Small RNA interference-mediated gene silencing of heparanase abolishes the invasion, metastasis and angiogenesis of gastric cancer cells

Jihua Dong,Guosong Jiang,Liduan Zheng,Qiangsong Tong,Hong Mei,Xiaohua Hou,Jiarui Pu

doi:10.1186/1471-2407-10-33

Jihua Dong, Guosong Jiang + Show 5 more

Open Access

https://doi.org/10.1186/1471-2407-10-33

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Abstract

BackgroundHeparanase facilitates the invasion and metastasis of cancer cells, and is over-expressed in many kinds of malignancies. Our studies indicated that heparanase was frequently expressed in advanced gastric cancers. The aim of this study is to determine whether silencing of heparanase expression can abolish the malignant characteristics of gastric cancer cells.MethodsThree heparanase-specific small interfering RNA (siRNAs) were designed, synthesized, and transfected into cultured gastric cancer cell line SGC-7901. Heparanase expression was measured by RT-PCR, real-time quantitative PCR and Western blot. Cell proliferation was detected by MTT colorimetry and colony formation assay. The in vitro invasion and metastasis of cancer cells were measured by cell adhesion assay, scratch assay and matrigel invasion assay. The angiogenesis capabilities of cancer cells were measured by tube formation of endothelial cells.ResultsTransfection of siRNA against 1496-1514 bp of encoding regions resulted in reduced expression of heparanase, which started at 24 hrs and lasted for 120 hrs post-transfection. The siRNA-mediated silencing of heparanase suppressed the cellular proliferation of SGC-7901 cells. In addition, the in vitro invasion and metastasis of cancer cells were attenuated after knock-down of heparanase. Moreover, transfection of heparanase-specific siRNA attenuated the in vitro angiogenesis of cancer cells in a dose-dependent manner.ConclusionsThese results demonstrated that gene silencing of heparanase can efficiently abolish the proliferation, invasion, metastasis and angiogenesis of human gastric cancer cells in vitro, suggesting that heparanase-specific siRNA is of potential values as a novel therapeutic agent for human gastric cancer.

Highlights

Heparanase facilitates the invasion and metastasis of cancer cells, and is over-expressed in many kinds of malignancies
Previous reports have shown that heparanase (HPA), an endo-hD-glucuronidase, has the ability to cleave the heparan sulfate chain of heparin sulfate proteoglycans (HSPGs), and is one of the key enzymes involved in the invasion and metastasis of malignant tumors [9]
We demonstrated that silencing of HPA expression attenuated the in vitro invasion, metastasis and angiogenesis capabilities of gastric cancer cells

Summary

Introduction

Heparanase facilitates the invasion and metastasis of cancer cells, and is over-expressed in many kinds of malignancies. The aim of this study is to determine whether silencing of heparanase expression can abolish the malignant characteristics of gastric cancer cells. Previous reports have shown that heparanase (HPA), an endo-hD-glucuronidase, has the ability to cleave the heparan sulfate chain of HSPGs, and is one of the key enzymes involved in the invasion and metastasis of malignant tumors [9]. It has been established that a significant correlation of HPA over-expression is coupled with increased metastatic potential and decreased survival rates of cancer [16]. These studies suggest that HPA is correlative with invasion and metastasis of cancer cells, and served as an important target of cancer therapy

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