Zhang Yong, Wang Zhen-ning, Zhang Xue, Xu Hui-mian, Jiang Li, Luo Yang, Xing Li-li, Xu Mi-duo, Li Juan
Department of Oncosurgery, No.1 Hospital of China Medical University, Shenyang 110001, China.
Zhonghua Yi Xue Za Zhi. 2007 Jun 26;87(24):1717-20.
To investigate the inhibitory effect of siRNA on heparanase expression and invasion ability gastric cancer cells.
A heparanase mRNA-targeting double-stranded siRNA was designed with the bioinformatics technology. Human gastric cancer cells of the line SGC7901 were cultured and transfected with the siRNA of the concentrations of 5, 10, 20, and 40 nmol/L respectively. Forty-eight hours later RT-PCR and Western blotting were applied to detect the mRNA and protein expression of heparanase. Millicell chamber assay was performed to detect the invasion ability of the SGC7901 cells. Blank control group and negative control group were set.
The mRNA expression level of the cells transfected with the siRNA of the concentrations 20 nmol/L and 40 nmol/L were 0.207 +/- 0.095 and 0.200 +/- 0.085 respectively, both significantly lower than that of the control group (0.60 +/- 0.09, both P < 0.05). Western blotting showed that the protein expression of heparanase of the different siRNA subgroups were all decreased dose-dependently; and no heparanase band was seen in the 40 nmol/L subgroup. The invasion rate of the siRNA group was significantly lower than that of the control group with a mean inhibition rate of (61 +/- 36)%.
RNAi inhibits the expression of heparanase and the invasion ability of human gastric cancer cells. Heparanase may be a new target in treatment of gastric cancer's metastasis.
研究小干扰RNA（siRNA）对胃癌细胞乙酰肝素酶表达及侵袭能力的抑制作用。
运用生物信息学技术设计靶向乙酰肝素酶mRNA的双链siRNA。培养人胃癌SGC7901细胞系，分别用浓度为5、10、20和40 nmol/L的siRNA转染。48小时后，采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测乙酰肝素酶的mRNA和蛋白表达。进行微孔板侵袭实验检测SGC7901细胞的侵袭能力。设置空白对照组和阴性对照组。
用20 nmol/L和40 nmol/L siRNA转染的细胞，其mRNA表达水平分别为0.207±0.095和0.200±0.085，均显著低于对照组（0.60±0.09，P均<0.05）。蛋白质免疫印迹法显示，不同siRNA亚组的乙酰肝素酶蛋白表达均呈剂量依赖性降低；40 nmol/L亚组未见乙酰肝素酶条带。siRNA组的侵袭率显著低于对照组，平均抑制率为（6l±36）%。
RNA干扰抑制人胃癌细胞乙酰肝素酶的表达及侵袭能力。乙酰肝素酶可能是治疗胃癌转移的新靶点。
