Effects Of Claudin-1 SiRNA On The Dissociation And Invasion Ability Of Pancreatic Cancer Cells

ObjectiveTo observe the effects of silencing claudin-1gene by short interfering RNA(siRNA) on the Dissociation and invasion ability of pancreatic cancer cells.MethodsUsing claudin-1 specific siRNA transfected low metastatic pancreatic cancer cell line PC-1.The transfection efficiency of claudin-1 were confimed by RT-PCR semi-quantitative analysis, cells dissociated state used Papanicolaou 'S staining, the invasion ability was detemined by transwell chambers. The migration ability was detemined by Migration experiments and scratch assay.ResultsAfter claudin-1 gene-specific siRNA transfected 48 hours, mRNA expression level was significantly reduced compared with the negative control group decreased by 82.85%(GeneTools from SynGene gray value calculation), there was significant difference (P<0.01).Cell dissociation status:Papanicolaou'S staining, microscopy (200×) The cells were observed by microscopy (200×) with Papanicolaou'S staining, we found that pancreatic cancer cells in the negative control group showed island-like PC-1 clone cells, and siRNA transfected cells after 48 hours Solution status changes from the beginning were scattered in a single cell growth.Transwell invasion chamber analysis:The cells with the chamber containing Transwell Matrigel, incubated for 48hours, Papanicolaou'S staining cells were observed and counted 5 random view (200×) within the cells.then we found that the number of invasive cells in negative control group was 17.56±2.33, and claudin-1 siRNA transfected group was 33.67±2.35, the number of invasive cells were significantly increased, the difference was statistically significant (P=0.000, P=0.000).Transwell chamber analysis migration capabilities:The cells incubated for 24 hours, Papanicolaou'S staining cells were observed and counted 5 random view (200 x) within the cells, the number of migrating cells found in the negative control were 16.27±1.28, And claudin-1 siRNA transfection after 32.30±1.49, significantly increased the number of migrating cells, the difference was statistically significant (P=0.000, P=0.000).Experimental Analysis of migratory capacity scratch:scratch after the 24h, we found that siRNA group migration to the original injury area,but the control group was not obvious.ConclusionsAfer the siRNA transfected PC-1, the membrane cell junction claudin-1 expression was significantly reduced, resulting in dissociation of pancreatic cancer cells, and the transwell experiments demonstrated its invasion increased. Therefore, in the hamster pancreatic cancer cells, the expression of claudin-1 in pancreatic cancer cell are closely related to the invasion and metastasis, and a negative correlation.