ABSTRACT
Objective To study the mechanism of carboxypeptidase E ( CPE ) in promoting the migration of lymphocytes and their subsets through vascular endothelial cells. Methods CRISPR/Cas9 technology was used to prepare cpe gene-knockout MS1 (Cpe-/-MS1) cells. Adhesion ability of lymphocytes to MS1 and Cpe-/-MS1 cells was analyzed with adhesion assay. Expression of adhesion molecules on these cells were detected by RT-PCR and flow cytometry. Transwell model was used to compare the difference in the transmigration of lymphocytes and their subsets through MS1 and Cpe-/-MS1 cells. Results Cpe-/-MS1 cells were successfully obtained. Under the stimulation of TNF-α, the adhesion ability of lymphocytes to MS1 cells was much better than that of Cpe-/-MS1 cells. Moreover, adhesion molecules expressed on MS1 cells were significantly more than those on Cpe-/-MS1 cells. The percentages of lymphocytes and their sub-sets that transmigrated through MS1 cells were significantly higher than those through Cpe-/-MS1 cells. Con-clusion CPE involved in the adhesion of lymphocytes to vascular endothelial cells and the transmigration of them through vascular endothelial cells, which was of great significance for understanding the migration of lymphocytes across vascular endothelial cells to peripheral lymph nodes.
ABSTRACT
Objective Carboxypeptidase E ( CPE) is one of the most important carboxypeptidases in-volved in the biosynthesis of numerous peptide hormones and neurotransmitters and plays an important role in en -docrine regulation .A splice variant of CPE ( CPE-ΔN) has been detected and the mechanism of CPE -ΔN action in tumorigenesis has been studied in many different cancers .The aim of this study is to examine CPE -ΔN ex-pression in human colorectal cancer ( CRC) and to evaluate its possible application as a potential prognostic mark-er.Methods In the present study ,we have analyzed isoform expression of CPE -ΔN by qRT-PCR and West-ern blot in 219 CRC patients .Results Our results showed that CPE -ΔN was overexpressed in colorectal tumor tissue and that high CPE -ΔN mRNA expression was closely correlated with tumor differentiation ,pT classifica-tion,pN classification,tumor recurrence and lymph node metastasis .High CPE-ΔN expression was also identi-fied as an independent prognostic factor using a multivariate analysis (P=0.011).Conclusion Based on these results,we can conclude that CPE -ΔN expression might be a potential prognostic marker for CRC patients .