骨唾液酸蛋白基因沉默抑制乳腺癌细胞与骨基质粘附

Inhibitory effect of bone sialoprotein silencing on the adhesion ability of breast cancer cells to bone matrix

摘要:

旨在探索骨唾液酸蛋白 (Bone sialoprotein，BSP) 基因沉默对亲骨转移乳腺癌细胞 (MDA-MB-231BO) 与骨基质粘附能力的影响，为以BSP 为靶点的乳腺癌骨转移预防和靶向治疗提供实验依据。体外检测BSP 基因沉默对乳腺癌细胞与小鼠骨基质粘附能力的影响，MTS 法检测细胞增殖能力；扫描电镜观察骨片表面肿瘤细胞粘附情况和骨吸收状况；ELISA 法检测骨基质细胞粘附培养上清中TGF-β₁ 和RANKL 表达分泌量差异；左心室注射法构建裸鼠骨转移模

Abstract:

We performed this research mainly to explore the effect of bone sialoprotein (BSP) silence by siRNA on the adhesion ability to bone matrix of bone-seeking breast cancer cells (MDA-MB-231BO). Also we aimed to provide experimental data for prevention and treatment of breast cancer bone metastasis by targeting BSP. We explored the effects of BSP gene silence on characteristics of bone-seeking breast cancer cells: proliferation by MTS[3-(4,5-dimethylthiazol-2-yl)-5- (3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay, bone adhesion ability by a mouse bone adhesion model in vitro, morphology of the cells by SEM, and secretion of transforming growth factor-β₁ (TGF-β₁) and receptor activator of nuclear factor-kappa B ligand (RANKL) by ELISA kits. We performed intra-cardiac injection in nude mice to explore bone metastatic ability of different cell lines. The results showed that knockdown of BSP significantly inhibited the proliferation of MDA-MB-231BO cells and their adhesion to bone matrix. We also observed bone destruction caused by bone resorption around some adhering cells. The appearances of the cells changed in BSP gene silenced group, and the secretion of TGF-β₁ and RANKL decreased. The results showed BSP gene silence can partial inhibition bone metastasis of breast cancer cells in nude mice by X-ray assay and hematoxylin-eosin staining. Based on our research, siRNA-mediated BSP silencing can inhibit proliferation and adhesion to bone matrix of bone-seeking breast cancer cells and change their surface structure, thus inhibits their bone metastatic ability.