Hypoxia-inducible factor–dependent breast cancer–mesenchymal stem cell bidirectional signaling promotes metastasis
Pallavi Chaturvedi, … , Andre Levchenko, Gregg L. Semenza
Pallavi Chaturvedi, … , Andre Levchenko, Gregg L. Semenza
Published December 17, 2012
Citation Information: J Clin Invest. 2013;123(1):189-205. https://doi.org/10.1172/JCI64993.
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Research Article

Hypoxia-inducible factor–dependent breast cancer–mesenchymal stem cell bidirectional signaling promotes metastasis

Abstract

Metastasis involves critical interactions between cancer and stromal cells. Intratumoral hypoxia promotes metastasis through activation of hypoxia-inducible factors (HIFs). We demonstrate that HIFs mediate paracrine signaling between breast cancer cells (BCCs) and mesenchymal stem cells (MSCs) to promote metastasis. In a mouse orthotopic implantation model, MSCs were recruited to primary breast tumors and promoted BCC metastasis to LNs and lungs in a HIF-dependent manner. Coculture of MSCs with BCCs augmented HIF activity in BCCs. Additionally, coculture induced expression of the chemokine CXCL10 in MSCs and the cognate receptor CXCR3 in BCCs, which was augmented by hypoxia. CXCR3 expression was blocked in cocultures treated with neutralizing antibody against CXCL10. Conversely, CXCL10 expression was blocked in MSCs cocultured with BCCs that did not express CXCR3 or HIFs. MSC coculture did not enhance the metastasis of HIF-deficient BCCs. BCCs and MSCs expressed placental growth factor (PGF) and its cognate receptor VEGFR1, respectively, in a HIF-dependent manner, and CXCL10 expression by MSCs was dependent on PGF expression by BCCs. PGF promoted metastasis of BCCs and also facilitated homing of MSCs to tumors. Thus, HIFs mediate complex and bidirectional paracrine signaling between BCCs and MSCs that stimulates breast cancer metastasis.

Authors

Pallavi Chaturvedi, Daniele M. Gilkes, Carmen Chak Lui Wong, Kshitiz, Weibo Luo, Huafeng Zhang, Hong Wei, Naoharu Takano, Luana Schito, Andre Levchenko, Gregg L. Semenza

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Figure 10

HIF and PGF expressed by BCCs are required for MSC migration and homing.

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HIF and PGF expressed by BCCs are required for MSC migration and homing....
(A and B) Migration of MSCs in response to CM isolated from NTC, shPGF-1, and shPGF-2 MDA-231 subclones cultured at 20% or 1% O2. MSCs were seeded on the top of the Boyden chamber, and the number of cells that migrated through the filter in response to CM in the lower chamber was counted under light microscopy after staining with crystal violet. Data were normalized to CM isolated from NTC cells at 20% O2. **P < 0.001 vs. 20% NTC; ##P < 0.005 vs. 1% NTC. Scale bar: 200 μm. (C) 1 × 106 NTC, shPGF-1, or shPGF-2 cells were implanted into the MFP of SCID mice. Recruitment of MSCs to the primary tumor was analyzed by qPCR for SRY (mean ± SEM; n = 5). *P < 0.05 vs. NTC, Student’s t test. (D) Representative images acquired by time-lapse photomicroscopy of labeled MDA-231 BCCs (green) and MSCs (red) cocultured in a LiveAssay 2-chamber device coated with fibronectin. Scale bar: 200 μm. (E and F) Migration of MSCs (E) and EV or DKD BCCs (F) after 12 hours of coculture. *P < 0.05, **P < 0.01 vs. 20% EV; ##P < 0.01 vs. 1% EV. (G and H) Migration of MSCs (G) and NTC or shPGF-1 BCCs (H) after 12 hours of coculture. *P < 0.05, **P < 0.01 vs. 20% NTC; #P < 0.05, ##P < 0.01 vs. 1% NTC.