Effects of silibinin on cell growth and invasive properties of a human hepatocellular carcinoma cell line, HepG-2, through inhibition of extracellular signal-regulated kinase 1/2 phosphorylation

The purpose of the current study is to evaluate the effect of silibinin on human hepatocellular carcinoma HepG-2 cells. Microculture tetrazolium test (MTT assay), Lactate dehydrogenase (LDH) release, Gelatin zymography, Griess reaction, Cell-based the extracelluar signal-regulated kinase (ERK) 1/2 phosphorylation assay and quantitative real-time RT-PCR were employed to appraise the effect of silibinin on cell proliferation, cytotoxicity, metastatic potential, nitric oxide (NO) production, ERK 1/2 phosphorylation and activation in HepG-2 cells. Silibinin inhibited cell proliferation, matrix metalloproteinase 2 enzymatic activity. NO production and ERK 1/2 phosphorylation in a dose-dependent manner without exerting any cytotoxicity effect. In addition, an expressive increase in mRNA levels of Raf kinase inhibitor protein (RKIP), sprouty-related protein 1 with EVH-1 domain (Spred-1), sprouty-related protein with EVH-1 domain 2 (Spred-2) coupled with a significant reduction in transcriptional levels of highly expressed in cancer (Hec1) and MMP-2 were observed. Altogether, these issues show for the first time that silibinin treatment could inhibit cell proliferation and invasive potential of HepG-2 cells through inhibition of ERK 1/2 cascade both directly (through suppression of ERK 1/2 phosphorylation) and indirectly (through up-regulation of RKIP, Spred-1 and Spred-2). In addition, cell growth and proliferation may be inhibited by silibinin through down-regulation of Hec1. (C) 2008 Elsevier B.V. All rights reserved.