The Role Of SNX5 In The Proliferation Of The Breast Cancer

Objective: Sorting nexins(SNXs) containing SNX-PX domains(a subtype of phosphate domain family) of a class of the outer peripheral proteins. As part Retromer complex, SNXs assist Retromer composite material transport from the Golgi apparatus to the membrane system, so SNXs family has associated with membrane transport and material decomposition. As SNX5 SNXs an important family in papillary thyroid cancer is highly expressed in lung, breast, colon, liver and ovarian cancer with low expression. Tip SNX5 may play a role in the development of malignant tumors. This study focused on SNX5 expression in breast cancer cell proliferation, and to further explore the connection SNX5 expression and regulation of cell proliferation between SNX5 elucidate the role of cell cycle pathway might play.Content: Analysis SNX5 in tumor tissue specimens in cell localization; SNX5 construction of eukaryotic expression vector; Construction SNX5 high expression of tumor cell lines; the use of small interfering RNA silencing construct SNX5 tumor cell lines clarify the relationship between the cycle-related genes SNX5; After flow cytometry overexpression and silencing SNX5 to analyze the changes in the cell cycle.Method: In this study, firstly the prognosis of breast cancer based on gene expression profiles online prediction website(Gene expression-based outcome for breast cancer Online, GOBO) forecast SNX5 expression and prognosis in breast cancer patients, and immunohistochemical techniques tumor tissue the SNX5 staining, observed under a microscope by cell coloring situation in the cytoplasm,the nucleus. Furthermore, we Experimental Research SNX5 specific mechanism of action in vitro, using the PCR technique to amplify the target gene SNX5. And by connecting with the eukaryotic expression vector was constructed SNX5 eukaryotic expression vector. Expression plasmids using non-liposomal Fu GENE? HD Transfection Reagent-mediated transfection SNX5 over-expression plasmid, that is to build SNX5 high-expressing tumor cell lines. We purchased SNX5 small interfering RNA, and constructed by transfection of cell lines SNX5 silence.Application of CCK-8 method and colony, and its relationship with cell proliferation SNX5 EDU detected between the application and flow cytometry SNX5 influence on cell cycle. Double-Thymidine by law, the cells were synchronized in the G1 / S phase and the cell cycle distribution by flow cytometry and Western Blot method and cell cycle checkpoint protein expression. Finally, the relationship between the use of cell cycle-related gene assay kit SNX5 cycle related genes and between genes.Through our study is the first to explore the SNX5 breast cancer cell proliferation,research SNX5 role in the regulation of cell cycle progression in the play, and explore the relationship between SNX5 cell cycle-related genes.Results: We found that, SNX5 protein is highly expressed in cancer tissues and low expression in cancerous peripheral tissues. In MCF-7 and MDA-MB-231 breast cancer cell lines, after applying si RNA silencing SNX5, cell proliferation was significantly inhibited, and after SNX5 expression in MCF-7 high, cell proliferation increased significantly, indicating SNX5 promote breast cancer cells proliferation. In MDA-MB-231 and MCF-7 breast cancer cell lines, silence SNX5 promote cancer cell arrest in G1 / S phase or G2 / M phase, and after overexpression SNX5 promote cells to enter S phase, indicating SNX5 associated with the cell cycle. To further confirm SNX5 by double thymidine block method Hela cells release in G1 / S phase,Flow cytometry showed that the expression of silence SNX5 confirmed cancer cells arrest in G2 / M phase. Cell cycle-related genes kit display, silence SNX5 expression,P15 and P16 gene expression increased, suggesting that P15 and P16 gene expression in silence SNX5 possible arrest in G2 / M phase associated with cancer cells.Conclusion: SNX5 is closely related to the proliferation of breast cancer, and it maybe control the cancer cell proliferation by the cell cycle.