Antisense Oligonucleotide Against Clusterin Regulates Human Hepatocellular Carcinoma Invasion Through Transcriptional Regulation of Matrix Metalloproteinase-2 and E-Cadherin

• Main Authors: Bomin Yan, Jun Liang, Xuelong Jiao, Kejun Zhang, Yan Wang, Dong Chen
• Format: Article
• Language: English
• Published: MDPI AG 2012-08-01
• Series: International Journal of Molecular Sciences
• Subjects: hepatocellular carcinoma; metastasis; clusterin
• Online Access: http://www.mdpi.com/1422-0067/13/8/10594

Secreted clusterin (sCLU) has been shown to be overexpressed in metastatic hepatocellular carcinoma (HCC) tissue, and its overexpression in HCC cells increases cell migration and the formation of liver metastatic tumor nodules <em>in vivo</em>. In this study, we tested the hypothesis that sCLU plays a role in the invasiveness of human HCC and may be associated with its metastatic spread. HCCLM3, a human hepatocellular carcinoma cell line, was transiently transfected with an antisense oligonucleotide (ASO) against sCLU (OGX-011). HepG2 liver hepatocellular cells were transiently transfected with the pc.DNA3.1-sCLU plasmid to overexpress sCLU, and subsequently evaluated for effects on invasion and the expression of molecules involved in invasion. We observed that suppression of the <em>sCLU</em> gene significantly reduced the invasive capability of the highly invasive HCCLM3 cells, and <em>vice versa</em> in the low invasive HepG2 cell line. The results revealed that knockdown of <em>sCLU</em> by <em>OGX-011</em> resulted in a significant increase in the expression of E-cadherin and a decrease in <em>matrix metalloproteinase-2</em> (<em>MMP-2</em>) gene transcription. Overexpression of sCLU<em> </em>by transfection with pc.DNA3.1-sCLU significantly decreased the expression of E-cadherin and increased <em>MMP-2</em> gene transcription. These data were further verified by reverse transcription-PCR and Western blot analysis. A significant reduction in MMP-2<em> </em>expression and an increase in E-cadherin expression in sCLU-knockdown HCCLM3 cells were observed, as well as a significant increase in MMP-2<em> </em>expression and a decrease in E-cadherin expression in HepG2 cells overexpressing sCLU. These data indicate a role for sCLU in augmenting MMP-2<em> </em>transcription and decreasing E-cadherin expression. Our data show the involvement of sCLU in human HCC invasion, and demonstrate that silencing <em>sCLU</em> gene expression inhibits the invasion of human HCC cells by inhibiting MMP-2<em> </em>expression and promoting E-cadherin expression. Thus, OGX-011 could be an effective therapeutic agent for HCC.