| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
Fiscal Year 2012: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000)
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| Research Abstract |
This is a challenging study to explore biomarkers on tumor cell-derived nanovesicles (exosomes) and to investigate their potential as innovative diagnostic markers for early detection and molecular characterization of cancer, and for decision making in cancer treatment. Here, we performed a focused proteome analysis for the proteins of exosomes, which are recently revealed to be membrane vesicles secreted from various cells including tumor cells. At first, tumor cell-derived exosomes were collected by ultracentrifugation method from cell culture supernatant. The finally collected products were confirmed to be purified exosomes by morphology of electron microscopy, particle size by dynamic light scattering and expression of tetraspanins, which were markers of exosomes, by western blotting. Then, proteins were extracted from the purified exosomes and analyzed the expressed proteins by mass spectrometry. These experiments revealed that various tumor-related membrane proteins were detectable in tumor-derived exosomes. In the future, we will test the feasibility of exoxome-based in vitro diagnosis by isolating the tumor-derived exoxomes from the peripheral blood of patients and profiling tumor-related proteins of them.
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