| Objective(1)To detect the expression of chemokine receptors 7 (CXCR7) in different degree of malignancy human hepatocellular carcinoma (HCC) cell lines.(2)To explore the role of CXCL12/CXCR7 axis in the proliferation and migration of hepatocellular carcinoma cells.Methods(1)The expression of CXCR7 protein and micro-RNA in hepatocellular carcinoma cell lines MHCC97-H (with high metastatic tumor) and PLC/PRF/5 (with low metastatic tumor) was measured by Western blot analysis and RT-PCR analysis. By ICC ( immunocytochemistry) and FACS ( Fluorescence Activating Cell Sorter ), the expression of CXCR7 in two hepatocellular carcinoma cell lines were determined at the level of membrane protein. By LSCM (laser scanning confocal microscope), the expression of CXCR7 in hepatocellular carcinoma cell line MHCC97-H cell was determined at the subcellular localization.(2)After the addition of CXCL12 and CXCR7 blocker is CCX771, the changes of cell proliferation were observed by Alamarblue and cellular migration assay were observed by Transwell system. To investigate the effect of CXCL12/CXCR7 axis on hepatocellular carcinoma cells chemotaxis. The experiment design of control group, chemokine group and inhibition group.Results1. Western blotting, RT-PCR, ICC and FACS showed that MHCC97-H cells expressed higher CXCR7 levels than PLC/PRF/5 cells.2. The growth rate and migration in MHCC97-H cells was increased after CXCL12 treatment (p<0.01), and significantly decreased after the addition of CCX771 (p<0.01). However, PLC/PRF/5 cells had no response to CXCL12 (p>0.05).Conclusion CXCL12/CXCR7 axis may play an important role in high metastatic potential HCC cell lines proliferation and migration, and CXCR7 blocker CCX771 can efficiently suppress this effect. |
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