Prognostic and predictive impact of stroma cells defined by PDGFRb expression in early breast cancer: results from the randomized SweBCG91RT trial.

PURPOSE: Predictive biomarkers are needed to aid the individualization of radiotherapy (RT) in breast cancer. Cancer-associated fibroblasts have been implicated in tumor radioresistance and can be identified by platelet-derived growth factor receptor-beta (PDGFRb). This study aims to analyze how PDGFRb expression affects RT benefit in a large randomized RT trial. METHODS: PDGFRb was assessed by immunohistochemistry on tissue microarrays from 989 tumors of the SweBCG91RT trial, which enrolled lymph node-negative, stage I/IIA breast cancer patients randomized to RT after breast-conserving surgery. Outcomes were analyzed at 10 years for ipsilateral breast tumor recurrence (IBTR) and any recurrence and 15 years for breast cancer specific death (BCSD). RESULTS: PDGFRb expression correlated with estrogen receptor negativity and younger age. An increased risk for any recurrence was noted in univariable analysis for the medium (HR 1.58, CI 95% 1.11-2.23, p = 0.011) or PDGFRb high group (1.49, 1.06-2.10, p = 0.021) compared to the low group. No differences in IBTR or BCSD risk were detected. RT benefit regarding IBTR risk was significant in the PDGFRb low (0.29, 0.12-0.67, p = 0.004) and medium (0.31, 0.16-0.59, p < 0.001) groups but not the PDGFRb high group (0.64, 0.36-1.11, p = 0.110) in multivariable analysis. Likewise, risk reduction for any recurrence was less pronounced in the PDGFRb high group. No significant interaction between RT and PDGFRb-score could be detected. CONCLUSION: A higher PDGFRb-score conferred an increased risk of any recurrence, which partly can be explained by its association with estrogen receptor negativity and young age. Reduced RT benefit was noted among patients with high PDGFRb, however without significant interaction.

Endostatin suppresses IGF-II-mediated signaling and invasion of human extravillous trophoblasts.

Endostatin, a biological active fragment of the extracellular matrix protein collagen XVIII, is known to interfere with cellular motility in the context of pathological angiogenesis. However, the physiological role of endostatin remains largely elusive. Recent evidence suggested that the inhibitor is produced in human decidual cells of early pregnancy, indicating that endostatin could be involved in diverse reproductive processes, such as implantation and/or placental differentiation. To gain more insights into the role of endostatin, we here analyzed its effects on trophoblast motility, proliferation, and signaling using purified primary trophoblasts, first-trimester villous explant cultures, and trophoblastic SGHPL-5 cells. In vitro Transwell assays demonstrated that purified endostatin inhibited both basal and IGF-II-induced migration and invasion as well as outgrowth from villous explant cultures. In contrast, basal and IGF-II-stimulated proliferation was unaffected upon addition of the inhibitor. Analyses of IGF-II-associated downstream signaling events showed that endostatin interfered with activation of various signaling kinases such as ERK1/2, protein kinase B (Akt)/mammalian target of rapamycin/p70 S6 kinase, and focal adhesion kinase. Furthermore, virus-mediated, stable gene silencing of Akt1 in SGHPL-5 cells using a micro-RNA-adapted short hairpin RNA-expressing plasmid revealed that endostatin-mediated inhibition of IGF-II-induced Akt phosphorylation was critically dependent on the expression of the particular isoform. In conclusion, the data suggest that endostatin could be a physiological inhibitor of IGF-II-dependent trophoblast cell motility by suppressing focal adhesion kinase/Akt/mammalian target of rapamycin/p70 S6 kinase signaling.