抑制组织蛋白酶b表达对胃癌bgc-823细胞.docVIP

抑制组织蛋白酶B表达对胃癌BGC-823细胞黏附、 增殖和侵袭的影响 王涛1 ，李敏2，臧文巧 2， 赵国强2# （1河南中医学院第一附属医院血液肿瘤科，河南，郑州， 450000 2郑州大学基础医学院微生物学与免疫学教研室，河南，郑州，450052） 通讯作者：赵国强 Email: zhaogq@126.com??? 第一作者: 王涛. 硕士.主治医师. Tel Email: zangwenqiao@ [摘要] 目的：探讨抑制组织蛋白酶B表达对胃癌BGC-823细胞黏附、增殖和侵袭的影响。方法：构建针对CB的siRNA表达载体，脂质体LipofectAmineTM2000细胞株BGC-823；细胞黏附实验检测各组胃癌细胞的黏附能力；细胞侵袭性实验检测各组胃癌细胞的侵袭能力，细胞增殖实验检测各组胃癌细胞的增殖能力。空白对照组无关siRNA对照组沉默1组和沉默2组细胞中，CB mRNA的相对表达量显著p0.01）；CB蛋白表达被显著抑制纤维黏连蛋白和基质胶黏附的胃癌细胞数目穿透基质胶的胃癌细胞数显著（p0.0）在2d、3d、4d、5d细胞孔内的吸光度值显著减少，差异有显著性（P﹤0.05）。而在无关siRNA对照组和空白对照组之间，CB mRNA的相对表达量两组的黏附细胞数目两组穿透基质胶的细胞数无显著性差异（p0.05）结论设计针对CB基因的 siRNA片段构建的siRNA载体能够有效干扰胃癌BGC-823细胞中CB mRNA和蛋白的表达。抑制胃癌BGC-823细胞CB基因表达，可以降低胃癌细胞的生长速度、降低胃癌细胞与基质黏附能力、降低胃癌细胞侵袭和转移的能力。胃癌；组织蛋白酶B；增殖；侵袭；黏附adhesion 、invasion and metastasis Capabilities of gatric cancer cells BGC-823 WANG Tao1 ,, LI Min 2, ZANG Wenqiao2,ZHAO Guoqiang2 1Department of Hemato-tumor, The First Affiliated Hospital College Of TCM of Henan, Zhengzhou 450000, 2Department of Microbiology and Immunology, Medical college of Basic Science, Zhengzhou University, Zhengzhou 450052 [Abstract] AIM:To explore the effect of siRNA target gene CB on the adhesion 、invasion and metastasis Capabilities of gatric cancer cells BGC-823 . METHODS: Construct siRNA expression vector for CB using molecule clone and LipofectAmineTM2000 was used to transfect BGC-823, a kind of gastric tumor cell line. Fluorescent quantitative PCR and western blot were used to test the expression of CB in BGC-823 cells. Techniques of cell adhesion experiments, invasion assay and proliferation test were used to investigate the adhesion, invasion and proliferation of BGC-823 cells respectively. RESULTS: BGC-823 cells in silencing groups showed lower CB mRNA compared with unrelated siRNA and blank control groups; The number of BGC-823 cells adhering and penetrating to ECM and fibronectin decreased significantly in silencing group 1 and 2 compared with unrelated siRNA and blank control groups (p0.01). The absorbance in sil