Small RNA interference-mediated gene silencing of heparanase abolishes the invasion, metastasis and angiogenesis of gastric cancer cells.
Small RNA interference-mediated gene silencing of heparanase abolishes the invasion, metastasis and angiogenesis of gastric cancer cells.
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小RNA干扰介导的乙酰肝素酶基因沉默消除胃癌细胞的侵袭、转移和血管生成
DOI：
10.1186/1471-2407-10-33
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发表时间：
2010-02-05
期刊：
影响因子：
3.8
通讯作者：
Tong Q
中科院分区：
文献类型：
--
作者：
Zheng L;Jiang G;Mei H;Pu J;Dong J;Hou X;Tong Q
背景乙酰肝素酶促进癌细胞的侵袭和转移，在多种恶性肿瘤中过度表达。我们的研究表明，乙酰肝素酶在进展期胃癌中频繁表达。本研究的目的是确定是否沉默乙酰肝素酶的表达可以取消胃癌细胞的恶性特征。MethodsThree乙酰肝素酶特异性小干扰RNA（siRNA）的设计，合成，并转染到培养的胃癌细胞株SGC-7901。采用RT-PCR、实时荧光定量PCR和Western blot检测乙酰肝素酶的表达。MTT比色法和集落形成实验检测细胞增殖情况。采用细胞粘附实验、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能力。结果针对1496-1514 bp编码区的siRNA转染导致乙酰肝素酶的表达降低，这种表达在转染后24小时开始，持续120小时。siRNA介导的乙酰肝素酶沉默抑制了SGC-7901细胞的增殖。此外，敲低乙酰肝素酶后，癌细胞的体外侵袭和转移能力减弱。结论乙酰肝素酶特异性siRNA能有效抑制胃癌细胞的增殖、侵袭、转移和血管生成，具有潜在的治疗胃癌的价值。
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. Thein vitroinvasion 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, thein vitroinvasion and metastasis of cancer cells were attenuated after knock-down of heparanase. Moreover, transfection of heparanase-specific siRNA attenuated thein vitroangiogenesis 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 cellsin vitro, suggesting that heparanase-specific siRNA is of potential values as a novel therapeutic agent for human gastric cancer.
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影响因子：
7.3
作者：
Karoli, T;Liu, LG;Ferro, V
通讯作者：
Ferro, V
影响因子：
10.3
作者：
Edovitsky, E;Elkin, M;Vlodavsky, I
通讯作者：
Vlodavsky, I
DOI：
10.1016/j.bbrc.2004.11.027
发表时间：
2005-01-14
影响因子：
3.1
作者：
Deng, H;Guo, RF;Lu, YY
通讯作者：
Lu, YY
影响因子：
8.4
作者：
Ikeguchi, M;Hirooka, Y;Kaibara, N
通讯作者：
Kaibara, N
克隆哺乳动物肝素酶--肿瘤侵袭和转移过程中的一种重要酶
影响因子：
82.9
作者：
Hulett, MD;Freeman, C;Parish, CR
通讯作者：
Parish, CR