Expressions And Signification Of CFTR And SPINK1 In Chronic Pancreatitis And Pancreatic Cancer

Chronic pancreatitis is a disease which is characterized as pancreatic inflammation and progressive fibrosis. The cause of it are extremely complex, and there are many factors closely related to ,such as alcohol abuse,biliary disease, smoking, endocrine,metabolic,protein and fat diet,diabetes and so on. In our country, the primary factor is biliary tract disease ,while in Europe and Unites countries,the major factor is alcohol abuse. Now more and more scholars study the pathogenesis of the genetic susceptibility in chronic pancreatitis. In 1996, Whitcomb found that hereditary CP was due to the mutation of Cation of the chymotrypsin original is hydrolysis food which contained Lai-arginine residues of proteins, It also play a key role in the process of activate/inactivate other enzmes. As the different experimental condition about the pathogenesis of genetic factors in chornic pancreatitis,also requires further study.Pancreatic cancer is mainly malignant pancreatic exocrine ,which is one of highly malignant digestive tumors. As the complex causes and lack the specific diagnostic criteria, early diagnosis of pancreatic cancer is very difficult. Once obvious clinical symptoms have appeared , it will be at a late stage and miss the best treatment opportunity .The etiology and pathogenesis of pancreatic cancer is currently unknown, and Clinical data analysis it is a result of varieties of long-term factors.The Factors associated with pancreatic cancer contains smoking, alcohol consumption, diabetes, chronic pancreatitis and family history of cancer . With the development of modern molecular biology techniques, the incidence of pancreatic cancer, metastasis and invasion mechanism has entered the genomic level. CFTR and SPINK1 is the two newly identified proteins associated with chronic pancreatitis and pancreatic cancer , CFTR protein is a Cl- channel protein, mediated Cl—HCO3-transport, the function and expression of CFTR protein ensure a normal PH value of pancreatic juice,and have the role of diluting and alkaline pancreatic juice. And then ensure pancreatic juice plays a normal physiological function, SPINK1 protein is trypsin inhibitor, abnormal expression of SPINK1 protein causes the abnormal activation of trypsin. As we all know, chronic pancreatitis associated with abnormal activation of trypsin. Thus, SPINK1 is seen as the first internal protection line of pancreatic tissueThis study is to discuss the expression and signifcance of CFTRand SPINK1 protein in patients with chronic pancreatitis and pancreatic carcinoma, provide experimental data for the prevention and early diagnosis and treatment of chronic pancreatitis and pancreatic carcinoma.In this study,immunohistochemistry (SP method) was used to detect the expression of chronic pancreatitis and pancreatic carcinoma.and then carried out fiow cytometry semi-quantitation detected the protein expression,further confirmed that protein expression.We collect paraffin of normal pancreas, chronic pancreatitis and pancreatic cancer tissues for 10 cases, 20 cases, 30 cases in First Hospital of Jilin University and Department of Pathology and Anatomy School of Basic Medical Pathology. They were cuted into 4um thick slices, leaving one for HE staining,others for immunohistochemical staining,according to SP method steps. Microscope, brown particles in cells is positive expression .Statistical results forχ2 test, P <0.05 was considered statistically significant. CFTR protein was mainly localized in the cytoplasm of pancreatic ductal epithelial cells, the expression of the cell membrane is small amount .In normal pancreatic tissue ,it was positive expression, the expression in chronic pancreatitis and pancreatic cancer was significantly weaker than in normal pancreatic tissue(p<0.05). SPINK1 protein was mainly localized in the cytoplasm of pancreatic acinar cells. In normal pancreatic tissue it was positive expression, chronic pancreatitis and pancreatic cancer were significantly weaker than normal pancreatic tissue, there was significant difference (P <0.05). Using Spearman correlation analysis to analyze the expression of CFTR and SPINK1-related trends, the result was a positive correlation between the expression.Collected six cases of normal pancreatic tissue and 20 cases of pancreatic carcinoma, flow cytometry was used for protein semi-quantitative, Each specimen cut into 50um×6 sheets, according to a single cell suspension in paraffin-embedded tissue preparation steps: dewaxing, hydration, digestion, filtration, centrifugation, washing, made into single cell suspension With first antibody (CFTR and SPINK1) were incubated for 30 minutes at room temperature, PBS wash and then centrifuged, With FITC labeled secondary antibody from light for 30 minutes at 4℃Washing, centrifugation and then resuspended in 150ul PBS for flow cytometry. According to the fluorescence index for protein quantitative According to the experimental group FI value greater and smaller than the upper limit of fluorescence intensity in normal pancreatic cells, were judged to be positive and negative expression. The experimental results show, CFTR and SPINK1 protein show positive expression in the normal pancreatic tissue. Decreased expression in pancreatic cancer, compared with normal pancreatic tissue were significantly different. (p<0.05) show that CFTR and SPINK1 protein correlated with pancreatic cancer. This result further confirmed by flow cytometry.CFTR protein expression in normal can ensure that the dilution and alkalinization of pancreatic juice, When the CFTR protein expression is abnormal, pancreatic juice will concentration and acidification the change of PH value can lead to inflammation of pancreatitis and abnormal cell proliferation and differentiation.The abnormal expression of SPINK1 protein relieved the inhibition of trypsin, so that abnormal activation of trypsin, leading to the occurrence of pancreatitis inflammation. SPINK1 protein abnormal expression in pancreatic cancer, maybe directly involved in the development of pancreatic cancer or as a modification factor reduces the threshold of pancreatic cancer, the specific pathogenic mechanism needs further study.The experimental study provided experimental basis for exploring the complex causes of pancreatic diseases in China, developed new ideas for chronic pancreatitis and pancreatic cancer in the early diagnosis, clinical prevention and treatment,carried out useful theoretical exploration for the gene therapy of pancreatic disease.