Expression data from breast cancer bone metastasis [MDA-MB 231 scp1833]. Mus musculus

The goal of this study was to evaluate genes that are differentially expressed in the bone stroma when breast cancer metastasis are present. We focused our attention on bone stroma cells to understand how the dissemination and growth of tumor cells can impact on the bone environment. Moreover, we aimed to identify potential targets to inhibit the cross talk between cancer cells and tumor microenvironment in bone metastasis. MDA-MB 231 scp1833 breast cancer cell line were engineered to express luciferase and GFP, and injected in NOD-SCID mice by the intracardiac route to obtain bone metastasis. Tumor growth was monitored in-vivo by bioluminescence for 2 weeks. Endothelial cells and osteoblasts were isolated from the long bones of tumor-bearing and tumor-free mice to identify genes differentially expressed. Overall design: Bone marrow single cell suspensions from tumor-free or tumor-bearing mice (n=6-10 samples) were prepared by grinding bones in a mortar followed by enzymatic digestion of the bone powder for 1 hour at 37°C with 1 mg/ml collagenase, 1 mg/ml dispase and 50KU/ml DNAse. Stroma cells and hematopoietic fractions were enriched by discontinuous percoll density gradient separation (1.065 g/L and 1.115 g/L). Remaining red blood cells were lysed and the single cell population was stained to isolate two different stroma cells by FACS: osteoblasts (OBctr) (OBtum), were identified as GFP-TR119-CD45-SCA1-CD31-CD51+, and endothelial cells (ECctr) (ECtum) were identified as GFP-TR119-CD45-SCA1+CD31+.

本研究旨在评估存在乳腺癌转移时骨基质中差异表达的基因。我们聚焦骨基质细胞，以探究肿瘤细胞的播散与生长如何影响骨微环境。此外，本研究还旨在筛选可抑制骨转移中癌细胞与肿瘤微环境间交叉互作的潜在靶点。将MDA-MB 231 scp1833乳腺癌细胞株改造为表达荧光素酶（luciferase）与绿色荧光蛋白（Green Fluorescent Protein，GFP），通过心内注射途径接种至NOD-SCID小鼠体内以诱导骨转移。采用生物发光成像法在体监测肿瘤生长，时长共计2周。从荷瘤与无瘤小鼠的长骨中分离内皮细胞与成骨细胞，以鉴定差异表达的基因。总体实验设计：通过将骨骼在研钵中研磨，并于37℃下使用1 mg/ml胶原酶、1 mg/ml分散酶及50KU/ml脱氧核糖核酸酶对骨粉进行1小时酶解，制备无瘤或荷瘤小鼠的骨髓单细胞悬液（样本量n=6-10）。通过不连续Percoll密度梯度离心（1.065 g/L与1.115 g/L）富集基质细胞与造血细胞组分。裂解剩余红细胞后，对单细胞群体进行染色，通过荧光激活细胞分选术（Fluorescence Activated Cell Sorting，FACS）分离两种不同的基质细胞：成骨细胞（OBctr、OBtum），其鉴定标记为GFP⁻TR119⁻CD45⁻SCA1⁻CD31⁻CD51⁺；内皮细胞（ECctr、ECtum），其鉴定标记为GFP⁻TR119⁻CD45⁻SCA1⁺CD31⁺。