RESUMO
beta-Catenin, which is frequently overexpressed in a variety of human cancers including esophageal cancer, mediates cancer cell proliferation and tumor growth. In the present study, we used a human U6 promoter-driven DNA-template approach to induce short hairpin RNA (shRNA)-triggered RNA interference to silence beta-catenin gene expression in human esophageal squamous cell carcinoma cell line Eca-109, and then evaluated its effects on the proliferation and growth of tumor cells in vitro and in nude mice. beta-Catenin expression levels decreased markedly in Eca-109 cells transfected with a plasmid expressing shRNA for beta-catenin. Downregulation of beta-catenin was concomitantly accompanied by reduction of cyclin D1, colony formation, and growth inhibition of Eca-109 cells in vitro. The mechanism appears to be the G0/G1 phase arrest but not induction of apoptosis. In vivo, treatment of Eca-109 cells with beta-catenin shRNA greatly impeded tumor growth in nude mice. We conclude that plasmid vector-mediated beta-catenin RNA interference holds great promise as a novel treatment on human esophageal cancer with beta-catenin overexpression.
Assuntos
Interferência de RNA/fisiologia , RNA Interferente Pequeno/genética , beta Catenina/genética , Animais , Caspase 3/metabolismo , Linhagem Celular Tumoral , Proliferação de Células , Ciclina D1/fisiologia , Fase G1/genética , Humanos , Camundongos , Camundongos Nus , Fase de Repouso do Ciclo Celular/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução de Sinais , Células Tumorais Cultivadas , Ensaios Antitumorais Modelo de Xenoenxerto , beta Catenina/metabolismoRESUMO
OBJECTIVE: To explore the effect of Twist gene on the migration and invasion of human gastric carcinoma cells. METHODS: MKN28 cells, a human gastric carcinoma cell line, were transfected with PcDNA3.1-Twist plasmid by lipofectamine transfecting technique. The transfected cells were selected with geneticin. Expressions of Twist,ecadherin and vimentin protein were detected by Western blot in cells transfected Twist gene. Matrigel invision chambers were performed to analyse the cell migration and invasion. RESULTS: MKN28 cells transfected with PcDNA3.1-Twist plasmid showed stronger intracellular expression of Twist protein than MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The expression of ecadherin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly decreased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without the transfection. However, The expression of vimentin protein in MKN28 cells transfected with PcDNA3.1-Twist plasmid was significantly increased compared with that in MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. The migration and invasion ability of Twist+ - MKN28 cells were stronger than that of MKN28 cells transfected with PcDNA3.1 and MKN28 cells without transfection. CONCLUSION: Twist gene may promote the migration and invasion ability of gastric carcinoma cells through epithelial mesenchymal transition.
Assuntos
Movimento Celular/genética , Neoplasias Gástricas/genética , Neoplasias Gástricas/patologia , Proteína 1 Relacionada a Twist/genética , Caderinas/biossíntese , Humanos , Invasividade Neoplásica , Metástase Neoplásica , Neoplasias Gástricas/metabolismo , Transfecção , Células Tumorais Cultivadas , Proteína 1 Relacionada a Twist/biossíntese , Vimentina/biossínteseRESUMO
OBJECTIVE: To explore the relationship between the expressions of PTEN and the metastasis of the gallbladder cancer. METHODS: The expression of PTEN and nm23 were detected by immunohistochemical staining in 32 cases of gallbladder cancer with metastasis and the staining intensity was scored semi-quantitatively, compared with the cases without metastasis. RESULTS: The intensity score of PTEN and nm23 in gallbladder cancer with metastasis was 8.9947+/-4.5590 and 10.2003+/-3.9031, respectively, which was lower than that in those without metastasis (12.9433+/-4.7618 and 15.8436+/-5.6917 respectively, P < 0.01 ). The expression of PTEN was correlative with that of nm23 ( Pearson = 0.370, P < 0.05). CONCLUSION: The lower expressions of PTEN and nm23 are related to the metastasis of gallbladder cancer.
Assuntos
Neoplasias da Vesícula Biliar/metabolismo , Neoplasias Hepáticas/secundário , Núcleosídeo-Difosfato Quinase/biossíntese , PTEN Fosfo-Hidrolase/biossíntese , Feminino , Humanos , Neoplasias Hepáticas/metabolismo , Metástase Linfática , Masculino , Nucleosídeo NM23 Difosfato Quinases , Núcleosídeo-Difosfato Quinase/genética , PTEN Fosfo-Hidrolase/genética , Proteínas Supressoras de Tumor/biossíntese , Proteínas Supressoras de Tumor/genéticaRESUMO
To detect the expression of miRNA-214 in human gastric cancer cell lines of BGC823, MKN45 and SGC7901, and to identify the effect of miRNA-214 on cell cycle and apoptosis of these cells. Expression of miRNA-214 in human normal gastric mucosal cell line GES-1 and human gastric cancer cell lines was detected by real-time reverse-transcription polymerase chain reaction. Antisense-miRNA-214 oligonucleotides were transfected transiently into gastric cancer cell lines to down-regulate the expression of miRNA-214. The cell cycle and apoptosis were studied by flow cytometry assay. PTEN, one of the target genes of miRNA-214 was detected by using of immunocytochemistry and Western blotting. MiRNA-214 was overexpressed in gastric cancer cell lines of BGC823, MKN45 and SGC7901 compared with normal gastric mucosal cell line GES-1. Antisense-miRNA-214 oligonucleotides significantly down-regulated the expression of miRNA-214, and increased the portion of G1-phase and decreased the portion of S-phase in BGC823 and MKN45 cells. The immunocytochemistry test and Western blotting analysis showed that the down-regulation of miRNA-214 could significantly up-regulate the expression of PTEN in BGC823 and MKN45 cells. MiRNA-214 is overexpressed in human gastric cancer cell lines of BGC823, MKN45 and SGC7901. The down-regulation of miRNA-214 could induce a G1 cell cycle arrest in them, the up-regulation of PTEN maybe one of the mechanism.
Assuntos
Pontos de Checagem do Ciclo Celular/genética , Fase G1/genética , Regulação Neoplásica da Expressão Gênica , MicroRNAs/genética , PTEN Fosfo-Hidrolase/genética , Neoplasias Gástricas/genética , Apoptose/genética , Western Blotting , Linhagem Celular , Linhagem Celular Tumoral , Regulação para Baixo , Humanos , MicroRNAs/biossíntese , Oligonucleotídeos Antissenso/genética , PTEN Fosfo-Hidrolase/biossíntese , Reação em Cadeia da Polimerase em Tempo Real/métodos , Fase S/genética , Neoplasias Gástricas/metabolismo , Transfecção , Regulação para CimaRESUMO
OBJECTIVE: To study the pathological basis of metastatic brain tumors misdiagnosed with (MRI). METHODS: Seven cases were analyzed histopathologically and immunohistochemically, compared with the MRI image. RESULTS: All the tumors were single large noduls. Hemorrhage and necrosis of the tumors were seen in all the cases. Cystformation of the tumor, glial cell proliferation and vascular proliferation of peritumoral brain tissues were seen in most cases. CONCLUSION: The secondary changes of metastatic brain tumors and the reactive changes of peritumoral brain tissues are the major causes of MRI misdiagnosis.