Quantitative analysis of the secretome of TGF-beta signaling-deficient mammary fibroblasts.
Transforming growth factor beta (TGF-beta) is a master regulator of autocrine and paracrine signaling pathways between a tumor and its microenvironment. Decreased expression of TGF-beta type II receptor (TbetaRII) in stromal cells is associated with increased tumor metastasis and shorter patient survival. In this study, SILAC quantitative proteomics was used to identify differentially externalized proteins in the conditioned media from the mammary fibroblasts with or without intact TbetaRII. Over 1000 proteins were identified and their relative differential levels were quantified. Immunoassays were used to further validate identification and quantification of the proteomic results. Differential expression was detected for various extracellular proteins, including proteases and their inhibitors, growth factors, cytokines, and extracellular matrix proteins. CXCL10, a cytokine found to be up-regulated in the TbetaRII knockout mammary fibroblasts, is shown to directly stimulate breast tumor cell proliferation and migration. Overall, this study revealed hundreds of specific extracellular protein changes modulated by deletion of TbetaRII in mammary fibroblasts, which may play important roles in the tumor microenvironment. These results warrant further investigation into the effects of inhibiting the TGF-beta signaling pathway in fibroblasts because systemic inhibition of TGF-beta signaling pathways is being considered as a potential cancer therapy.
Xu
Baogang J
BJ
Department of Cancer Biology, Vanderbilt University, Nashville, TN, USA. baogang.j.xu@vanderbilt.edu
Yan
Wenwei
W
Jovanovic
Bojana
B
An
Angel Q
AQ
Cheng
Nikki
N
Aakre
Mary E
ME
Yi
Yajun
Y
Eng
Jimmy
J
Link
Andrew J
AJ
Moses
Harold L
HL
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D049109:Cell Proliferation; D005347:Fibroblasts / Q000737:chemistry* / Q000378:metabolism; D008321:Mammary Glands, Animal / Q000737:chemistry* / Q000166:cytology / Q000378:metabolism; D051379:Mice; D017346:Protein Serine-Threonine Kinases / Q000172:deficiency / Q000378:metabolism; D020543:Proteome / Q000032:analysis* / Q000378:metabolism; D000077294:Receptor, Transforming Growth Factor-beta Type II; D018125:Receptors, Transforming Growth Factor beta / Q000172:deficiency / Q000378:metabolism; D015398:Signal Transduction; D016212:Transforming Growth Factor beta / Q000378:metabolism*
D020543:Proteome; D018125:Receptors, Transforming Growth Factor beta; D016212:Transforming Growth Factor beta; D017346:Protein Serine-Threonine Kinases; D000077294:Receptor, Transforming Growth Factor-beta Type II
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