Effects of endothelial cells conditioned medium on the proliferation and invasion of hepatocellular carcinoma cells

Abstract

Objective To investigate the effects of vascular endothelial cells on the proliferation and invasion of hepatocellular carcinoma cells,and better understand the role of endothelial cells in the development of hepatocellular carcinoma (HCC).Methods The biological characteristics of Hep3B cells,such as proliferation,migration and invasion,were comparatively analyzed under HUVECs-derived conditioned medium (CM) stimulation and endothelial cell basal medium (EBM) stimulation,and its related signal molecules were detected by Western blotting.Data analysis was performed using SPSS 16.0 software.Results Cell Counting Kit-8 (CCK-8) test showed that the proliferation of Hep3 B cells treated with CM was significantly increased as compared with that of the control cells (P ＜ 0.01).The wound healing assay and cell migration assay demonstrated that the migration of Hep3B cells was obviously enhanced underCM stimulation [(33.65±1.22)％ vs.(57.85±2.20)％,P＜0.01 and 51.2±3.1 vs.28.4± 4.6,P ＜0.01],and invasion ability of Hep3B cells induced by CM was also improved (16.2 ± 2.3 vs.5.4 ± 1.5).The expressions of invasion/metastasis associated genes matrix metalloproteinase-2 (MMP-2),matrix metalloproteinase-9 (MMP-9),osteopontin (OPN) and cluster of differentiation (CD44) and the serine-threonine kinase (Akt) phosphorylation and Extracellular Regulated Protein kinases (ERK) phosphorylation level were all upregulated in the treated Hep3B with CM.Conclusion Vascular endothelial cell conditioned mediun containing some soluble cytokines and/or chemokines which may facilitate the proliferation,migration and invasion of HCC cells via paracrine pathway. Key words: Hepatocellular carcinoma ; Endothelial cell ; Paracrine ; Invasiveness and metastasis