Vascular endothelial growth factor receptor-1 activation induces epithelial to mesenchmal transition in hepatocellular carcinoma cell line MHCC97-H / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To evaluate the mechanism of increased invasion and migration of hepatocellular carcinoma (HCC) cells induced by vascular endothelial growth factor receptor-1 (VEGFR-1) activation.</p><p><b>METHODS</b>Vascular endothelial growth factor-B (VEGF-B) was used to induce and stimulate hepatocellular carcinoma cell line MHCC97-H. Morphologic changes of MHCC97-H were investigated. The expression of E-cadherin and alpha-catenin (two epithelial markers) and Vimentin and N-cadherin (two mesenchymal markers) was detected by reverse transcriptase polymerase chain reaction (RT- PCR), western blotting, and immunofluorescence staining. Cell invasion and migration test was performed.</p><p><b>RESULTS</b>Treatment of MHCC97-H cells with VEGF-B led to morphologic changes characteristic of epithelial to mesenchymal transition (EMT), including loss of polarity, increased intercellular separation, and the presence of pseudopodia. Expression of the epithelial adhesion molecules, including E-cadherin and alpha-catenin, was decreased after VEGF-B treatment. Conversely, an increase in the expression of the mesenchymal cell markers, including N-cadherin and vimentin, was observed after VEGF-B treatment (P less than 0.05). VEGF-B-treated cells exhibited a change in E-cadherin from an organized, membrane-bound structure to a disorganized state in which it was noted to be dispersed throughout the cytoplasm. Pretreatment with VEGFR-1 blocking antibody 18F1 inhibited the change in localization of E-cadherin induced by VEGF-B treatment. The ability of invasion and migration of MHCC97-H was enhanced by VEGF-B reatment (P less alpha 0.05).</p><p><b>CONCLUSION</b>Increased invasion and migration of HCC cells induced by VEGFR-1 activation was mediated by epithelial to mesenchymal transition.</p>

The expression and significance of vascular endothelial growth factor and its receptors in hepatocellular carcinoma cell lines with various metastatic potentialities / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To examine whether or not vascular endothelial growth factor (VEGF) and its receptors were expressed in hepatocellular carcinoma cell lines with various metastatic potentialities.</p><p><b>METHODS</b>Reverse transcription-polymerase chain reaction (RT-PCR), enzyme linked immunosorbent assay (ELISA) and Western blot were employed to study the expressions of VEGFR-1, VEGFR-2, VEGF-A and VEGF-B in four hepatocellular carcinoma cell lines MHCC97-H, MHCC97-L, SMMC7721 and HepG-2 and one normal liver cell line L-02.</p><p><b>RESULTS</b>Three hepatocellular carcinoma cell lines (MHCC97-H, MHCC97-L, SMMC7721) expressed VEGFR-1 mRNA and their proteins. The expression level of VEGFR-1 in MHCC97-H was higher than that in MHCC97-L (P less than 0.05) and the expression level of VEGFR-1 in MHCC97-L was higher than that in SMMC7721 (P less than 0.05). No expression of VEGFR-1 was found in HepG-2 or L-02. All four hepatocellular carcinoma cell lines and the L-02 cell line expressed VEGFR-2 mRNA and the protein, as well as the VEGFR-1 ligands VEGF-A and VEGF-B. The expression level of VEGFR-2 in all tested hepatocellular carcinoma cell lines and normal liver cell line L-02 showed no significant differences (P more than 0.05).</p><p><b>CONCLUSION</b>VEGFR-1 was expressed in 4 hepatocellular carcinoma cell lines with various metastatic potentialities. The expression levels appeared to be positively correlated with the potentialities of metastasis of the hepatocellular carcinoma cell lines. VEGFR-1 may relate to the invasiveness and metastatic potential of the hepatocellular carcinoma.</p>