Differentially quantitative proteome of MHCC97H after Rab27B knock-down:a preliminary study / 军事医学

ABSTRACT

Objective To discover the vital role of Rab27B in tumor cells and its potential molecular mechanism by means of quantitative proteomics analysis of Rab27B knockdown in MHCC97H.Methods The expression of Rab27B in MHCC97H cells was knocked down by the combination of Tet-on advanced inducible expression system and RNA interference technology.Then, proteins extracted from the cells were identified by LC-MS/MS system after FASP digestion and iTRAQ 4-plex labeling. Finally, the properties of differentially expressed proteins, including the subcellular localizations, biological processes and molecular functions, were analyzed by the bioinformatics method.Results There were 448 differentially expressed proteins (|Ratio|>1.21, P<0.05) identified in MHCC97H cells after Rab27B knock-down.The expression levels of 229 or 219 proteins were positively or negatively correlated with Rab27B, respectively. These differentially expressed proteins were mainly involved in vesicle transport, macromolecule localization, cellular response to stimulus.Furthermore, there were 26 differentially expressed proteins participating in 8 tumor-related signal pathways, eleven of which were in the focal adhesion signal pathway.Conclusion The analysis of quantitative proteomics in Rab27B-knockdown MHCC97H cell line by iTRAQ suggests that Rab27B not only has an impact on the exosomal secretion of tumor cells, but also regulates master proteins in signal pathways involved in cell proliferation and migration.