Effects of down-regulating expression of PDGF-D in human gastric carcinoma cells on the growth and function of co-cultured HUVECs
AIM: To study the effect of down-regulating expression of PDGF-D in human gastric carcinoma cell line SGC-7901 by RNA interference on the growth and function of co-cultured human umbilical vein endothelial cells ( HUVECs). METHODS:The short hairpinRNA (shRNA) eukaryotic expression vectors targeting PDGF-D were constructed and transfected into SGC-7901 cells. The changes of PDGF-D expression were detected by reverse transcriptase-polymerase chain reaction ( RT-PCR) and Western Blot. The concentration of VEGF in supemate was tested by enzyme linked immunosorbent assay ( EI.ISA). After the co-culture of SGC-7901 cells and HUVECs, the changes of cell growth, cell cycle, and angiogenic ability of HUVECs in vitro were detected by cell counting, flow cytometry, and matrigel in vitro HUVECs tube formation assay, respectively. RESULTS: The eukaryotic expression vectors of PDGF-D-shRNA1 and PDGF-D-shRNA2 could effectively inhibit PDGF-D expression in human gastric carcinoma cell line SGC-7901, and the latter was better(P < 0. 05 ). The concentration of VEGF in supemate decreased after RNA interference. The proliferation of HUVECs was inhibited after co-cultured with SGC-7901 cells, HUVECs could not form capillary tube-like structures. CONCLUSION: The eukaryotic expression vectors could effectively inhibit PDGF-D expression in human gastric carcinoma cell line SGC-7901, and inhibit the growth of HUVECs and the ability of capillary tube-like structure formation through decreasing VEGF expression.
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