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

Effect of siRNA target gene CB on adhesion,invasion and metastasis of gastric cancer cells BGC-823

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转染胃癌细胞株BGC-823。荧光定量RT-PCR和免疫印迹检测CB在各组胃癌细胞中的表达水平;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。结果与空白对照组和无关siRNA对照组相比,沉默1组和沉默2组细胞中,CB mRNA的相对表达量显著降低(P<0.01);CB蛋白表达被显著抑制;纤维粘连蛋白和基质胶黏附的胃癌细胞数目以及穿透基质胶的胃癌细胞数显著降低(P<0.05);并且在2 d、3 d、4 d及5 d细胞孔内的吸光度值亦显著减少,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以及两组穿透基质胶的细胞数都无显著性差异(P>0.05)。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。

【Objective】 To explore the effect of silencing CXCR4 by siRNA on adhesion,invasion and metastasis of gastric cancer cells BGC-823.【Methods】 siRNA expression vector for CB was constructed and then transfected into gastric cancer cells BGC-823 by LipofectAmineTM2000 RT-PCR and western blot were used to test the expression of CB in BGC-823 cells.Cell adhesion assay,invasion assay and proliferation test were used to investigate the adhesion,invasion and proliferation capabilities of BGC-823 cells respectively.【Results】 As compared with unrelated siRNA and blank control groups,the expression of CB mRNA in silencing group 1 and group 2 significantly decreased（P 0.01）;The average amount of cells adhering and penetrating to ECM and fibronectin was significantly decreased（P 0.05）.The absorbance significantly decreased at 2 d,3 d,4 d and 5 d（P 0.05）.There was no significant difference between unrelated siRNA and blank control groups（P 0.05）.【Conclusion】 Constructed siRNA vectors for CB could effectively decrease CB mRNA and protein expression in BGC-823 cells.The inhibition of cathepsin B expression could suppress growth,adhesion,invasion and metastasis of gastric tumor cells.