Co-culture of hepatocellular carcinoma cells and human umbilical endothelial cells damaged by SU11274

Abstract

Mesenchymal-epithelial transition factor (c-Met) is a receptor that binds to the hepatocyte growth factor and is upregulated in hepatocellular carcinoma (HCC). The anti-tumor effects of (3Z)-N-(3-chlorophenyl)-3-({3,5-dimethyl-4-[(4- methyl-piperazin-1-yl)carbonyl]-1H-pyrrol-2-yl}methylene)-N-me- thyl-2-oxo-2,3-dihydro-1H-indole-5-sulfonamide (SU11274), a c-Met inhibitor, were investigated in the present study. HCC cells (HLE, HLF, PLC/PRL/5, Hep3B, Huh-6 and HepG2) and human umbilical vein endothelial cells (HUVECs) were used. Quantitative polymerase chain reaction was performed to detect the expression level of c-Met in HCC and HUVECs, and cyclin D1 in HCC. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-car-boxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tet razolium inner salt assay was performed to assess the proliferation of the HCC cells and HUVECs cultured with SU11274. Co-culture of HLF or PLC/PRL/5 cells and HUVECs was established as an in vitro model of HCC tissues. The expression levels of c-Met in HLE, HLF, PLC/PRL/5, Hep3B, Huh-6 and HepG2, adult healthy liver and HUVECs were 4.43+/-0.50, 1.61+/-0.18, 3.70+/-0.08, 0.81+/-0.18, 6.60+/-1.29, 1.06+/-0.35, 1.00+/-0.09 and 88.8+/-17.3 (mean +/- standard deviation), respectively. SU11274 (30 muM) suppressed the proliferation of HLF, PLC/PRL/5 and HUVECs to 11.0+/-9.4, 46.5+/-30.7 and 29.4+/-5.0%, respectively. SU11274 (30 muM) decreased the expression levels of cyclin D1 in HLF and PLC/PRL/5 cells to 45.1+/-11.6 and 30.1+/-10.3%, respectively. SU11274, at a concentration of 30 muM damaged the morphology of the co-cultures of HLF or PLC/PRL/5 cells with HUVECs and all the cells died. c-Met is highly expressed in HUVECs and HCC cells, but not in Hep3B. At a 30-muM concentration, SU11274 suppresses the proliferation of HLF, PLC/PRL/5 and HUVECs. SU11274 (30 muM) damages the co-cultures of HLF or PLC/PRL/5 cells with HUVECs.