RESUMO
OBJECTIVE: To investigate the correlation between Pokemon gene and cisplatin mechanism. METHODS: Human lung adenocarcinoma cells of the lines A549 and AGZY83-a, human lung squamous carcinoma cells of the line HE-99, and human giant cell lung cancer cells of the line 95D were cultured and cisplatin was added into the medium. Other lung cancer cells of the above mentioned lines were cultured in the medium without cisplatin and were used as control groups. RT-PCR and Western blotting were used to detect the mRNA and protein expression of Pokemon. RESULTS: Pokemon mRNA and protein were expressed highly in all the 4 cell lines. The Pokemon gene expression did not changed significantly after cisplatin treatment groups. There were not significant differences in the mRNA and protein expression of Pokemon among the 4 experiment groups and the control groups (all P > 0.05). CONCLUSION: Cisplatin has no effect on the Pokemon gene expression of the human lung cancer cells.
Assuntos
Cisplatino/farmacologia , Proteínas de Ligação a DNA/genética , Fatores de Transcrição/genética , Antineoplásicos/farmacologia , Western Blotting , Linhagem Celular Tumoral , Proteínas de Ligação a DNA/metabolismo , Resistencia a Medicamentos Antineoplásicos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Transcrição/metabolismoRESUMO
OBJECTIVE: To investigate the effect of Ras association domain family 1A (RASSF1A) gene, a new tumor suppressor gene (TSG), on tumorigenesis of human esophageal carcinoma cells. METHODS: pcDNA3.1 (+)-RASSF1A, a plasmid containing RASSF1A gene, and the blank plasmid pcDNA3.1 (+) were transfected into human esophageal carcinoma cells of the line EC9706. The expression of RASSF 1A protein was examined by Western blotting. The changes of cell cycle of stably-transfected cells were determined by flow cytometry (FCM), and the cellular proliferation was analyzed by MTT assay. Fifteen nude mice were randomly divided into 3 groups to be inoculated subcutaneously with EC9706 cells transfected with pcDNA3.1 (+)-RASSF1A, EC9706 cells transfected with pcDNA3.1 (+), and untransfected EC9706 cells respectively. Other 5 nude mice were used as controls. Four weeks later, the mice were killed to take out the carcinoma tissues. FCM was used to analyze the cell cycle. RESULTS: Western blotting showed that RASSF1A protein was expressed highly in the stably transfected cells. The cell viability and growing speed were decreased obviously in the cells expressing of RASSF1A (both P < 0.01); FCM showed that the proportion of cells at the G(1) phase of the EC9706 cells expressing RASSF1A was significantly higher than those in the blank plasmid group and untransfected group (both P < 0.01). The size of the EC9706 cells obtained from the nude mice inoculated with the EC9706 cells transfected with pcDNA3.1 (+)-RASSF1A was significantly smaller than those of the pcDNA3.1 (+) group and blank plasmid group (both P < 0.05). CONCLUSION: Expression of exogenous RASSF1A inhibits the progression of human esophageal carcinoma cells in vitro and in vivo. As a tumor suppressor gene, it plays an important role in origination, progression and metastasis of esophageal carcinoma.
Assuntos
Proliferação de Células , Neoplasias Esofágicas/genética , Proteínas Supressoras de Tumor/genética , Ensaios Antitumorais Modelo de Xenoenxerto , Animais , Ciclo Celular/genética , Ciclo Celular/fisiologia , Linhagem Celular Tumoral , Sobrevivência Celular/genética , Sobrevivência Celular/fisiologia , Neoplasias Esofágicas/patologia , Neoplasias Esofágicas/terapia , Feminino , Citometria de Fluxo , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Nus , Plasmídeos/genética , Transfecção , Proteínas Supressoras de Tumor/fisiologiaRESUMO
OBJECTIVE: To investigate the mRNA expression of the new tumor suppressor gene, RASSF1A, in esophageal squamous cell carcinoma and its biological value. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of RASSF1A in the tumor tissues, tissues near tumor, and normal tissues, all obtained during operation, from 66 esophageal squamous cell carcinoma patients. RESULTS: The deletion rates of RASSF1A mRNA expression were 42.4% (28/66), 15.2% (10/66), and 0 in the tumor tissues, tissues near tumor, and normal tissues respectively. The deletion rates of RASSF1A mRNA expression in patients with lymph node metastasis was 61.1%, significantly higher than that of the patients without lymph node metastasis (20.0%, chi(2) = 11.323, P < 0.01); The deletion rates of RASSF1A mRNA expression in the esophageal squamous cell carcinoma at the advanced stages (stages III - IV) was 61.5%, significantly higher than that in the esophageal squamous cell carcinoma at the early stages (stages I - II, 30.0%, chi(2) = 6.417, P < 0.01). The deletion rages of RASSF1A mRNA in the highly, moderately, and lowly differentiation tumors were 38.5% (10/26), 38.5% (10/26), and 57.1% (8/14) respectively, However, there was no significant association between the differentiation degree of tumor and the grade the deletion rages of RASSF1A mRNA (chi(2) = 1.576, P = 0.455). CONCLUSION: The deletion of the mRNA expression of RASSF1A, a tumor suppressor gene, may play an important role in the tumorigenesis and influences the prognosis of esophageal squamous cell carcinoma.
