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Archive > Volume 23 Issue 5 > 2016,23(5):656-662. DOI:10.3872/j.issn.1007-385X.2016.05.012 Prev Next
Basic Research
Heparanase-mediated adhesion with vascular endothelial cell and trans-endothelial migration of hepatocellular carcinoma cells
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Abstract:
Objective:To explore the effect of heparanase (HPSE) on adhesion with vascular endothelial cell and trans-endothelial migration of hepatocellular carcinoma (HCC) cells. Methods:Human normal umbilical vein endothelial cell HUVEC-C line , hepatic LO-2 cells, and hepatic carcinoma HepG2, BEL-7402 and HCCLM3 cells were selected to use. HCC cells with high expression of HPSE were screened using Real-time PCR. Four RNA interference (RNAi) sequences of HPSE gene were designed, and the corresponding plasmids were constructed respectively, and the RNAi plasmids with the highest efficiency were screened out, which were transfected into HCC cells with high expression of HPSE. At the same time, blank control (BC) group, negative control (NC) group and untransfected HCC cell group were set up. Adhesion test was used to detect adhesion situation of HCC cell and HUVEC-C cell in each group and Transwell assay was performed to determine the trans-HUVEC-C cell migration of HCC cells. The HCC cells were stained with 0.25% Rose Bengal and observed under inverted microscope. After decolorization, D values of the HCC cells in each group were determined and compared using an enzyme-labeled instrument. Results: Expression level of HPSE mRNA in the HCCLM3 cells was significantly higher than those in hepatic LO-2, HepG-2 and Bel-7402 cells (\[5.72±0.62\] vs \[1.05±009\], \[2.65±0.31\] and \[3.43±0.58\], all P<0.01 ). Of the designed 4 RNAi plasmids of HPSE, siHPSE-3158 plasmid showed the best inhibitory effect on expression of HPSE gene (P<0.05). The adhesion rate of HCCLM3 cells and HUVEC-C cells in RNAi group was significantly lower than those in BC, NC and untransfected HCCLM3 cell groups (\[031±0.04\] vs \[0.46±0.06\], \[0.45±0.05\] and \[0.64±0.09\], all P<0.01). The trans-HUVEC-C cell migration rate of HCCLM3 cell in RNAi group was also significantly lower than those in BC, NC and untransfected HCCLM3 cell group (\[0.284±0.029\] vs \[0.41±0.04\], \[0.41±0.05\] and \[0.43±0.05\], all P<0.05). Conclusion: HPSE might be involved in mediating the adhesion of HCC cells and vascular endothelial cells as well as trans-endothelial cells migration, which could be a key factor in the adhesion to vascular endothelial cells and hematogenous metastasis of HCC cells.
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Project Supported:
Project supported by the National Natural Science Foundation of China（No.81272412）
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