Influence of osteopontin expression on the metastatic growth of CC531 rat colorectal carcinoma cells in rat liver.

Abstract

Development of hepatic metastasis is responsible for most of colorectal cancer-related deaths. Osteopontin (OPN) is a small integrin-binding N-linked glycoprotein, which plays a crucial role in the formation of hepatic metastasis. This study aimed to suppress Opn expression by an antisense-oligonucleotide (ASO(Opn)) to decrease liver metastasis in vivo. The effect of ASO(Opn) was investigated in vitro in CC531(lacZ) colorectal cancer cells in comparison to sense (SO) or nonsense (NSO) oligomers, by determining mRNA and protein expression levels, as well as cell survival. For in vivo treatment, CC531(lacZ) cells were intraportally inoculated into rats to compare the effects of ASO, SO and NSO oligomers, following prolonged subcutaneous administration by osmotic mini-pumps. The resulting CC531(lacZ) tumor cell load in the liver was measured by a β-galactosidase assay. Proliferation of CC531(lacZ) cells in vitro was significantly decreased after ASO(Opn) and SO treatment (P<0.001). Liver metastasis development was reduced as long as ASO(Opn) was administered, but this effect was rapidly blunted following the end of the ASO(Opn) administration. In contrast, administration of the SO resulted in a tumor load reduction, which surprisingly surpassed the ASO(Opn) effect in vivo in terms of a long-lasting metastasis suppression, which was accompanied with increased survival of the animals. Administration of the ASO(Opn) in rats was effective in decreasing their liver metastasis. The short-lived effect might be extended by modifications suited to increase the ASOs' half-life. In addition, there was a superior anti-metastatic effect caused by the SO, which has not been reported previously.