Gut-derived serotonin induced by depression promotes breast cancer bone metastasis through the RUNX2/PTHrP/RANKL pathway in mice

Abstract

Abstract. Breast cancer metastasizes to the bone in a majority of patients with advanced disease resulting in bone destruction. The underlying mechanisms are complex, and both processes are controlled by an interaction between locally and systemically derived signals. Clinically, breast cancer patients with depression have a higher risk of bone metastasis, yet the etiology and mechanisms are yet to be elucidated. MDA-MB-231 breast cancer cells were used to establish a bone metastasis model by using intracardiac injection in nude mice. Chronic mild stress (CMS) was chosen as a model of depression in mice before and after inoculation of the cells. Knockdown of the RUNX-2 gene was performed by transfection of the cells with shRNA silencing vectors against human RUNX-2. A co-culture system was used to test the effect of the MDA-MB-231 cells on osteoclasts and osteoblasts. RT-PCR and western blotting were used to test gene and protein expression, respectively. We confirmed that depression induced bone metastasis by promoting osteoclast activity while inhibiting osteoblast differentiation. Free serotonin led to an increase in the expression of RUNX2 in breast cancer cells (MDA-MB-231), which directly inhibited osteoblast differentiation and stimulated osteoclast differentiation by the PTHrP/RANKL pathway, which caused bone destruction and formed osteolytic bone lesions. Additionally, the interaction between depression and breast cancer cells was interrupted by LP533401 or RUNX2 knockdown. In conclusion, depression promotes breast cancer bone metastasis partly through increasing levels of gut-derived serotonin. Activation of RUNX2 in breast cancer cells by circulating serotonin appears to dissociate coupling between osteoblasts and osteoclasts, suggesting that the suppression of gut-derived serotonin decreases the rate of breast cancer bone metastasis induced by depression. IntroductionBreast cancer is one of the most common cancers worldwide and tends to metastasize to bone, resulting in osteolysis and skeletal-related events (pain and fracture) in patients. The mechanisms underlying this metastasis are complex and involve both particular characteristics of the breast cancer cells and the bone matrix (soil and seed concept). During metastasis, breast cancer cells possess certain properties that enable them to grow in bone, and the bone matrix provides a suitable microenvironment which facilitates the growth of breast cancer cells. The bone microenvironment facilitates dynamic bone resorption and bone formation which maintains the balance of bone mass. Bone remodeling consists of both resorption by osteoclasts (1-5) and new bone formation by osteoblasts (3,5). The two processes of remodeling are coupled temporally and spatially (1,3,5,6) and are controlled by an interplay between locally and systemically derived signals, such as mechanical strain, growth factors, hormones and other molecules (1,5,6). Breast cancer cells disrupt this normal physiological process and generate factors that directly or indirectly induce th...

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