Total exosomal protein levels in the serum were analyzed using BCA assay, and exosomal BP1 protein levels were determined using Western blot. The hypothesis that exosomal BP1 protein may be related to the development of breast cancer was investigated with clinical serum samples purchased from Capital Bioscience. Levels of BP1 protein were determined in cell extracts (CE) and conditioned media (CM) using Western Blot analysis. In cell line experiments, fluorescent immunohistochemistry was used to detect the location of BP1 protein. Materials and Methods: Exosomes were isolated using the commercially available Exosome Precipitation Solution. Therefore, we hypothesized that BP1 protein may be packed in exosomes, and analysis of exosomal BP1 protein could provide a novel biomarker for diagnosis or prognosis of breast cancer. In addition, researchers found that exosomal proteins have unique characteristics compared to traditional biomarkers for carcinoma diagnosis and prognosis. Exosomes are nano-sized membrane-bound vesicles released by various cells and play important roles in intercellular communication. BP1 overexpression was found to stimulate known oncogenes including BCL2 and c-Myc. Moreover, high BP1 levels were associated with tumor cell survival, breast cancer aggressiveness and metastasis. Previous work showed BP1 mRNA was activated in 80% of invasive ductal breast (IDC) tumors, where 100% of estrogen receptor (ER) negative tumor tissues and 73% of ER positive tumor tissues were BP1 positive. Background: Beta protein 1 (BP1), discovered in our lab, is an isoform protein of the DLX4 gene that is a member of the homeobox gene family.
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