RESUMO
The goal of this study was to explore the molecular mechanism of hypoxia inducible factor-1 alpha (HIF-1 alpha) action on migration and invasion of esophageal carcinoma cells. We used cobalt chloride (CoCl(2) ) to mimic tumor hypoxic microenvironment and analyzed the expressions of E-cadherin, matrix metalloproteinase-2 (MMP-2), and HIF-1 alpha in esophageal carcinoma cells under hypoxia by reverse transcription polymerase chain reaction and Western blotting. To analyze the function of HIF-1 alpha in Eca109 and TE1 cells, we established stable HIF-1 alpha knockdown cells using small interfering RNA. Blocking effect was detected by Western blotting. The concentrations of MMP-2 protein in the conditioned medium were also determined by enzyme-linked immunosorbent assay. Wound-healing and cell invasion assay were used to evaluate the migration and invasion of esophageal carcinoma cells. After exposure to hypoxia, expressions of HIF-1 alpha protein in Eca109 and TE1 cells were upregulated, both mRNA and protein levels of E-cadherin were downregulated, and MMP-2 were upregulated (P < 0.05), whereas HIF-1 alpha mRNA had no significant change (P > 0.05). Small interfering RNA could block HIF-1 alpha effectively under hypoxia, then enhanced E-cadherin expression and inhibited MMP-2 expression, respectively. Furthermore, expression of HIF-1 alpha protein was stable even though MMP-2 repressed by BB2516. Compared with that in normoxia, Snail expression was enhanced when Eca109 or TE1 cells exposed to hypoxia. Once HIF-1 alpha blocked, Snail expressions were inhibited accordingly. Wound recovery and the number of invading cells decreased (P < 0.05) after HIF-1 alpha blocked. The hypoxia suppresses E-cadherin expression and enhances MMP-2 expression favoring esophageal carcinoma migration and invasion via HIF-1 alpha activation. Our observations suggest that HIF-1 alpha inhibition might be an effective strategy to weaken the migration and invasion of esophageal carcinoma cells.
Assuntos
Caderinas/metabolismo , Movimento Celular/fisiologia , Neoplasias Esofágicas/genética , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Western Blotting , Caderinas/genética , Linhagem Celular Tumoral/metabolismo , Linhagem Celular Tumoral/patologia , Movimento Celular/genética , Regulação para Baixo , Ensaio de Imunoadsorção Enzimática , Neoplasias Esofágicas/patologia , Regulação Neoplásica da Expressão Gênica , Humanos , Subunidade alfa do Fator 1 Induzível por Hipóxia/metabolismo , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 2 da Matriz/metabolismo , Invasividade Neoplásica/patologia , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sensibilidade e Especificidade , Regulação para CimaRESUMO
OBJECTIVE: We conducted this study to analyze the mechanism behind the high coagulation state induced by circulating plasma microparticle tissue factor (MP-TF) in patients with breast cancer. PATIENTS AND METHODS: 87 cases of breast cancer patients (10 cases of TNM stage I, 16 cases of II, 32 cases of III, 29 cases of IV; 8 cases of pathological type in situ carcinoma, 15 cases of ductal carcinoma, 64 cases of invasive cancer) were used as the observation group and 20 cases of benign breast lesions were used as the control group to compare MP-TF levels of plasma and coagulation parameters including prothrombin time (PT), activated partial thromboplastin time (APTT), fibrinogen (FIB) and D-dimer body (D-D) level and NF-κB signaling pathway index including P50, p65, TAK1 and IκBα levels. RESULTS: The plasma MP-TF level in the observation group was significantly higher than that in control group, and the level of MP-TF in the observation group increased with an increase in depth of TNM stage and tumor invasion; differences were statistically significant (p<0.05). In the observation group, the plasma PT and APTT were shortened, and the levels of FIB and D-D were increased; the differences were statistically significant (p<0.05). In the observation group, the levels of P50, p65, TAK1, IκBαin circulating blood were higher than those in control group; the differences were statistically significant (p<0.05). After the Pearson test, the plasma levels of MP-TF in patients with breast cancer were negatively correlated with PT and APTT, and positively correlated with FIB, D-D values and the levels of p50, p65, TAK1 and IκBα (4 p<0.05). CONCLUSIONS: MP-TF can lead to high blood coagulation in patients with breast cancer through the activation of NF-κB signaling pathway, which may become a new target for the intervention of the disease.