| Background and Objective:Pancreatic cancer(PanCa)is a kind of common type malignant tumor in digestive system with a high case fatality rate.Invasion and metastasis is a common phenomenon of pancreatic cancer and is associated with local recurrence and pain.Nitric oxide(NO)regulated the cell growth,invasion and other behavior.Pancreatic cancer neural invasion(PNI)is a major way of the invasion and metastasis,and is also a special way for metastasis.By observing the expression of NO related proteins in clinical pathological specimens in PC,we explore the relationship between PNI and NO related proteins.Through the intervention of NO donor in PC,we explore the influence of NO on cell proliferation,migration,and invasion related proteins.Methods:(1)Using the immunohistochemical technique to test the expression of Ecadherin,iNOS,NQO1 and LC-3 in PC tissues,we analyzed the correlation between the factors with clinicopathologic characteristics and PNI.(2)We observed the influence of proliferation and apoptosis in different concentrations of DETA/NO by MTT test.(3)We observed the effect of DETA/NO on cell migration and invasion by wound healing and transwell test.(4)RT-PCR detected the changes of the cell associated protein mRNA level after DETA/NO intervention in PC cells.(5)The influence of DETA/NO impact on the expression of autophagy related protein expression was via Western blot.Results:(1)The expression rate of E-cadherin,iNOS,NQO1,and LC-3 were 68.9%,63.9%,55.7%,57.4% in PC respectively,E-cadherin was associated with LC-3,LC-3 was associated with local infiltration,and the expression of E-cadherin,LC-3 expression were significantly associated with PNI.(2)Low concentration DETA/NO can promote cell proliferation in PC,while high concentrations can inhibt cell proliferation.(3)The ability of migration and invasion was enhanced by the application of low concentration DETA/NO on PC cell lines.(4)The m RNA expression of i NOS,TNF,MMP-2 and MMP-9 were up regulated by the intervention of 50 μM DETA/NO-3 in Bx PC-3 cell lines.(5)The expression of LC-3 began to rise after 1h intervention of 100 uM DETA/NO,and it return to normal at 8h,but there was no obvious difference between NQO1 and Beclin-1.Conclusion:NO metabolism related proteins are closely related to the biological characteristics of PC,and it plays an important role in the progress of PC.Low concentration of NO can increased the ability of cell migration and invasion in PC,it may be caused by PNI.The expression of invasion related proteins was up-regulated by NO,wich may change the ability of cell invasion,and the influence of the expression of cell autophagy protein by NO was associated with proliferation and invasion.This study can provide a research theory for the clinical study of nerve infiltration induced by the metabolism pathway of NO in PC. |
