Identification of potential tumor differentiation factor (TDF) receptor from steroid-responsive and steroid-resistant breast cancer cells.
作者： Izabela Sokolowska , Alisa G. Woods , Mary Ann Gawinowicz , Urmi Roy , Costel C. Darie
关键词: Molecular biology 、 Blot 、 Hsp70 、 Cellular differentiation 、 HeLa 、 Receptor 、 Heat shock protein 、 Cell 、 Biology 、 Cell type
摘要: Tumor differentiation factor (TDF) is a recently discovered protein, produced by the pituitary gland and secreted into bloodstream. TDF TDF-P1, 20-amino acid peptide selected from open reading frame of TDF, induce in human breast prostate cancer cells but not other cells. protein has no identified site action or receptor, its mechanism unknown. Here, we used TDF-P1 to purify identify potential receptor (TDF-R) candidates MCF7 steroid-responsive non-breast HeLa cancerous using affinity purification chromatography (AP), mass spectrometry (MS). We four candidate proteins 70-kDa heat shock (HSP70) family Experiments did any TDF-R candidates. AP MS experiments were validated Western blotting (WB). additionally looked for steroid-resistant BT-549 dermal fibroblasts (HDF-a) WB. interacts with HDF-a Immunofluorescence (IF) GRP78, candidate, at cell surface MCF7, cells, IF HSP70 demonstrated labeling on all types. These results point toward GRP78 as strong suggest that exclusively through steroid-independent pathway.
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