ABSTRACT
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.
Subject(s)
Humans , Cadherins , Cell Movement , Genetics , Neoplasm Invasiveness , Neoplasm Metastasis , Stomach Neoplasms , Genetics , Metabolism , Pathology , Transfection , Tumor Cells, Cultured , Twist-Related Protein 1 , Genetics , VimentinABSTRACT
OBJECTIVE@#To determine the effect of EphA2 protein on the expression of vascular endothelial growth factor (VEGF) and matrix metalloproteinase 9 (MMP9) proteins in HCT116 cells.@*METHODS@#High expression of EphA2 protein in HCT116 cells was confirmed by Western blot. HCT116 cells were transfected with EphA2 antisense oligonucleotide. The expression of the transfection efficiency was analyzed by Western blot. VEGF proteins in the cell supernatants were detected by enzyme linked immunosorbent assay(ELISA), and the expressions of MMP9 in cell supernatants were examined by gelatin zymography.@*RESULTS@#EphA2 antisense oligonucleotide suppressed the expression of VEGF and MMP9 proteins in HCT116 cells.@*CONCLUSION@#EphA2 could decrease the invasion and metastasis of HCT116 cells by suppressing the expression of VEGF and MMP9.
Subject(s)
Humans , HCT116 Cells , Matrix Metalloproteinase 9 , Metabolism , Oligonucleotides, Antisense , Receptor, EphA2 , Genetics , Metabolism , Vascular Endothelial Growth Factor A , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effect of DPC4 gene expression on the growth of colon cancer cells and its mechanism.</p><p><b>METHODS</b>Expression plasmid pcDNA3.1-DPC4 was constructed and transfected into the colon cancer cell line SW620 by use of lipofectamine gene transfer technique. DPC4 protein expression was detected by Western blot and immunocytochemistry. The effect of DPC4 gene on the growth of SW620 cells was monitored by population doubling time (PDT) and cloning efficiency. The influence of DPC4 expression on p21WAF1 transcription was investigated by RT-PCR to detect p21WAF1 mRNA.</p><p><b>RESULTS</b>Successful expression of DPC4 protein was detected in the transfected SW620 cells. Compared with the control cells, PDT (74 h) of the DPC4 expressing cells was prolonged and the cloning efficiency (21%) decreased. In addition, the mRNA level of p21(WAF1) in DPC4 transfected cells was increased.</p><p><b>CONCLUSIONS</b>Overexpression of DPC4 gene inhibits the growth of colon cancer in vitro, and induction of p21(WAF1) expression may be an important functional aspect of DPC4.</p>