Department of Surgery, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China.
BMC Cancer. 2010 Feb 5;10:33. doi: 10.1186/1471-2407-10-33.
Heparanase 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.
Three 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.
Transfection 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.
These 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.
肝素酶促进癌细胞的侵袭和转移，在许多恶性肿瘤中过度表达。我们的研究表明，肝素酶在晚期胃癌中经常表达。本研究旨在确定沉默肝素酶表达是否可以消除胃癌细胞的恶性特征。
设计、合成了三个肝素酶特异性小干扰 RNA（siRNA），并转染到培养的胃癌细胞系 SGC-7901 中。通过 RT-PCR、实时定量 PCR 和 Western blot 检测肝素酶的表达。通过 MTT 比色法和集落形成实验检测细胞增殖。通过细胞黏附实验、划痕实验和基质胶侵袭实验检测癌细胞的体外侵袭和转移。通过内皮细胞管形成实验检测癌细胞的血管生成能力。
转染针对编码区 1496-1514 bp 的 siRNA 导致肝素酶表达减少，这种减少始于转染后 24 小时，并持续 120 小时。肝素酶介导的沉默抑制了 SGC-7901 细胞的细胞增殖。此外，敲低肝素酶后，癌细胞的体外侵袭和转移能力减弱。此外，肝素酶特异性 siRNA 的转染以剂量依赖的方式减弱了癌细胞的体外血管生成。
这些结果表明，肝素酶基因沉默可以有效地消除人胃癌细胞在体外的增殖、侵袭、转移和血管生成，表明肝素酶特异性 siRNA作为一种新型治疗人胃癌的药物具有潜在价值。
