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1.
Chin Med Sci J ; 34(3): 184-193, 2019 Sep 30.
Artigo em Inglês | MEDLINE | ID: mdl-31601301

RESUMO

Objective Our previous study has revealed that iASPP is elevated in human head and neck squamous cell carcinoma (HNSCC) and iASPP overexpression signifcantly correlates with tumor malignant progression and poor survival of HNSCC. This study investigated the function of iASPP playing in proliferation and invasion of HNSCC in vitro. Methods HNSCC cell line Tu686 transfected with Lentiviral vector-mediated iASPP-specific shRNA and control shRNA were named the shRNA-iASPP group and shRNA-NC group, respectively. The non-infected Tu686 cells were named the CON group. CCK-8 assay, flow cytometry, transwell invasion assay were performed to detect the effects of iASPP inhibition in vitro. Results Our results demonstrated that the proliferation of shRNA-iASPP cells at the time of 72 h (F=32.459, P=0.000), 96 h (F=51.407, P=0.000), 120 h (F=35.125, P=0.000) post-transfection, was significantly lower than that of shRNA-NC cells and CON cells. The apoptosis ratio of shRNA-iASPP cells was 9.42% ± 0.39% (F=299.490, P=0.000), which was significantly higher than that of CON cells (2.80% ± 0.42%) and shRNA-NC cells (3.18% ± 0.28%). The percentage of shRNA-iASPP cells in G0/G1 phase was 74.65% ± 1.09% (F=388.901, P=0.000), which was strikingly increased, compared with that of CON cells (55.19% ± 1.02%) and shRNA-NC cells (54.62% ± 0.88%). The number of invading cells was 56 ± 4 in the shRNA-iASPP group (F=84.965, P=0.000), which decreased significantly, compared with the CON group (111 ± 3) and the shRNA-NC group (105 ± 8). The survival rate of shRNA-iASPP cells administrated with paclitaxel was highly decreased, compared with CON cells and shRNA-NC cells (F=634.841, P=0.000). Conclusion These results suggest iASPP may play an important role in progression and aggressive behavior of HNSCC and may be an efficient chemotherapeutic target for the treatment of HNSCC.

2.
Technol Cancer Res Treat ; 17: 1533034617750309, 2018 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-29357787

RESUMO

Radiotherapy is the primary treatment for nasopharyngeal carcinoma while radioresistance can hinder efficient treatment. To explore the role of annexin A1 and its potential mechanisms in radioresistance of nasopharyngeal carcinoma, human nasopharyngeal carcinoma cell line CNE2-sh annexin A1 (knockdown of annexin A1) and the control cell line CNE2-pLKO.1 were constituted and CNE2-sh annexin A1 xenograft mouse model was generated. The effect of annexin A1 knockdown on the growth of xenograft tumor after irradiation and radiation-induced DNA damage and repair was analyzed. The results of immunohistochemistry assays and Western blotting showed that the level of annexin A1 was significantly downregulated in the radioresistant nasopharyngeal carcinoma tissues or cell line compared to the radiosensitive nasopharyngeal carcinoma tissues or cell line. Knockdown of annexin A1 significantly promoted CNE2-sh annexin A1 xenograft tumor growth compared to the control groups after irradiation. Moreover, the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling assays revealed that knockdown of annexin A1 significantly inhibited apoptosis in vivo compared to the control groups. We assessed the intracellular reactive oxygen species levels and the extent of radiation-induced DNA damage and repair using reactive oxygen species assay, comet assays, and immunohistochemistry assay. The results showed that knockdown of annexin A1 remarkedly reduced the intracellular reactive oxygen species levels, level of DNA double-strand breaks, and the phosphorylation level of H2AX and increased the accumulation of DNA-dependent protein kinase in nasopharyngeal carcinoma cells after irradiation. The findings suggest that knockdown of annexin A1 inhibits DNA damage via decreasing the generation of intracellular reactive oxygen species and the formation of γ-H2AX and promotes DNA repair via increasing DNA-dependent protein kinase activity and therefore improves the radioresistance in nasopharyngeal carcinoma cells. Together, our findings suggest that knockdown of annexin A1 promotes radioresistance in nasopharyngeal carcinoma and provides insights into therapeutic targets for nasopharyngeal carcinoma radiotherapy.


Assuntos
Anexina A1/genética , Apoptose/genética , Carcinoma Nasofaríngeo/genética , Tolerância a Radiação/genética , Adulto , Idoso , Animais , Anexina A1/metabolismo , Biomarcadores , Proteínas de Ligação ao Cálcio/metabolismo , Linhagem Celular Tumoral , Dano ao DNA , Modelos Animais de Doenças , Feminino , Regulação Neoplásica da Expressão Gênica , Técnicas de Silenciamento de Genes , Histonas/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Camundongos , Pessoa de Meia-Idade , Carcinoma Nasofaríngeo/metabolismo , Carcinoma Nasofaríngeo/patologia , Carcinoma Nasofaríngeo/radioterapia , Gradação de Tumores , Estadiamento de Neoplasias , Interferência de RNA , RNA Interferente Pequeno/genética , Espécies Reativas de Oxigênio/metabolismo , Ensaios Antitumorais Modelo de Xenoenxerto
3.
J Thorac Dis ; 7(9): 1650-7, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26543614

RESUMO

BACKGROUND: To investigate the prevalence of nosocomial infections, the distribution of nosocomial infection sites, the use of antibiotic and the situation of detected nosocomial infection pathogens in the Inner Mongolia Autonomous Region of China from 2012 to 2014, to grasp the current conditions of regional nosocomial infections in timely, for the development of infection prevention and control measures to provide a basis for effective hospital. METHODS: A survey of the prevalence of nosocomial infections was conducted in target hospitals using the combination of a bedside survey and medical record review. RESULTS: In total, 101,907 inpatients were surveyed from 2012 to 2014. There were 1,997 cases of nosocomial infections, accounting for an average prevalence of 1.96%. The infection site was mainly the lower respiratory tract. Higher prevalence of nosocomial infections occurred in the comprehensive intensive care unit (ICU), Neurosurgery Department, and Hematology Department. The average rate of antibiotic use was 33.72%, and the average submission rate for bacterial cultures for patients who received therapeutic treatment with antibiotics was 28.26%. The most common pathogens associated with nosocomial infections were Gram-negative (G(-)) bacteria, and frequently detected bacterial pathogens included Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Acinetobacter baumannii, and Staphylococcus aureus. CONCLUSIONS: The survey of the prevalence of nosocomial infections helped to identify problems in the control process of nosocomial infections and to develop targeted measures for the prevention and control of these infections accordingly.

4.
Medicine (Baltimore) ; 94(6): e502, 2015 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-25674742

RESUMO

Our previous study indicated overexpression of metadherin (MTDH) is an adverse prognostic factor in squamous cell carcinoma of the head and neck (SCCHN) and promotes SCCHN cell proliferation and invasion. However, its mechanism remains unclear. Recent studies have indicated that MTDH is a cancer-metastasis-associated molecule that participates in the process of angiogenesis. Therefore, the study is aimed to investigate that whether vascular endothelial growth factor (VEGF), as one of the most potent proangiogenic cytokines, is regulated by MTDH and the role of the phosphatidylinositide 3-kinases/Protein Kinase B (PI3K/Akt) pathway in this process of regulation and the clinical significance of both MTDH and VEGF in SCCHN.Immunohistochemistry was used to assay the expression of MTDH and VEGF in a cohort of 189 SCCHN patients with intact follow-up information. The expression of MTDH was then upregulated or inhibited by lentivirus-mediated MTDH Complementary deoxyribonucleic acid or MTDH short hairpin ribonucleic acid (shRNA) to observe the resulting alterations in VEGF expression and the PI3K/Akt signaling pathway in SCCHN cell lines. In addition, the PI3K/Akt pathway was modulated to observe the resulting changes in the MTDH-mediated expression of VEGF.The immunohistochemistry data showed that MTDH expression is positively correlated with VEGF expression in SCCHN tissues. Moreover, the overexpression of MTDH in SCCHN Tu686 and 5-8F cells led to increases in the expression of VEGF, and this effect was accompanied by activation of the PI3K/Akt pathway. Conversely, shRNA-mediated knockdown of MTDH led to decreased VEGF expression. In addition, inhibition of the Akt signaling pathway reversed the upregulation of VEGF resulting from MTDH overexpression. Moreover, the survival analysis revealed that VEGF is an independent prognostic factor, and a combined survival analysis based on both MTDH and VEGF showed synergistic effects in the prognosis evaluation of SCCHN patients.The findings of the present study demonstrate that MTDH regulates the expression of VEGF via the PI3K/Akt signaling pathway, indicating the potential role of the MTDH-mediated activation of VEGF signaling pathway in SCCHN angiogenesis and metastasis.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Moléculas de Adesão Celular/fisiologia , Neoplasias de Cabeça e Pescoço/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Fator A de Crescimento do Endotélio Vascular/metabolismo , Western Blotting , Carcinoma de Células Escamosas/mortalidade , Células Cultivadas , Ensaio de Imunoadsorção Enzimática , Neoplasias de Cabeça e Pescoço/mortalidade , Humanos , Imuno-Histoquímica , Prognóstico , Proteínas Proto-Oncogênicas c-akt/antagonistas & inibidores , RNA Interferente Pequeno/fisiologia , Transdução de Sinais/fisiologia , Transfecção , Regulação para Cima
5.
PLoS One ; 9(2): e88656, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-24558407

RESUMO

A number of studies have explored the association of the aldehyde dehydrogenases-2 (ALDH2) Glu487Lys polymorphism and risk of colorectal cancer; however, the results are inconsistent. We performed this meta-analysis to clarify this issue using all the available evidence. Relevant studies were retrieved by searching PubMed. Eleven case-control studies were included in the meta-analysis, representing 2909 cases and 4903 controls. The pooled results based on all included studies showed a decreased colorectal cancer risk in the analysis of the GA genotype vs. the GG genotype (OR = 0.81, 95%CI = 0.68-0.98, p = 0.03) and in the dominant genetic model analysis (OR = 0.81, 95%CI = 0.67-0.98, p = 0.03). However, there was no statistical difference in the AA vs. GG analysis (OR = 0.74, 95%CI = 0.52-1.06,p = 0.11) and the recessive genetic model analysis (OR = 0.86, 95%CI = 0.69-1.07, p = 0.17). Cumulative meta-analysis based on publication time confirmed these findings. Patients with colorectal cancer had a higher frequency of the GG genotype (OR = 1.10, 95% CI = 1.02-1.20, p = 0.02) and a lower frequency of the GA genotype (OR = 0.89, 95%CI = 0.81-0.98, p = 0.02) comparing with control population. Our results suggested that the ALDH2 Glu487Lys polymorphism may be associated with a decreased risk of colorectal cancer.


Assuntos
Aldeído Desidrogenase/genética , Neoplasias Colorretais/enzimologia , Neoplasias Colorretais/genética , Predisposição Genética para Doença/genética , Polimorfismo de Nucleotídeo Único , Aldeído-Desidrogenase Mitocondrial , Humanos
6.
Artigo em Chinês | MEDLINE | ID: mdl-24195824

RESUMO

OBJECTIVE: To investigate the irradiation induced epithelial-mesenchymal transition (EMT) in nasopharyngeal carcinoma (NPC) in vitro. METHODS: NPC CNE-2 cells with radioresistance (CNE-2-Rs) were established by exposure to gradiently increased dose of irradiation. CCK-8 cell viability kits, colony formation assay and fluorescence-activated cell sorting analysis were used to confirm the capacity of radioresistance of CNE-2-Rs cells. Invert microscope was used to monitor the morphological changes and western blot was applied to detect the expression of epithelial cell marker E-cadherin and mesenchymal cell marker Vimentin during the phase of CNE-2 exposure to irradiation. RESULTS: Irradiation exposure successfully induced the radioresistance of CNE-2 cells. After exposed to irradiation, the survival rate in CNE-2-Rs was higher than that in CNE-2 by CCK-8 assays. No significant difference of proliferation ability was observed between the CNE-2 and CNE-2-Rs pre-radiotherapy, but a higher proliferation ability in the CNE-2-Rs post-radiotherapy. By using the colony forming assay, the parameters of CNE-2 and CNE-2-Rs in multi-target single-hit and linear quadratic model were obtained and the data demonstrated that parameters mean lethal dose (D0) , quasi-thres hold dose (Dq) , surrival fraction in 2Qy (SF2) and mean inctivation dose (MID) value increased, α and α/ß value decreased (P < 0.05) . At the same time, the CNE-2-Rs cells showed higher percentage of cells in S and G2 phase (P < 0.05) . In terms of biomorphology, CNE-2-Rs cells were more narrow, long strips or fusiform shapes, stretched out tentacles, and the contacts between them were loosened. When radiation dose accumulated to 24 Gy, an over-expression of Vimentin was observed in treated cells, while E-cadherin was down-regulated (P < 0.01) . CONCLUSIONS: NPC cells present with typical morphorlogical and biomolecular changes of EMT during exposure to irradiation, indicating the potential critical roles of EMT in the malignant behavior of radioresistance in NPC.


Assuntos
Transição Epitelial-Mesenquimal , Neoplasias Nasofaríngeas/radioterapia , Caderinas , Carcinoma , Linhagem Celular Tumoral , Sobrevivência Celular , Regulação para Baixo , Citometria de Fluxo , Humanos , Técnicas In Vitro , Carcinoma Nasofaríngeo , Vimentina/metabolismo
7.
Artigo em Chinês | MEDLINE | ID: mdl-23755789

RESUMO

OBJECTIVE: To investigate the regulatory effect of erythropoietin-producing hepatocellular receptor (EphA2) on the expression of VEGF protein, a pro-angiogenic factor, via p38 mitogen-activated protein kinase (p38 MAPK) signaling pathway in squamous cell carcinoma of the head and neck(SCCHN) in vitro. METHODS: SCCHN Tu686 cells were transfected with EphA2 overexpression vector pEGFP-N1-EphA2. Western blot was used to detect the expression of p38 MAPK and enzyme-linked immunosorbent assay (ELISA) was applied to assay of VEGF. SB203580 as a inhibitor of p38 MAPK signaling pathway was used. RESULTS: The expression of VEGF protein was significantly up-regulated in Tu686 cells transfected with EphA2 overexpression vector (535.31 ± 45.71) pg/ml, when compared with Tu686 cells transfected with empty vector (400.99 ± 33.50) pg/ml and Tu686 cells with no transfection (385.30 ± 33.50) pg/ml (F = 17.091, P < 0.01). The expression of phosphorylated p38 MAPK was obviously increased in Tu686 cells with EphA2 overexpression. SB203580 inhibited the expressions of VEGF and phosphorylated p38 MAPK proteins in Tu686 cells with EphA2 overexpression. CONCLUSION: EphA2 can regulate the expression of VEGF protein and stimulate p38 MAPK signaling pathway.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Neoplasias de Cabeça e Pescoço/metabolismo , Sistema de Sinalização das MAP Quinases , Receptor EphA2/fisiologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Linhagem Celular Tumoral , Inibidores Enzimáticos/farmacologia , Humanos , Imidazóis/farmacologia , Piridinas/farmacologia , Carcinoma de Células Escamosas de Cabeça e Pescoço , Proteínas Quinases p38 Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
8.
Zhonghua Bing Li Xue Za Zhi ; 42(2): 111-5, 2013 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-23710918

RESUMO

OBJECTIVE: To assess the protein expression of astrocyte elevated gene-1 (AEG-1) in tissue specimens of laryngeal squamous cell carcinoma (LSCC), and to correlate its expression with clinicopathological parameters and prognosis in patients with LSCC. METHODS: RT-PCR was used to assay the expression of AEG-1 mRNA in 13 pairs of LSCC tissues and their corresponding noncarcinoma epithelia. Immunohistochemistry was performed on paraffin-embedded tissue specimens to investigate the protein expression of AEG-1 in 88 cases of LSCC specimens and 15 cases of adjacent epithelial samples. RESULTS: The expression of AEG-1 mRNA was significantly increased in LSCC tissues compared to adjacent noncarcinoma epithelial tissues (0.81 ± 0.17 vs. 0.23 ± 0.10;t = 10.337, P < 0.001). Meantime, the positive rate of AEG-1 protein in 88 cases of LSCC was 87.5% (77/88). However, 15 cases of adjacent noncarcinoma epithelial merely demonstrated negative or mild expression of AEG-1 protein. AEG-1 overexpression was closely correlated with T stage (χ(2) = 6.289, P = 0.018), clinical stage (χ(2) = 11.049, P < 0.01), metastasis (χ(2) = 20.859, P < 0.01) and recurrence(χ(2) = 13.459, P < 0.01). The overall survival rates of patients with AEG-1 overexpression and low expression were 35.9% and 86.4%, respectively (χ(2) = 23.409, P < 0.01). Multivariate Cox regression analysis revealed that AEG-1 expression was an independent prognostic factor (P = 0.016). CONCLUSION: AEG-1 protein may play a critical role in the initiation and progression of LSCC, implicating its predictive value in prognosis.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Moléculas de Adesão Celular/metabolismo , Neoplasias Laríngeas/metabolismo , Neoplasias Laríngeas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/genética , Carcinoma de Células Escamosas/cirurgia , Moléculas de Adesão Celular/genética , Feminino , Seguimentos , Humanos , Neoplasias Laríngeas/genética , Neoplasias Laríngeas/cirurgia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Modelos de Riscos Proporcionais , RNA Mensageiro/metabolismo , Taxa de Sobrevida
9.
Artigo em Chinês | MEDLINE | ID: mdl-23141445

RESUMO

OBJECTIVE: To study the effect and molecular mechanism of epigallocatechin-3-gallate (EGCG) on epithelial-mesenchymal transition (EMT) in vitro induced by human recombinant TGF-ß1 protein in squamous cell carcinoma of the head and neck. METHODS: EMT morphological changes of Tu686 cells were observed after sequential treatment of 5 ng/ml TGF-ß1 and 20 µmol/L EGCG. Tu686 cells were collected after the treatment of 5 ng/ml TGF-ß1 for 24 h and EGCG with different concentrations (0, 10, 20, 30 µmol/L) for another 24 h or 20 µmol/L EGCG treatment for different time phase (6, 12, 24 h). Then RT-PCR and Western-blot were applied to detect mRNA and protein expression level of epithelial cell marker E-cadherin, mesenchymal cell marker Vimentin and Smad7, an inhibit molecule of TGF-ß1 mediated pathway in Tu686 cells. RESULTS: TGF-ß1 successfully induced characterized EMT morphological and molecular changes in Tu686 cells, in which expression of E-cadherin decreased, Vimentin increased and Smad7 declined. However, EGCG could reverse the TGF-ß1 mediated process of EMT by downregulating the expression of Vimentin and upregulating the expression of E-cadherin and Smad7. CONCLUSION: EGCG significantly inhibits TGF-ß1-mediated EMT inTu686 cell lines of SCCHN, which maybe associated with the upregulated-expression of Smad7, an inhibitor in TGF-ß1 signaling pathway.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Catequina/análogos & derivados , Transição Epitelial-Mesenquimal/efeitos dos fármacos , Neoplasias de Cabeça e Pescoço/metabolismo , Antígenos CD , Caderinas/metabolismo , Catequina/farmacologia , Linhagem Celular Tumoral , Humanos , Transdução de Sinais/efeitos dos fármacos , Proteína Smad7/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço , Fator de Crescimento Transformador beta1/metabolismo , Vimentina/metabolismo
10.
Artigo em Chinês | MEDLINE | ID: mdl-22932142

RESUMO

OBJECTIVE: To investigate the mRNA and protein expressions of high mobility group box-1 (HMGB1) in the tumor tissues and sera of patients with laryngeal squamous cell carcinoma (LSCC) and their clinical significance. METHODS: Semi-quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were used to detected the expressions of HMGB1 mRNA and protein in the tumors and adjacent normal epithelial tissues in 30 patients with LSCC. Serum HMGB1 protein levels in the patients with LSCC and in 10 healthy volunteers were detected with enzyme-linked immunosorbent adsorption experiment (ELISA). RESULTS: RT-PCR demonstrated that the mean relative mRNA expression levels of HMGB1 (HMGB1/GAPDH) in LSCC tissues and in adjacent normal epithelial tissues were 1.25 ± 0.12 and 0.32 ± 0.04, respectively (t = 40.27, P < 0.05). Western blot revealed that the mean relative protein expression levels of HMGB1 (HMGB1/ß-actin) were 1.29 ± 0.10 and 0.34 ± 0.03 (t = 49.84, P < 0.05), respectively. Both mRNA and protein expression levels of HMGB1 were associated with T stage, clinical stage, lymph node metastasis status and smoking (all P < 0.05), but no significant correlation with age, alcohol consumption and primary tumor grade and location (all P > 0.05). Mean serum HMGB1 protein levels in patients with LSCC and healthy volunteers were (24.80 ± 14.08) ng/ml and (23.58 ± 14.69) ng/ml (t = 0.37, P > 0.05). CONCLUSIONS: Both mRNA and protein expressions of HMGB1 were obviously elevated in LSCC, which were associated closely with T stage, clinical stage and lymph node metastasis.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Proteína HMGB1/metabolismo , Neoplasias Laríngeas/metabolismo , Adulto , Idoso , Carcinoma de Células Escamosas/sangue , Carcinoma de Células Escamosas/patologia , Estudos de Casos e Controles , Feminino , Proteína HMGB1/sangue , Proteína HMGB1/genética , Humanos , Neoplasias Laríngeas/sangue , Neoplasias Laríngeas/patologia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , RNA Mensageiro/sangue , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
11.
Zhonghua Zhong Liu Za Zhi ; 34(2): 132-6, 2012 Feb.
Artigo em Chinês | MEDLINE | ID: mdl-22780932

RESUMO

OBJECTIVE: To evaluate the expression of HMGB1 protein in tissue specimens of laryngeal squamous cell carcinoma (LSCC) and adjacent normal mucosa, and explore the correlation of HMGB1 protein expression with clinicopathologic features and prognosis in LSCC. METHODS: Ninty-three cases of LSCC and 5 cases of adjcent mucosal tissue samples were included in this study. Immunohistochemical staining was performed on paraffin-embedded tissue specimens to examine the HMGB1 protein expression. The data were futher correlated with the clinicopathological features and prognosis of the LSCC patients. RESULTS: The positive rates of HMGB1 expression in LSCC specimens was 87.1%, significantly higher than that in the adjcent normal mucosa samples (46.7%, P = 0.001), and its overexpresion was closely correlated with T stage (Chi2 = 10.878, P = 0.004), clinical stage (Chi2 = 21.115, P < 0.01), metastasis (Chi2 = 28.298, P < 0.01) and recurrence (Chi2 = 14. 923, P = 0.001) in patients with LSCC. Patients with HMGB1 overexpression had both poorer disease-free survival and poorer overall survival compared with that in patients with low HMGB1 expression (Chi2 = 13.815, Chi2 = 11.912; Both P < 0.01). Univariate and multivariate Cox regression analyses revealed that HMGBI expression is an independent prognostic factor for patients with LSCC. CONCLUSIONS: The results of this study demonstrate that HMGB1 protein expression is significantly increased in LSCC tissues, and HMGB1 protein overexpression is associated with a poorer prognosis in patients with LSCC. These results suggest that HMGB1 may play a critical role in the initiation and progression of LSCC, implicating HMGB1 may become a valuable marker for the prediction of prognosis in patients with LSCC.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Proteína HMGB1/metabolismo , Neoplasias Laríngeas/metabolismo , Neoplasias Laríngeas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/cirurgia , Intervalo Livre de Doença , Feminino , Seguimentos , Humanos , Neoplasias Laríngeas/cirurgia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Modelos de Riscos Proporcionais , Taxa de Sobrevida
12.
Artigo em Chinês | MEDLINE | ID: mdl-22455776

RESUMO

OBJECTIVE: To investigate the effects of EphA2 on the angiogenesis and cervical lymph node metastasis of squamous cell carcinoma of the head and neck (SCCHN) in vivo. METHODS: EphA2 short hairpin (shRNA) lentiviral particles were used to knockdown the expression of EphA2 in SCCHN cell line M2 with high lymph nodes metastasis rate. Stable clones, obtained by puromycin screening, were assayed by RT-PCR and Western blot to validate the gene silencing efficiency and were used to establish SCCHN metastatic xenograft mouse model. Hematoxylin-eosin staining was applied to identify cervical lymph node metastasis of SCCHN in xenografted tumors. Immunohistochemistry was used to observe microvessel density. Western blot was used to investigate the protein expressions of EphA2 and vascular endothelial, growth factor (VEGF). RESULTS: EphA2 shRNA lentiviral particles efficiently decreased the mRNA and protein expressions of EphA2 in SCCHN cell line M2, which were further successfully utilized to establish SCCHN metastatic xenograft mouse model. Compared with xenografted tumors in control group, xenografted tumors in M2EphA2RNAi(+) group decreased significantly tumor volume [(430.7 ± 190.0) mm(3) (x(-) ± s) vs (1179.0 ± 289.4) mm(3)] and weight [(0.26 ± 0.10) g vs (0.54 ± 0.12) g] (both P < 0.05). More importantly, bilateral cervical lymph node metastasis rate in M2EphA2RNAi(+) was also greatly declined (Mann-Whitney U = 10.0, P < 0.05). Decreased protein expressions of EphA2 and VEGF and microvessel density were observed in M2EphA2RNAi(+) group (t = 26.751, P < 0.01). CONCLUSIONS: Knockdown of EphA2 expression led to the inhibition of tumor growth and metastasis in SCCHN nude mouse model. More importantly, SCCHN angiogenesis was also impeded, which might be associated with the decreased expression of VEGF.


Assuntos
Carcinoma de Células Escamosas/patologia , Neoplasias de Cabeça e Pescoço/patologia , Neovascularização Patológica , Receptor EphA2/genética , Animais , Linhagem Celular Tumoral , Inativação Gênica , Humanos , Metástase Linfática , Camundongos , Camundongos Nus , Prognóstico , RNA Interferente Pequeno , Carcinoma de Células Escamosas de Cabeça e Pescoço
13.
J Proteome Res ; 11(2): 1100-7, 2012 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-22070577

RESUMO

The present study was to identify and quantitate differentially expressed proteins in laryngeal squamous cell carcinoma (LSCC) tissues with or without lymph node metastasis and to explore transcriptional factors and regulation networks associated with the process. Tissue specimens were taken from 20 patients with LSCC, including 10 cases of LSCC without metastasis LSCC (N0) and 10 cases of LSCC with metastasis LSCC (Nx). Among the 643 unique proteins identified by using iTRAQ labeling and quantitative proteomic technology, 389 proteins showed an abundance change in LSCC (Nx) as compared to LSCC (N0). Cytoskeleton remodeling, cell adhesion, and immune response activation were found to be the main processes in LSCC metastasis. The construction of transcription regulation networks identified key transcription regulators for lymph node metastasis of LSCC, including Sp1, c-myc, and p53, which may affect LSCC metastasis through the epithelial-mesenchymal transition. Furthermore, our results suggest that ubiquitination may be a critical factor in the networks. The present study provides insights into transcriptional factors and regulation networks involved in LSCC metastasis, which may lead to new strategies for treatment of LSCC metastasis.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Neoplasias de Cabeça e Pescoço/metabolismo , Neoplasias Laríngeas/metabolismo , Fatores de Transcrição/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/patologia , Adesão Celular , Citoesqueleto/metabolismo , Regulação para Baixo , Feminino , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Neoplasias Laríngeas/patologia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Mapas de Interação de Proteínas , Proteínas Proto-Oncogênicas c-myc/metabolismo , Fator de Transcrição Sp1/metabolismo , Carcinoma de Células Escamosas de Cabeça e Pescoço , Proteína Supressora de Tumor p53/metabolismo , Regulação para Cima
14.
Arch Med Res ; 42(2): 128-37, 2011 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-21565626

RESUMO

BACKGROUND AND AIMS: Bone morphogenetic proteins (BMPs) have recently been shown to be involved in the genesis and progression of a wide variety of carcinomas. The present study was undertaken to estimate the effect of BMP-4 on squamous cell carcinoma of the head and neck (SCCHN) in tissue and cell levels. METHODS: In this study, immunohistochemistry, Western blotting and RT-PCR were utilized to detect the expression of BMP-4, Smad1 and phosphorylated Smad1 in SCCHN tissues or SCCHN cell lines. Those three proteins in tissues were further correlated with prognosis of SCCHN by Kaplan-Meier analysis. The epithelial-mesenchymal transition (EMT)-associated changes in SCCHN cells were detected after stimulation by human BMP-4 recombinant protein and knockdown of Smad1 gene. Meanwhile, the effect on invasiveness and migration was evaluated by invasion and scratch assays, respectively. RESULTS: BMP-4 and p-Smad1 protein were overexpressed in SCCHN tissues with cervical lymph node metastasis, which was significantly higher than those without metastasis. The expression of BMP-4 and p-Smad1 protein was negatively correlated with the prognosis of SCCHN. BMP-4 promoted the invasiveness and migration through EMT, which was demonstrated by morphological alterations, loss of E-cadherin, increase of vimentin and activation of the Smad1 signal pathway. Knockdown of Smad1 expression suppressed BMP-4 induced EMT in both cell lines and weakened the invasiveness and migration of Tu686 and Tu212 in vitro. CONCLUSIONS: Our results demonstrate that BMP-4 protein may contribute to the malignant metastasis of SCCHN, which presents as a novel prognostic marker and a potential therapeutic target for patients with SCCHN.


Assuntos
Proteína Morfogenética Óssea 4/metabolismo , Carcinoma/metabolismo , Transição Epitelial-Mesenquimal , Neoplasias de Cabeça e Pescoço/metabolismo , Invasividade Neoplásica , Neoplasias de Células Escamosas/metabolismo , Transdução de Sinais , Proteína Smad1/metabolismo , Proteína Morfogenética Óssea 4/farmacologia , Caderinas/genética , Caderinas/metabolismo , Carcinoma/mortalidade , Carcinoma/patologia , Carcinoma de Células Escamosas , Linhagem Celular Tumoral , Movimento Celular/efeitos dos fármacos , Movimento Celular/genética , Forma Celular/efeitos dos fármacos , Forma Celular/genética , Intervalo Livre de Doença , Transição Epitelial-Mesenquimal/genética , Seguimentos , Expressão Gênica/efeitos dos fármacos , Neoplasias de Cabeça e Pescoço/mortalidade , Neoplasias de Cabeça e Pescoço/patologia , Humanos , Estimativa de Kaplan-Meier , Neoplasias de Células Escamosas/mortalidade , Neoplasias de Células Escamosas/patologia , Prognóstico , Interferência de RNA , Proteína Smad1/genética , Carcinoma de Células Escamosas de Cabeça e Pescoço , Vimentina/genética , Vimentina/metabolismo
15.
Zhonghua Zhong Liu Za Zhi ; 33(3): 197-201, 2011 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-21575519

RESUMO

OBJECTIVE: To evaluate the expression of EphA2 protein in tissue specimens and cell lines of laryngeal squamous cell carcinoma (LSCC), and to further study the correlation of EphA2 protein expression with clinicopathological characteristics and prognosis in LSCC. METHODS: Western blot was applied to assess the EphA2 protein expression in LSCC cell line Hep-2 cells and the head and neck immortalized epithelial cell line NP-69 cells. Immunohistochemical staining was performed on paraffin sections of 88 cases of LSCC specimens and 16 cases of adjcent normal tissue samples to investigate the EphA2 protein expression, and to futher elucidate its correlation with clinicopathological characteristics. RESULTS: Compared with the NP-69 cells, EphA2 expression in LSCC cell line Hep-2 cells was upregulated. The positive rates of EphA2 expression in LSCC and adjcent normal tissues samples were 80.7% and 43.8%, respectively, with a significant difference between the two groups (P < 0.001). EphA2 overexpresion was closely correlated with clinical stage (I + II/III + IV, P = 0.005), metastasis (P = 0.025) and recurrence (P = 0.021) in LSCC. Furthermore, patients with EphA2 overexpression had poorer tumor-free survival and 5-year overall survival compared with that in patients with low EphA2 expression (33.3% vs. 63.2%, P = 0.003; 46.7% vs. 81.6%, P = 0.002). EphA2 expression combined with clinical stage provided a better predictive value in prognosis. Univariate and multivariate Cox regression analysis revealed that EphA2 expression is an independent prognostic factor for patients with LSCC (P = 0.019). CONCLUSIONS: The results of this study demonstrate that EphA2 protein expression is significantly increased in LSCC tissues and cell lines, and EphA2 protein overexpression is associated with tumor recurrence, metastasis and poorer prognosis in LSCC patients. These results suggest that EphA2 may play a critical role in the initiation and progression of LSCC, implicating EphA2 as a valuable marker for the prediction of recurrence, metastasis and prognosis in LSCC.


Assuntos
Carcinoma de Células Escamosas/metabolismo , Carcinoma de Células Escamosas/patologia , Neoplasias Laríngeas/metabolismo , Neoplasias Laríngeas/patologia , Receptor EphA2/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/cirurgia , Linhagem Celular , Linhagem Celular Tumoral , Intervalo Livre de Doença , Células Epiteliais/metabolismo , Feminino , Seguimentos , Humanos , Neoplasias Laríngeas/cirurgia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Pescoço , Recidiva Local de Neoplasia , Estadiamento de Neoplasias , Modelos de Riscos Proporcionais , Taxa de Sobrevida
16.
Artigo em Chinês | MEDLINE | ID: mdl-21092676

RESUMO

OBJECTIVE: To study the effects of rapamycin on cell growth and cell cycle in CNE-1 and CNE-2 cells. METHODS: Growth inhibition effect of rapamycin on CNE-1 and CNE-2 cells were assessed by cell counting kit-8 (CCK-8) assay. Morphological alterations of the cells were observed by microscope. Cell cycle and cell apoptosis were analyzed by FCM. The expression of mammalian target of rapamycin (mTOR) was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: The growth of CNE-1 and CNE-2 cells was inhibited significantly by rapamycin dose-dependently. FCM showed that CNE-1 and CNE-2 cells at 48 hours after rapamycin (150 nmol/L) treatment were arrested in the G0/G1 phase of cell cycle. However rapamycin treatment did not significantly induce apoptosis of CNE-1 and CNE-2 cells (P > 0.05). RT-PCR showed that rapamycin significantly inhibited mRNA expression of mTOR in CNE-2 cells (t = 10.625, P < 0.01). CONCLUSIONS: Rapamycin inhibits the growth of CNE-1 and CNE-2 cells by inhibiting the progression of cell cycle, which could be achieved through decreasing the expression of mTOR.


Assuntos
Ciclo Celular/efeitos dos fármacos , Sirolimo/farmacologia , Serina-Treonina Quinases TOR/antagonistas & inibidores , Linhagem Celular Tumoral , Humanos , Transdução de Sinais/efeitos dos fármacos
17.
Chin Med J (Engl) ; 122(4): 437-43, 2009 Feb 20.
Artigo em Inglês | MEDLINE | ID: mdl-19302751

RESUMO

BACKGROUND: Microarray and microdissection techniques were being used for many applications to study the carcinogenesis of some human tumors. But seldom studies had hitherto combined these two techniques to study carcinogenesis mechanism of nasopharyngeal carcinoma (NPC). To identify a set of genes involved in the carcinogenesis and development of NPC, we used the microdissected homogeneous NPC tissue cells and the pure normal epithelium pillar cells to construct the whole human genome expression profiles. METHODS: We preserved the tissue samples from nasopharynx of 18 patients (including 13 samples of NPC and 5 samples of normal or inflammatory mucous tissue samples from nasopharynx) in RNAlater Stabilization Reagent. The tissue samples were microdissected to harvest the homogeneous tissue cells, then total RNA was isolated from them. The sufficient antisense RNA (aRNA) was amplified from these total RNA. HG-U133.Plus.2.0 GeneChip was used to construct the human whole genome expression profiling of each sample. Differential patterns of expression of genes correlated with the carcinogenesis, classification and progression of NPC were identified with comparing the expression profiling data respectively in leave one out cross-validation analysis. Correlation between aRNA expression measured by the microarrays and semi-quantitative reverse transcription polymerase chain reaction (sqRT-PCR) were also ascertained, and found that hybridization results were validated in all of the 18 patients. RESULTS: Differential patterns of expression of 127 genes correlated with the carcinogenesis (A P value less than 0.001 with the 2-fold differentiated expression between case group and control group) of the NPC were filtered. The top most up-regulated and down-regulated 8 genes by the way of permutation test were also selected and listed in the paper. Expression of genes E2F6 and TSPAN-1 was identified using aRNA by sqRT-PCR and showed that there was significant difference between the average value of case groups and that of control group respectively (t = 2.170, df = 16, P = 0.045 and t = -2.946, df = 16, P = 0.009). CONCLUSIONS: We had identified some genes which could be the molecular marker during the carcinogenesis and the development of the NPC. The genes which selected from the different subgroups seemed to be implicated for the diagnosis,classification, and progression of NPC, and provided important insights into their underlying biology.


Assuntos
Microdissecção/métodos , Neoplasias Nasofaríngeas/genética , Nasofaringe/metabolismo , Análise de Sequência com Séries de Oligonucleotídeos/métodos , Adolescente , Adulto , Idoso , Epitélio/metabolismo , Feminino , Perfilação da Expressão Gênica/métodos , Regulação Neoplásica da Expressão Gênica/genética , Regulação Neoplásica da Expressão Gênica/fisiologia , Humanos , Masculino , Pessoa de Meia-Idade , Nasofaringe/citologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Adulto Jovem
18.
Arch Med Res ; 40(2): 114-23, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19237021

RESUMO

BACKGROUND AND AIMS: Genetic alteration associated with initiation and progression of laryngeal squamous cell carcinoma (LSCC) is largely unknown. The aim of this study was to identify genetic changes associated with the disease pathogenesis and pinpoint genes whose expression is impacted by these genetic alterations. METHODS: Tumor cells were collected from eight matched pairs of specimens of glottic carcinoma of the larynx and histologically normal epithelium tissues adjacent to the carcinoma by laser capture microdissection (LCM). RNAs prepared from these cells were used for genome-wide transcriptome analysis by probing 16 cDNA microarrays. Real-time quantitative RT-PCR and immunohistochemistry of tissue microarrays were used to validate a group of the differentially expressed genes identified by the cDNA microarrays. RESULTS: Hierarchical cluster analysis of the expressed genes showed that 2351 genes were differentially expressed and could distinguish cancerous and noncancerous samples. We also found 761 differentially expressed genes that were consistently different between early stage and later stage specimens. Furthermore, abnormal expression of some relevant genes such as MMP12, HMGA2, and TIMP4 were validated by real-time quantitative RT-PCR and immunohistochemistry. Analysis of gene ontology and pathway distributions then highlighted genes that may be critically important to laryngeal carcinogenesis. CONCLUSIONS: Our results suggest that using LCM plus DNA microarray analysis may facilitate the identification of clinical molecular markers for disease and novel potential therapeutic targets for LSCC.


Assuntos
Biomarcadores Tumorais/genética , Carcinoma de Células Escamosas/genética , Perfilação da Expressão Gênica , Neoplasias Laríngeas/genética , Análise de Sequência com Séries de Oligonucleotídeos , Idoso , Regulação para Baixo/genética , Feminino , Proteína HMGA2/metabolismo , Humanos , Imuno-Histoquímica , Masculino , Metaloproteinase 12 da Matriz/metabolismo , Microdissecção , Pessoa de Meia-Idade , Inibidores Teciduais de Metaloproteinases/metabolismo , Regulação para Cima/genética
19.
Artigo em Chinês | MEDLINE | ID: mdl-19035265

RESUMO

OBJECTIVE: To examine the gene expression profile of laryngeal squamous cell carcinoma (LSCC) by combination of laser capture microdissection (LCM) and microarray and to identify genetic changes in disease pathogenesis. METHODS: The study analysed 8 matched pairs of specimens of glottic carcinoma of larynx and histologically normal epithelium tissues adjacent to the carcinoma preserved in the RNA later reagent. A genome-wide transcriptome analysis was performed by probing 16 cDNA microarrays with fluorescent-labeled amplified RNA derived from laser capture microdissected cells. Real-time quantitative (RT-PCR) of tissue microarray was used to validate the reliability of cDNA microarrays. RESULTS: Significant analysis of microarray (SAM) software and hierarchical cluster analysis of the expressed genes showed that 2351 genes was significantly expressed respectively according to different analysis method (false discover rate = 0.63%). A selected set of MMP12, KRT16, RARB, PRB1 genes was identified to be consistent with array data by RT-PCR. CONCLUSIONS: The analysis of gene ontology and pathway distributions futher highlighted genes that may be critically important to laryngeal carcinogenesis. The results strongly suggest that this new approach may facilitate the identification of clinical molecular markers of disease and novel potential therapeutic targets for LSCC.


Assuntos
Carcinoma de Células Escamosas/genética , Perfilação da Expressão Gênica/métodos , Neoplasias Laríngeas/genética , Idoso , Regulação Neoplásica da Expressão Gênica , Genoma , Glote , Humanos , Lasers , Masculino , Microdissecção/métodos , Pessoa de Meia-Idade , Análise de Sequência com Séries de Oligonucleotídeos/métodos
20.
Zhonghua Yi Xue Za Zhi ; 87(36): 2541-3, 2007 Sep 25.
Artigo em Chinês | MEDLINE | ID: mdl-18067828

RESUMO

OBJECTIVE: To investigate the expression of desmoglein 3 (DSG3), a candidate target gene in the antisense RNA (aRNA) from the purified nasopharyngeal tissues in nasopharyngeal carcinoma (NPC). METHODS: Specimens of nasopharyngeal tissues were harvested from 22 NPC patients, aged 44 +/- 11 (NPC group), and 12 normal persons or patients with nasopharyngeal infectious diseases, aged 46 +/- 14. Microdissection technique was used to get homogenous tissue cells from which total RNA was isolated (control group). aRNA was amplified from the total RNA by "in vitro transcription" (IVT). The expression of DSG3 gene was identified using these aRNA by semi-quantitative reverse transcriptase polymerase chain reaction (sqRT-PCR). RESULTS: The average expression level of DSG3 in the NPC group was 3.536 +/- 2.689, significantly higher than that of the control group (0.95 +/- 0.23, df = 32, t = 3.307, P = 0.002). The expression level of DSG3 in the whole expression profiling of the NPC group was 1.06 +/- 1.60, significantly higher than that of the control group (0.48 +/- 0.23, df = 16, t = 2.145, P = 0.048). CONCLUSION: The whole genome expression profiling detected by sqRT-PCR can be used to shift the marker genes from biopsy tissue samples. DSG3 may be a tumor candidate gene in NPC.


Assuntos
Desmogleína 3/genética , Regulação Neoplásica da Expressão Gênica , Neoplasias Nasofaríngeas/genética , Adolescente , Adulto , Idoso , Feminino , Predisposição Genética para Doença , Humanos , Masculino , Pessoa de Meia-Idade , Neoplasias Nasofaríngeas/patologia , RNA Antissenso/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos
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