Molloy Alan P, Martin Fiachra T, Dwyer Roisin M, Griffin Tomas P, Murphy Mary, Barry Frank P, O'Brien Timothy, Kerin Michael J
Department of Surgery, National University of Ireland Galway, Galway, Ireland.
Int J Cancer. 2009 Jan 15;124(2):326-32. doi: 10.1002/ijc.23939.
Over 70% of patients with advanced breast cancer will develop bone metastases for which there is no cure. Mesenchymal Stem Cells (MSCs) and their derivative osteoblasts are subpopulations of cells within the bone marrow environment, postulated as potential interacting targets for disseminating cancer cells because of their ability to secrete a range of chemokines. This study aimed to investigate chemokine secretion throughout MSC differentiation into osteoblasts and their effect on the breast cancer cells. Primary MSCs and osteoblast progenitors were cultured in appropriate conditions to induce differentiation into mature osteoblasts. Chemokines secreted throughout differentiation were detected using ChemiArray and ELISA. Migration of breast cancer cells in response to the bone-derived cells was quantified using Transwell inserts. Breast cancer cells were cocultured with MSCs, retrieved using magnetic beads, and changes in CCL2 expression were analyzed. MSCs secreted a range of factors including IL-6, TIMP-1 and CCL2, the range and level of which changed throughout differentiation. CCL2 secretion by MSCs increased significantly above control cells as they differentiated into mature osteoblasts (p<0.05). The bone-derived cells stimulated migration of breast cancer cells, and this was inhibited (21-50%) in the presence of a CCL2 antibody. CCL2 gene expression in breast cancer cells was upregulated following direct coculture with MSCs. The varying levels of chemokines secreted throughout MSC differentiation may play an important role in supporting tumor cell homing and progression. These results further highlight the distinct effect MSCs have on breast cancer cells and their potential importance in supporting development of metastases.
超过70%的晚期乳腺癌患者会发生骨转移，而目前尚无治愈方法。间充质干细胞（MSCs）及其衍生的成骨细胞是骨髓环境中的细胞亚群，由于它们能够分泌一系列趋化因子，被认为是播散癌细胞潜在的相互作用靶点。本研究旨在调查间充质干细胞分化为成骨细胞过程中趋化因子的分泌情况及其对乳腺癌细胞的影响。将原代间充质干细胞和成骨祖细胞在适当条件下培养，以诱导分化为成熟的成骨细胞。使用化学发光阵列和酶联免疫吸附测定法检测分化过程中分泌的趋化因子。使用Transwell小室定量乳腺癌细胞对骨源细胞的迁移反应。将乳腺癌细胞与间充质干细胞共培养，用磁珠回收，分析CCL2表达的变化。间充质干细胞分泌一系列因子，包括白细胞介素-6、基质金属蛋白酶组织抑制因子-1和CCL2，其种类和水平在分化过程中发生变化。随着间充质干细胞分化为成熟的成骨细胞，其CCL2分泌显著高于对照细胞（p<0.05）。骨源细胞刺激乳腺癌细胞迁移，而在存在CCL2抗体的情况下，这种迁移受到抑制（21%-50%）。与间充质干细胞直接共培养后，乳腺癌细胞中CCL2基因表达上调。间充质干细胞分化过程中分泌的不同水平趋化因子可能在支持肿瘤细胞归巢和进展中发挥重要作用。这些结果进一步突出了间充质干细胞对乳腺癌细胞的独特作用及其在支持转移发展中的潜在重要性。
