Objective: To explore the effects of up-regulated expression of C-Src kinase-binding protein (CBP) on biological functions and cell ultrastructure in Jurkat cells.
Methods: We constructed the CBP-EGFP viral vector and established a stably transfected Jurkat cell line with it. Cell growing status was observed under a light microscope. The transfection efficiency and the location of CBP were examined with confocal microscopy. The complexity of cell organelles was observed by electron microscopy. Apoptotic cells were detected by flow cytometry. The expressions of C-Src kinase (CSK) and Vav oncoprotein in CBP signaling pathway were measured by real-time quantitative PCR (qRT-PCR).
Results: We found more pyknotic cells and less homogeneity in the transfected Jurkat cells under light microscope. Confocal microscopy revealed that cell transfection efficiency was up to 100% and CBP was located on the cell membrane. Moreover, there were more complex organelles in CBP-EGFP transfected Jurkat cells with a lot of mitochondrions and the lysosome-like clumps accumulation. Detection of apoptosis showed that CBP-EGFP transfected Jurkat cells had more necrotic cells compared with the negative group. Finally, qRT-PCR indicated the expression of CSK increased and Vav decreased in the CBP-EGFP transfected group.
Conclusion: Up-regulated expression of CBP in Jurkat cells could reduce cell homogeneity and promote cell apoptosis.