Xiao Qun Qin

Cloning of a novel protein interacting with BRS-3 and its effects in wound repair of bronchial epithelial cells.

Bombesin receptor subtype 3 (BRS-3), the orphan bombesin receptor, may play a role in the regulation of stress responses in lung and airway epithelia. Bombesin receptor activated protein (BRAP )is a novel protein we found in our previous study which interacts with BRS-3. This study was designed to observe the subcellular location and wound repair function of BRAP in human bronchial epithelial cells (HBECs). BRAP ORF was amplified by RT-PCR and ligated to pEGFP-C1 vector, and then the recombinant plasmid pEGFP-C1-BRAP was transfected into Hela cells. The location of BRAP protein was observed by laser confocal microscope, and the expression of it was analyzed by Western-blot. At the same time,we built the recombinant plasmid pcDNA3.1(+)-BRAP, transfected it into HBECs and observed its impact on cell cycle and wound repair of HBECs. The results showed that BRAP locates in membrane and cytoplasm and increases significantly in transfected cells. Flow cytometry results demonstrated that the recombinant plasmid increases S phase plus G2 phase of cell cycle by 25%. Microscopic video analysis system showed that the repair index of wounded HBECs increases by 20% through stable expression of BRAP. The present study demonstrated that BRAP locates in the membrane and cytoplasm, suggesting that this protein is a cytoplasm protein, which promotes cell cycle and wound repair of HBECs.

Down‐regulation of integrin beta‐4 on human bronchial epithelial cells leads to inhibition of proliferation, wound repair and restricted anti oxidative damage

labeled proteins isolated from the normal human sperm and separated on the same 2-D gel along with a Cy2-labeled mixture of all samples as an internal standard. For the other gel, we labeled the sample reversed. Over 61 protein spots were analyzed in each paired normal/abnormal comparison, and using DIGE technology with the mixed-sample internal standard, 36 protein spots were identified by ms/ms as showing significant changes (paired t-test, p<0.05) in the level of expression between normal and round-headed. 9 proteins were up-regulated and 27 proteins were down-regulated in round-headed. These proteins seem to play important roles in a variety of pathways including spermatogenesis, cell signaling, cell skeleton and metabolism.