| Pancreatic cancer is known to be an extremely lethal neoplasm, one of the reasons being that pancreatic cancer itself has an extremely high potential of invasion-metastasis. In our previous study, two pancreatic cancer cell lines with a different potential for invasion-metastasis, PC-1 with a low potential and PC-1.0 with a high potential of invasion-metastasis after intrapancreatic transplantation, were established in a Syrian golden hamster. In our previous study, Differential expression of candidate genes Nup107 and claudin23 were indentified by cDNA microarray analysis.The nuclear pore complex (NPC) is a massive,multiprotin structure responsible for traffic between the nucleus and cytoplasm, it plays a critical role,not only in cell survival,but also in other cellular events such as differentiation. It's structure and function change is correlated to tumorigenesis and development. Nup107 is a component of the nuclear pore complex, a important nucleoporin.In the current study,the expression of Nup107 were increased in spongiocytoma cell.But it has no report about pancreatic cancer, and the mechanism remains incompletely understood.claudins are essential component of the intercellular tight junction, its are multigene family. Today, the claudin superfamily consists of 24 members in eukaryotes. the genetic expression change of claudin may induce endothelial cell and epithelial cell breakdown,and they are involved in many disease. The claudin family member may play an important role in tumorigenesis and development,also in invasion and metastasis. In the current study,the expression of claudin23 were decreased in intestinal type gastric cancer cell. But it has no report about signal transduction pathway, and the mechanism remains incompletely understood. In our previous study, the mitogen-activated protein kinase (MAPK) signal transduction pathway y plays a central role in pancreatic cancer progression. The special pathway align on epidermal growth factor receptor (EGFR)/mitogen-activated protein kinase kinase (MEK)/extracellular signal-regulated kinase (ERK). But the relationship between Nup107 or claudin23 and MAPK signal transduction pathway in pancreatic cancer is still unclear.In this study,intends to claudin23 by immunoocytochemistry and Nup107 through the expression of RNAi technology in the pancreatic cancer cell lines Preliminary study Nup107 and claudin23 in the pathogenesis of pancreatic cancer in the role in order to investigate the occurrence and development of pancreatic cancer relationshipObjectiveTo study the role of Nup107 and claudin23 in the pathogenesis of pancreatic cancer and molecule mechanisms; To provide a new targer to bring a new method of gene therapy for pancreatic cancer.MethodsUsing RT-PCR and Western-blot observed the mRNA and protein expression of Nup107 in PC-1 with a low potential and PC-1.0 with a high potential of invasion-metastasis. With Lipofectamine method Nup107 siRNA transfected into pancreatic cancer cells. Using RT-PCR and Western-blot observed the mRNA and protein expression of Nup107 and MEK in the PC-1.0 cells after transfection 48h and the flow cytometry, CCK-8, Papanicolaou'staining and a Matrix gel coated Tanswell plate were used separately to detect the cell cycle, changes in proliferation,cell dissociation and invasion ability. Using RT-PCR and Western-blot observed the mRNA and protein expression of Nup107 in the stable transfection MEKl-shRNA and MEK2-shRNA PC-1.0 cells. Using RT-PCR, Western-blot and immunoocytochemistry observed the mRNA and protein expression and distribution of claudin 23 ResultsThe mRNA and protein expression of Nup107 in PC-1.0 with a high potential of invasion-metastasis are higher than PC-1 with a low potential.Nup107 siRNA transfection group compared with the control group cells, Nup107's mRNA, protein expression was significantly reduced (P<0.01). cells'proliferation was inhibited after transfection transfected,cells increased the proportion of G1 phase, S phase,G2 phase cells decreased and cell dissociation and invasion ability were also restrained. Nup107's mRNA and protein expression was significantly reduced (P<0.01) in the stable transfection MEKl-shRNA PC-1.0 cells, but MEK2-shRNA PC-1.0 cells have no significant changes (P>0.05). The mRNA and protein expression of claudin23 in PC-1.0 with a high potential of invasion-metastasis are lower than PC-1 with a low potential (P<0.05). After incubation with dissocation factor conditioned medium(DF-CM) of PC-1.0 cells, plasma membrane distribution of claudin23 was obviously disrupted, and expressions of claudin23 decreased in PC-1 cells. claudin23 's mRNA and protein expression was significantly increased (P<0.05) in the stable transfection MEK1-shRNA PC-1.0 cells, but MEK2-shRNA PC-1.0 cells have no significant changes (P>0.05).ConclusionsNup107 has a key role of cell cycle, cell proliferation,cell dissociation and invasion ability in pancreatic cancer. Nup107 has important effect in the occurrence and development of pancreatic cancer cells. The effect of cell cycle and cell proliferation were through the cellular transduction MAPK/MEK1 pathway, The effect of cell dissociation and invasion ability were through the cellular transduction MAPK/MEK2 pathway. The effect of cell cycle, cell proliferation,cell dissociation and invasion ability were regulated the expression of Nup107 through the cellular transduction MAPK/MEK1/2 pathway. Arragement of expression and distribution of claudin23 is closely related to cell dissocation status in pancreatic cancer cell through MEK2 activation.
|
PDF Full Text Request