of ABCB1 and ABCG2 in the multidrug resistance of hypopharyngeal carcinoma FaDu cell line].
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ABSTRACT: To investigate the expression of multidrug resistance gene ABCB1 and ABCG2 in FaDu cells (human hypopharyngeal carcinoma cell line) and the multidrug resistance (MDR) cell lines FaDu/T transformed from FaDu cells by taxol and underlying mechanisms of MDR. The multidrug resistance sensitivities of FaDu and FaDu/T to cisplatin (DDP), 5-fluorouracil (5-FU), doxorubicin (Dox), and vincristine (VCR) were examined by methyl-thiazolyl-tetrazolium (MTT) assay. The mRNA and protein expressions of multidrug resistance genes ABCB1 and ABCG2 were analysed with RT-PCR, Western blot and laser confocal microscopy. JNK signal proteins were detected through Western blot. The multidrug resistance of FaDu/T cells to Taxol, DDP, 5-FU, ADM and VCR was more than that of FaDu cells. The expression of ABCB1 in FaDu/T cells was significantly higher than that in FaDu cells (t = 22.42, P < 0.05), but the expression of ABCG2 in FaDu/T cells was significantly lower than that in FaDu cells (t = 10.06, P < 0.05). JNK signal was inhibited in FaDu or FaDu/T cells and the inhibited JNK was reactivated by taxol or anisomycin (an activator for MAPK signal transduction pathways). Anisomycin down-regulated the expression of ABCB1 (F = 33.72, P < 0.05) and up-regulated the expression of ABCG2 (F = 220.16, P < 0.05) in FaDu/T cells, but not in FaDu/T cells pretreated by JNK inhibitor SP600125 (P > 0.05). The overexpression of ABCB1 and the down-regulation of ABCG2 in FaDu/T cells were the main features of MDR in hypopharyngeal carcinomas, in which JNK signal transduction pathways could play an important role.Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery 04/2012; 47(4):305-10. -
Article: Nimesulide inhibited the growth of hypopharyngeal carcinoma cells via suppressing Survivin expression.
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ABSTRACT: The objective of this study was to evaluate the efficacy of Nimesulide, a selective cyclooxygenase-2 (COX-2) inhibitor, on the growth of hypopharyngeal carcinoma cells (FaDu) in vitro, and investigate its potential mechanism. After FaDu cells were treated with graded concentrations of Nimesulide for divergent time, sensitivity of cells to drug treatment was analyzed by MTT assay. Morphological changes of FaDu cells in the presence of Nimesulide were observed by acridine orange cytochemistry staining. Proliferating cells were detected using the 5-Bromo-2'-deoxy-uridine (BrdU) incorporation assay. Following cells were subjected to Nimesulide (500 μmol/l) for 6 h, 12 h and 24 h, the percentage of apoptosis was examined by flow cytometry. We detected COX-2 and Survivin expression change by RT-PCR and Western blot, and analyzed the correlation of them with the growth of FaDu cells. Additionally, we also analyzed Caspase-3, Bcl-2 and Bax expressions as markers to investigate the related pathway of Nimesulide-indued apoptosis. Compared with the control group, the viabilities rates were decreased by Nimesulide in time- and dose-dependent manners, typical morphological changes of apoptotic cells were observed in the Nimesulide-treatment groups, Nimesulide could suppress the proliferation of FaDu cells significantly. The percentage of apoptosis in FaDu cells were markedly increased after Nimesulide-treatment for 6 h, 12 h and 24 h. Nimesulide down-regulated the Survivin and COX-2 expressions at mRNA and protein levels in FaDu cells. Additional analyses indicated that Bcl-2 expression was significantly decreased and the expressions of Caspase-3 as well as Bax were increased at both mRNA and protein levels. Based on the induction of apoptosis and suppression of proliferation, Nimesulide could inhibit the growth of FaDu cells. Furthermore, the suppression of Survivin expression may play an important role in Nimesulide-induced growth inhibition. Nimesulide could act as an effective therapeutic agent for hypopharyngeal carcinoma therapy.Head & Neck Oncology 03/2012; 4:7. · 3.13 Impact Factor -
Article: TWIST expression in hypopharyngeal cancer and the mechanism of TWIST-induced promotion of metastasis.
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ABSTRACT: The transcription factor TWIST is an important factor in regulating epithelial-mesenchymal transition (EMT) and tumor metastasis. To explore the functions of TWIST in hypopharyngeal cancer, we investigated if overexpression of TWIST has an effect on FaDu cell morphology, and if alteration of TWIST has an effect on E-cadherin, N-cadherin, c-fos, MMP-9, as well as in cell migration, and the invasion ability of FaDu cells. Moreover, we also studied the relationship between TWIST overexpression and clinicopathological characteristics in hypopharyngeal cancer tissue samples by immunohistochemical assays. The results showed that overexpression of TWIST-induced morphological changes, such as occurrence of EMT. TWIST overexpression also increased cell migration and invasion ability, accompanied by an alteration of E-cadherin, N-cadherin, c-fos and MMP-9 expression. Furthermore, immunohistochemical assays showed that TWIST overexpression was related with tumor differentiation (P=0.038), tumor size (P=0.048) and lymph node metastasis (P=0.044). The data presented reveal that overexpression of TWIST plays a significant role in the metastasis of hypopharyngeal tumors, and alteration of TWIST has an effect on the EMT, c-fos and MMP-9 expression in FaDu cells. We conclude that TWIST promotes hypopharyngeal carcinoma metastasis, and the TWIST/c-fos/MMP-9 signaling pathway may play an important role in the metastasis of FaDu cells.Oncology Reports 02/2012; 27(2):416-22. · 1.84 Impact Factor -
Article: L-cysteine attenuates peroxynitrite-elicited cytotoxicity to spiral ganglion neurons: possible relation to hearing loss.
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ABSTRACT: The purpose of this work was to investigate whether L-cysteine was able to protect spiral ganglion neurons (SGNs) against peroxynitrite (ONOO(-))-elicited toxicity. The rat SGNs were isolated and cultured in this work. Cell viability was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The morphological changes were examined under inverted phase contrast microscope. Cells underwent apoptosis were determined by TdT-mediated dUTP nick end labeling (TUNEL) assay. Intracellular glutathione (GSH) content, superoxide dismutase (SOD) activity and malonaldehyde (MDA) level were detected by biochemical methods. Laser scanning confocal microscope was employed to analyze cytosolic Ca(2+) concentration. Results showed that ONOO(-) reduced the cell viability of SGNs in a time- and dose-dependent manner. ONOO(-)-triggered cell damage was further confirmed via apoptotic pathway rather than necrosis. Pretreatment with L-cysteine (5 mM) for 12 hours could almost completely rescue SGNs from ONOO(-)-induced damage. The decrease in intracellular GSH content and SOD activity, as well as the increase in MDA level induced by ONOO(-) were correspondingly antagonized by the administration of L-cysteine. Furthermore, L-cysteine can significantly inhibit elevation of Ca(2+) concentration induced by ONOO(-). Our findings indicate that L-cysteine protects SGNs from ONOO(-)-induced damage via enhancing the antioxidative activity and, suppressing the lipid peroxidation as well as the release of cytosolic Ca(2+), thereby indicating that oxidation resistance was useful to prevent audiological diseases initiated by oxidative stress.Neurological Research 11/2011; 33(9):935-41. · 1.52 Impact Factor -
Article: FaDu cell characteristics induced by multidrug resistance.
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ABSTRACT: The major obstacle to tumor chemotherapy is drug resistance. In the present study, we investigated the characteristics of FaDu cells (a hypopharyngeal carcinoma cell line) with multidrug resistance (MDR) induced by Taxol. The resistant cell line, FaDu/T, was grown by exposing normal FaDu cells to escalating concentrations of Taxol stepwise for over 12 months. The multidrug resistant sensitivities of the FaDu/T cells to cisplatin (DDP), 5-fluorouracil (5-FU), doxorubicin (Dox) and vincristine (VCR) were investigated by methyl-thiazolyl-tetrazolium (MTT) assay. Cell apoptosis was measured by acridine orange and Hoechst 33342/propidium iodide double staining. Cell cycle distribution and the cell apoptosis index were quantified using flow cytometry. Reverse transcription-polymerase chain reaction (RT-PCR) was conducted to determine the mRNA levels of the MDR-related genes MDR1/ABCB1 and BCRP/ABCG2. Western blotting was used to assay the expression of MDR1/ABCB1 and BCRP/ABCG2 and the apoptosis-related proteins caspase-3, Bcl-2 and Bax. Compared with the FaDu cells, the drug resistance of FaDu/T cells to DDP, 5-FU, Dox and VCR was increased 8.99-, 21.96-, 31.42- and 10.00-fold, respectively. The percentages of FaDu/T cells in the G0/G1 and G2/M phases were increased while the cell percentage in the S phase decreased as compared with the percentages of FaDu cells. The anti-apoptotic ability increased prominently, as the index of apoptosis decreased. Furthermore, caspase-3, Bcl-2 and Bax expression was altered accordingly to resist apoptosis in the FaDu/T cells. MDR1/ABCB1 expression increased significantly at both the mRNA and protein levels, while BCRP/ABCG2 expression appeared to inversely affected, i.e. decreased, in a concentration-dependent manner. These -findings may provide theoretical support for the prevention of MDR in clinical cancer chemotherapy.Oncology Reports 08/2011; 26(5):1189-95. · 1.84 Impact Factor -
Article: Down-regulation of TWIST decreases migration and invasion of laryngeal carcinoma Hep-2 cells by regulating the E-cadherin, N-cadherin expression.
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ABSTRACT: The transcription factor TWIST is an important factor in regulation of the epithelial-mesenchymal transition (EMT), which represents the primary stages during the metastasis of tumors. To identify the role of TWIST in the regulation of metastasis in laryngeal carcinoma Hep-2 cells, we investigated whether the alteration of TWIST has an effect on the Hep-2 cells morphology and whether the alteration of TWIST has an effect on the expression of E-cadherin, N-cadherin as well as the ability of cell motion, migration, and invasion. Morphological changes of Hep-2 cells that were transfected a mircoRNA against TWIST vector were observed by the reserved microscope. Reverse transcription-polymerase chain reaction was performed in order to examine the mRNA expression of TWIST, E-cadherin, and N-cadherin. Western blotting was performed to examine the protein expression of TWIST, E-cadherin, and N-cadherin. Cell motion ability was examined by Scratch-wound assay. Transwell(™) chamber assays were used to determine cell migration and invasion. Transfecting a mircoRNA down-regulated TWIST expression at mRNA and protein levels. Down-regulation of TWIST expression induced morphological changes, such as the inversion of the EMT. Moreover, down-regulation of TWIST expression up-regulated E-cadherin and down-regulated N-cadherin expressions at mRNA and protein levels, respectively. Furthermore, we confirmed that down-regulation of TWIST expression decreased the motion, invasion, and migration ability of the Hep-2 cells. Down-regulation of TWIST expression decreases migration and invasion of laryngeal carcinoma Hep-2 cells by regulation of the E-cadherin, N-cadherin expression.Journal of Cancer Research and Clinical Oncology 08/2011; 137(10):1487-93. · 2.56 Impact Factor -
Article: [Association between lymphangiogenesis and clinicopathological features and prognosis in laryngeal carcinoma].
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ABSTRACT: To detect lymphangiogenesis by labeling the lymphatic endothelial marker, lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), and study the prognostic relevance of lymphangiogenesis in laryngeal squamous carcinoma. Clinical files and specimens of 78 patients with histologically diagnosed laryngeal carcinoma were stained with LYVE-1 as a specific lymphatic endothelial marker. The lymphatic vessel density (LVD) was measured, and the correlation between LVD and clinicopathological features of the tumor cases was analyzed. The mean LVD in laryngeal carcinoma (13.24 ± 5.09) was significantly higher than that in adult laryngeal papilloma (5.54 ± 3.15) and squamous dysplasia (6.76 ± 4.45, P < 0.05). The LVD of poorly differentiated tumors (15.74 ± 5.24) was significantly higher than that in the moderately differentiated tumors (13.84 ± 6.20), and the LVD in the moderately differentiated tumors was significantly higher than that in the well-differentiated tumors (11.68 ± 6.34). The LVD in stage 0 to stage II group (10.66 ± 5.70) was significantly lower than that in the stage III to IV group (17.01 ± 6.35). The lymph node metastasis group (17.25 ± 7.37) was significantly higher than non-lymph node metastasis group (8.60 ± 5.23, P < 0.05). There was no significant association between LVD and age, sex, primary site and distant metastasis. The overall survival in the patients with a LVD higher than the mean value was 33.5 month, and that of cases with a LVD lower than the mean value was 81.6 month (P < 0.05). The multivariate survival analysis showed that the clinical stage and LVD were independent prognostic factors of laryngeal cancer. The LYVE-1 staining histochemistry demonstrates that the lymphangiogenesis occurrs mainly at the edge of the tumors, and lymphangiogenesis plays an important role in the carcinogenesis, cancer progression and lymph node metastasis in laryngeal cancer. LVD may be an independent indicator of poor prognosis of laryngeal cancer.Zhonghua zhong liu za zhi [Chinese journal of oncology] 06/2011; 33(6):461-4. -
Article: Twist modulates lymphangiogenesis and correlates with lymph node metastasis in supraglottic carcinoma.
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ABSTRACT: Twist is a highly conserved epithelial-mesenchymal transcription factor that has been reported to be a key factor in tumor malignancy, including lymph node metastasis. It represents the major step of dissemination and serves as a chief prognostic indicator of disease progression. However, the mechanism by which Twist regulates lymph node metastasis remains incompletely understood. Studies on the mechanism of metastasis are thus required for determining appropriate therapeutic strategies. Immunohistochemistry for lymphatic vessel endothelial receptor 1 (LYVE-1), Ki-67, Twist, vascular endothelial growth factor C (VEGF-C), and vascular endothelial growth factor receptor 3 (VEGFR-3) was performed to detect lymphatic vessel density (LVD), cell proliferation levels and the expressions of Twist, VEGF-C, and VEGFR-3 were determined from 66 primary supraglottic carcinoma tissue samples from 36 patients with lymph node metastasis (pathological N+, pN+) and 30 patients without metastasis (pathological N0, pN0). Western blotting analysis of the proteins in pN+ and pN0 primary tumors was used to characterize the expressions of Twist, VEGF-C, and VEGFR-3 further. The LVD was 22.4 ± 10.3 in pN+ patients and 6.8 ± 4.1 in pN0 ones. For Ki-67, the number of proliferous cells in pN+ patients was greater than that in pN0 ones. Both, however, were associated with their clinical nodal stages. In pN+ patients, Twist, VEGF-C, and VEGFR-3 expressions were 86.11% (31/36), 80.56% (29/36), and 58.33% (21/36), respectively. These values were higher than those found for pN0 patients (i.e., 13/30, 11/30, and 7/30, respectively) (P < 0.05). Among the samples with Twist expression, 88.64% were VEGF-C-positive and 59.09% were VEGFR-3-positive. The pN0 counterparts were 4.55% and 9.09%, respectively (P < 0.05). The expressions of Twist, VEGF-C, and VEGFR-3 in pN+ patients obtained through Western blotting analysis were significantly higher than those in pN0 patients, and the levels of VEGF-C and VEGFR-3 were positively correlated with that of Twist. Twist expression correlates with lymph node metastasis. The mechanism involved in such a correlation may be related to lymphangiogenesis.Chinese medical journal 05/2011; 124(10):1483-7. · 0.86 Impact Factor -
Article: [Vocal fold augmentation by injection of autologous fascia and fat].
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ABSTRACT: To evaluate the effect of combination of autologous fascia and fat injection into vocal fold for the treatment of patients with unilateral vocal fold paralysis and to observe the long-term effectiveness of this procedure. A total of 26 unilateral vocal fold paralysis patients underwent vocal fold injection under general anesthesia, meanwhile, the mucosa of the injected point was sutured through laryngoscope under direct vision. There were 6 patients underwent autologous fat injection into vocal fold (group A), and 20 patients underwent autologous anterior rectus sheath fascia and fat injection (group B). Therapeutic efficacy were evaluated by videostroboscopy, voice-related parameters analysis and voice evaluation before and after treatment. Clinical analysis of this procedure was retrospectively performed in this serial of patients. All patients were followed up for 24 months. On the third day after operation, there was an acute inflammatory reaction induced by the graft. This reaction disappeared three months later. In all 20 cases, videolaryngostroboscopy showed significant improvement of the glottic closure, the improvement in acoustical parameters was statistically significant (P < 0.01). Perceptual evaluation of GRBAS scale showed significant improvement of phonatory function on G, B, A scale. The results remained stable 6 - 24 months after operation and were not changed by the length of follow-up. And in the 6 cases, videolaryngostroboscopy showed significant improvement of the glottic closure at 3 months compared with preoperative observation, a little spindle-shaped disclosure. The improvement in acoustical parameters was significant statistically at 3, 6 and 24 months (P < 0.05 or < 0.01), the voice quality decreased significantly at 6 and 24 months compared with 3 months (P < 0.05 or < 0.01). The significant differences were not observed between 6 and 24 months (P > 0.05). No complications were observed in all patients perioperatively or during the follow-up period. Voice-related parameters jitter, normalized noise energy and maximum phonation time showed significant differences between Group A and Group B on 24 months (P < 0.05 or < 0.01). The combination of autologous fascia and fat vocal fold injection is an effective procedure for the treatment of unilateral vocal fold paralysis, and the stable results can be achieved during the follow-up period for 24 months.Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery 04/2011; 46(4):269-74. -
Article: The probable role of tumor stem cells for lymph node metastasis in supraglottic carcinoma.
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ABSTRACT: Tumor stem cells (TSC), which are considered as likely candidates for the origin of cancer, are deduced to be responsible for tumor metastasis theoretically. We therefore investigated whether TSC were associated with lymph node metastasis in supraglottic carcinoma. Immunohistochemistry was performed for CD44, CD133, and LYVE-1 to detect TSC and lymphatic vessel density (LVD) in 66 primary supraglottic carcinoma tissue samples from 30 patients with lymph node metastasis (N+) and 36 patients without (N0). Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot were used to detect the expression of CD44 and CD133 at mRNA and protein levels in N+ and N0 primary tumors. The LVD was 22.4 ± 10.26 in 30N+ and 6.8 ± 4.09 in 36N0 samples subjected to immunohistochemistry, which was associated with their clinical nodal stages. There were 43.33% CD44-positive and 93.33% CD133-positive samples in 30N +, and 13.89% CD44-positive and 44.44% CD133-positive samples in 36N0 (P < 0.05). However, in each positive slide, there were only 5∼10% CD44-positive cells, but 70∼85% CD133-possitive cells. The expressions of CD44 and CD133 of N+ obtained through RT-PCR and Western blot were significantly higher than those of N0. These results suggest that TSC identified through CD44-positive cells in N+ were significantly higher than those in N0, indicating that TSC may be responsible for lymph node metastasis. CD133, whose expression is not restricted to TSC, may be unspecific for TSC identification in hypostatic supraglottic carcinoma.Pathology & Oncology Research 03/2011; 17(1):33-8. · 1.37 Impact Factor -
Article: Curcumin attenuates peroxynitrite-induced neurotoxicity in spiral ganglion neurons.
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ABSTRACT: The present study was designed to investigate the effect of curcumin on peroxynitrite (ONOO(-))-induced damage in rat spiral ganglion neurons (SGNs). The primary cultured rat SGNs were exposed to ONOO(-) with or without curcumin pretreatment. Cell viability was measured by MTT assay. Apoptosis was determined by Ho.33342 and propidium iodide (PI) double staining and flow cytometry. The cellular glutathione (GSH) content, superoxide dismutase (SOD) activity and malonaldehyde (MDA) levels were evaluated by spectrophotometer. The mRNA expressions of Apaf-1, Caspase-9, Caspase-3, Bcl-2, and Bax were examined by RT-PCR, while, the protein expressions of mitochondrial and cytosolic cytochrome c, Caspase-9, Caspase-3, Bcl-2 and Bax proteins were determined by Western blot respectively. The cell viability was markedly reduced, while, the apoptotic rate increased significantly after exposure of ONOO(-) (100μM) to SGNs. The activity of SOD and level of GSH were notably reduced, whereas, the MDA level was significantly increased. Pretreatment with curcumin protected SGNs against ONOO(-)-induced cell damage, declined the apoptotic rate, and improved the levels of SOD and GSH, decreased the elevation of MDA. ONOO(-) induced cytochrome c release from the mitochondria of SGNs and subsequently activated Caspase-9, Caspase-3 and cell apoptosis. Meanwhile, pretreatment with curcumin abrogated cytochrome c release, blocked activation of Caspase-3, and altered the expression of Bcl-2 family triggered by ONOO(-). Our data indicate that curcumin can attenuate ONOO(-)-induced damage in SGNs by the anti-oxidative activity as well as protect mitochondria from oxidative stress.NeuroToxicology 01/2011; 32(1):150-7. · 3.10 Impact Factor -
Article: [Nimesulide induced apoptosis and inhibited metastasis of hypopharyngeal carcinoma cells].
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ABSTRACT: To investigate the effects of Nimesulide, a selective Cox-2 inhibitor, on the apoptosis, invasion and migration of hypopharyngeal carcinoma cell line (FaDu). Viabilities of FaDu cells treated with various concentrations of Nimesulide were detected by MTT assay. Morphological changes were observed by acridine orange cytochemistry staining. Apoptosis was examined by flow cytometry. The ability of invasion and migration of cells was detected by Transwell chambers. The mRNA and protein expressions of Cox-2, MMP-9 and caspase-3 in response to Nimesulide were examined by RT-PCR and Western blot, respectively. MTT assay showed that the cell surviving rates significantly decreased in time- and concentration-dependent manners (P < 0.05). The typical morphological changes of apoptotic cells were observed. Percent of apoptosis after 6 h Nimesulide treatment was (32.4 ± 6.1)%. The metastatic cells were decreased by Nimesulide to about 20%. Nimesulide decreased the expressions of Cox-2 and MMP-9, whereas increased expression the of Caspase-3. Nimesulide could induce the apoptosis and inhibit metastasis of FaDu cells.Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery 10/2010; 45(10):854-8. -
Article: [Tubed pectoralis major myocutaneous flap for reconstruction of circumference pharyngoesophageal defects].
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ABSTRACT: To investigate the feasibility and efficacy of tubed pectoralis major myocutaneous flap in the reconstruction of circumferential defects following resection for locally advanced hypopharyngeal and cervical esophageal carcinoma. From Dec. 2004 to Oct. 2008, 30 patients underwent immediate reconstruction by tubed pectoralis major myocutaneous flap for circumferential defects following resection of primary tumours. Among them, 22 were hypopharyngeal carcinoma, 7 were cervical esophageal carcinoma and one was recurrent laryngeal carcinoma involved the hypopharyngeal lumen. Five of 30 patients had received previous radiotherapy and three had failed in the previous surgical procedure. In this series, 12 patients had total pharyngolaryngectomy and 18 had total pharyngolaryngectomy and partial cervical esophagectomy. Postoperative pharyngocutaneous fistula formation occurred in 4 patients, 2 of them with previous radiotherapy and 2 with diabetes, and the fistulae healed later. Two patients developed anastomotic strictures at the upper junction, but they had good responses to dilatation treatment and had satisfactory oral intake. The postoperative follow-up time ranged from 8 to 56 months. Median follow-up was 18 months. One-year survival rate was 71.4% and three-year survival rate was 42.5%. The tubed pectoralis major myocutaneous flap is a reliable procedure to reconstruct hypopharyngeal circumferential defects following resection of advanced hypopharyngeal and cervical esophageal carcinoma. This method may be the optimal choice for the reconstruction of hypopharyngeal circumferential defects following resection of recurrent carcinoma. The incidence of fistula and stenosis could be kept at an acceptable level.Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery 05/2010; 45(5):401-5. -
Article: Alteration in TWIST expression: possible role in paclitaxel-induced apoptosis in human laryngeal carcinoma Hep-2 cell line.
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ABSTRACT: To explore the relationship between alteration in the expression of TWIST, highly conserved transcription factor from the basic helix-loop-helix family, and apoptosis of Hep-2 cells induced by chemotherapeutic agent paclitaxel. Morphological changes of Hep-2 cells were observed by acridine orange cytochemistry staining. Viability of Hep-2 cells treated with various concentrations of paclitaxel was examined by cell proliferation assay. Apoptosis was examined by flow cytometry. The mRNA and protein expression of TWIST in response to paclitaxel at 24 hours, 48 hours, and 72 hours was examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting, respectively. Typical morphological changes of apoptotic cells at 24 hours, 48 hours, or 72 hours after treatment wiyth paclitaxel (10x10(-9) mol/L) were observed. The cell survival rates significantly decreased in a concentration- and time-dependent manner (P=0.001). Paclitaxel-induced apoptosis increased with culture time (22.6+/-5.3% after 24 hours, 38.7+/-7.9% after 48 hours, and 52.4+/-14.3% after 72 hours; P=0.002). Both mRNA and protein expression of TWIST was markedly decreased at both mRNA levels and protein levels, at 24 hours, 48 hours, and 72 hours in the paclitaxel-induced apoptosis of Hep-2 cells (P<0.001). TWIST, which has a significantly decreased expression in response to paclitaxel in Hep-2 cells, may play a pivotal role in paclitaxel-induced apoptosis of Hep-2 cells.Croatian Medical Journal 12/2009; 50(6):536-42. · 1.80 Impact Factor -
Article: [Role of TWIST in the apoptosis of Hep-2 cells induced by paclitaxel].
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ABSTRACT: To explore the relationship between the expression of transcription factor TWIST and apoptosis of Hep-2 cells induced by paclitaxel. Morphological changes of Hep-2 cells were observed by reserved microscopy and acridine orange cytochemistry staining. Viability of Hep-2 cells treated with various concentrations of paclitaxel was detected by MTT assay. Apoptosis was examined by flow cytometry. The expressions of transcription factor TWIST at both mRNA and protein level in response to paclitaxel at 24 h, 48 h and 72 h were then examined by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot, respectively. Typical morphological changes of apoptotic cells, i.e., cellular rounding-up, cytoplasmic contraction, chromatin condensation and, particularly, apoptotic body, the main morphological characteristic of apoptosis, were observed by reserved microscopy and acridine orange cytochemistry staining. The cell surviving rates significantly decreased in a concentration- and time-dependent manner as evidenced by MTT assay (P < 0.05). Percent of apoptosis after 24 h, 48 h or 72 h paclitaxel-treatment was (22.6 +/- 5.3)%, (38.7 +/- 7.9)% and (52.4 +/- 14.3)%, respectively, whereas the percent of control was (9.85 +/- 5.83)%. There existed a statistically significant difference between treatment and control (F = 12.621, P < 0.05). The expression of TWIST at both mRNA and protein levels for 24 h, 48 h or 72 h in the paclitaxel-induced apoptosis of Hep-2 cells were decreased by 16.7%, 46.8%, 76.9% (F = 10.407, P < 0.05) and 16.4%, 33.6%, 69.6% (F = 18.013, P < 0.05) respectively. TWIST, which is significantly decreased in expression in response to paclitaxel in Hep-2 cells, may play a pivotal role in paclitaxel-induced apoptosis of Hep-2 cells.Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery 09/2009; 44(9):772-6. -
Article: MDR1/P-gp and VEGF synergistically enhance the invasion of Hep-2 cells with multidrug resistance induced by taxol.
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ABSTRACT: Tumor invasion/metastasis and multidrug resistance (MDR) are the main causes of treatment failure and high mortality in all kinds of cancer patients. The relationship between the two factors is still unclear. The aim of this study is to investigate the association between MDR and invasion, especially the role of multidrug resistance 1/P-glycoprotein (MDR1/P-gp) and vascular endothelial growth factor (VEGF) during the invasion. Multidrug resistance 1 (MDR1) and VEGF receptor 2 (VEGFR-2) were detected with real-time quantitative reverse-transcription polymerase chain reaction (RT-PCR) and Western blotting at the levels of messenger RNA (mRNA) and protein, respectively. RNA interference was applied to inhibit the expression of MDR1. The invasive assays were performed with the CHEMICON cell invasion assay kit. The MDR cell line induced by Taxol (Hep-2T cell) was more invasive than its parent cell line (Hep-2 cell), which was at least in part mediated through the overexpressed MDR1/P-pg. MDR1-targeted RNA interference could effectively inhibit the expression of MDR1 and obviously decrease the invasive ability. Synergistic enhancing effects existed between MDR1/P-gp and VEGF on the invasion of Hep-2T cells. The expression of VEGFR-2 was elevated in Hep-2T cells. SU1498 could significantly decrease the invasion of Hep-2T cells. MDR1-targeted RNA interference and SU1498 had synergistic decreasing effect on the invasion of Hep-2T cells. MDR1/P-pg may be a risk predictor for the invasion of laryngeal cancer. MDR1 knock down and VEGFR-2 inhibitor may be two promising treatment regiments for advanced laryngeal carcinoma patients with MDR and invasion/metastasis.Annals of Surgical Oncology 03/2009; 16(5):1421-8. · 4.17 Impact Factor -
Article: [Recurrent laryngeal nerve decompression].
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ABSTRACT: To study the result of recurrent laryngeal nerve decompression in the treatment of functional disturbance of recurrent laryngeal nerve caused by thyroid surgery or thyroid benign tumors. From October 2002 to June 2005, 9 cases of unilateral recurrent laryngeal nerve paralysis and 4 cases of glottic insufficiency were treated with recurrent laryngeal nerve decompression. Seven cases of the nerve paralysis were caused by the surgery of benign thyroid tumors resection which were done by general surgeons. The paralysis nerve were found ligated in 6 of the 7 cases, and nerve-scar adhesion was found in the other case. Beside nerve decompression, type I thyroplasty have been undertaken in the same time to 2 of the 7 cases with severe hoarseness. One case of thyroid adenoma and 1 case of nodular goiter with unilateral recurrent laryngeal nerve paralysis were treated with tumor resection and nerve decompression respectively. Four cases of glottic insufficiency, 3 cases of nodular goiter were treated with tumor resection and nerve decompression, and Hashimoto's thyroiditis in the other case was treated with partial lobe resection and nerve decompression. The recovery of function of recurrent laryngeal nerve were detected to the recovery of vocal cord mobility through electrolaryngoscope postoperatively. For the 7 cases of recurrent laryngeal nerve paralysis after thyroid surgery, the motion of the paralysed vocal cord restored within 3 months in 6 cases with the interval of 1 week to 3 months between the two operations, no restoration was found in the other patient with an interval above 4 months between the two operations after 1 year follow-up. For the thyroid adenoma and nodular goiter with unilateral recurrent laryngeal nerve paralysis, the motion of paralysed vocal cord restored within 3 months respectively after decompression. The glottic closure recovered within 1 week after decompression in the 4 cases of the glottic insufficiency patient. Exploration and decompression as soon as possible to the paralysed recurrent laryngeal nerve because of thyroid surgery are very important for the restoration of the function of the nerve. For the patient with serve hoarseness, nerve decompression and type I thyroplasty at the same time is recommended in an effort to relieve hoarseness as soon as possible. For the functional disturbance of recurrent laryngeal nerve with thyroid neoplasm patient, early exploration and decompression of the nerve is imperative.Zhonghua er bi yan hou tou jing wai ke za zhi = Chinese journal of otorhinolaryngology head and neck surgery 07/2006; 41(6):408-11. -
Article: Overexpression of Smac/DIABLO in Hep-2 cell line: possible role in potentiating the sensitivity of chemotherapeutic drugs.
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ABSTRACT: The major obstacles for tumor chemotherapy are drug resistance and/or adverse effects on the host. In the present study we investigated the role of the second mitochondria-derived activator of caspase (Smac/DIABLO) in the action of cisplatin (DDP), 5-fluorouracil (5-FU), and the combination of both in Hep-2 cells. Hep-2 laryngeal carcinoma cells exposed to DDP, 5-FU and the combination of both were investigated. Cell viability was determined by MTT assay. Apoptosis was measured by Ho.33342 and PI double staining and flow cytometry. The expression of Smac/DIABLO at the mRNA and protein level was assayed by RT-PCR and Western blotting. DDP, 5-FU and the combination of both drugs reduced the cell survival rates in a concentration- and time-dependent manner. The drug combination not only exerted a stronger inhibitory effect, but also at a lower concentration compared with the single drugs. Apoptosis was concomitant in a caspase-dependent manner. The expression of Smac/DIABLO increased significantly at both mRNA and protein levels after cell exposure to the combination compared with single drugs. Smac/DIABLO plays a pivotal role in attaining a synergistic effect in Hep-2 cells in response to this combined strategy.Tumori 96(2):310-5. · 0.86 Impact Factor
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2011
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University of Jinan (Jinan, China)
Jinan, Shandong Sheng, China
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2006–2010
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Shandong University
Jinan, Shandong Sheng, China
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