The Identification Of Metastasis-associated Protein ATP5B And Its Localization In PC3M Cells

Prostate cancer is the most common malignancy in the male urinarysystem. In2010, statistics show that the incidence of prostate cancerranks first in the United States. In recent years, the incidence of prostatecancer is rising. About80%of prostate cancer patients will eventuallyappear the situation of distant metastases. And the transfer mechanism ofprostate cancer is unclear.In early experiments, we get the peptides B04which can bind a highmetastatic prostate cancer cell lines. What’s more, through in vitro and invivo experiments, we confirm its ability which inhibits prostate cancercell lines with high metastatic adhesion, invasion and metastasis. Sincethe peptide B04has the ability to inhibit metastasis, therefore, we believethat peptide B04receptor involved in the biological behavior of invasionand metastasis PC3M cells. To find and confirm the B04peptide receptor,we use the proteomic approach to finish the purification and identificationof PC3M cell membrane protein. Furthermore, the distribution of ATP5Bhas been studied in PC3M cells.Results:1. By Western blot, we analysis membrane and plasma proteins ofPC3M cell. Experimental results show thatB04receptor protein is the membrane proteins of PC3M cell.2. Using the peptide B04coupled to Sepharose4B, we finished affinitychromatography of PC3M cell membrane proteins. Further, weanalysis the eluate by polyacrylamide gel electrophoresis andcoomassie blue staining. The result shows that the position of strip ofthe purified protein sample is consist with results of Western blot.3. The samples were analyzed by mass spectrometry bands. The resultsthat we obtained the identity information of131proteins. Throughsearching the literature and summering preliminary experimentalresults, we have made up a conclusion that ATP5B is one of the B04receptor.4. We analysis the expression and localization of ATP5B byimmunohistochemistry and immunofluorescence. Experimentalresults show that ATP5B expressed on the cell membrane andmitochondria in PC3M cells.To sum up, ATP5B is the receptor of the peptide B04in prostatecancer cell lines PC3M. It’s located in the cell membrane andmitochondria in PC3M cells. Due to B04has the ability to inhibitmetastasis in PC3M cells. We think that ATP5B is a protein whichassociates with metastasis in PC3M cells. Peptide-specific B04is through ATP5B play its action. The expression on the cell membrane may be thekey to play suppression functions.