Expression And Significance Of Integrin Linked Kinase (ILK) In Pancreatic Adenocarcinoma

IntroductionTumor is a pathological progression consisting of many genes,steps and stages. Pancreatic adenocarcinoma is one of the frequent cancer of the digestive system.Pancreatic adenocarcinoma is the fatal cancer of the digestive system with the worst prognosis.the difficulty of early diagnosis due to its anatomical location,and lack of specific early syndromes,the high potential to infiltrate to the surrounding tissues and metastasize even in the early stage,the poor responsiveness to conventional treatments such as chemotherapy,radiotherapy,Surgery represents the only opportunity for possible cure,but it is restricted to early stage pancreatic adenocarcinoma and most patients who undergo tumor resection show recurrence or distant metastases and die within a few years.the mechanism of invasion and mestastatic of pancreatic adenocarcinoma has becomed a hot topic.As a serine/threonine protein kinase,ILK is a upstream cross point of some tumor-associated factors.ILK has effect on formation,proliferation,invasion and metastasis of tumors by cell adhesion,signal transmit and cell cycle.ILK has became a hot spot in cancer research.The expression of ILK in normal tissues is very limited.but in tumor tissues is noticeably higher than that in the normal tissues,for example in human neurogenic,gastric cancer,prostatic carcinoma,tumor of the ovary,melanoma and lung cancer.it keeps correlation to pathology factors of tumors.Now,there have not reports about the relativity of ILK and pancreatic adenocarcinoma in home and abroad.so we investigated the expression of ILK in pancreatic adenocarcinoma and normal tissue by immunityhistochemistry and wester blot,combined the intact clinial date.then analyzed the relation of ILK and clinical pathology factors.Materials and Methods1.Materials60 pancreatic adenocarcinomas and 32 normal tissue samples were obtained from patients who had surgery in the First Clinical Hospital in China Medical University between 1990 and 2008,fix in 10%paraformaldehyde,embedded in paraffin.10 freshly pancreatic adenocarcinomas and normal tissue samples were obtained from patients who had surgery in the First Clinical Hospital in China Medical University between 2007 and 2008.2.Methods(1)Immunohistochemistry:Detect the ILK expression byimmunohisto-chemistry S-P method.Reasult determination:ILK was a plasm staining protein.under light microscope,we select three high power fields in each chip,and count 100 neoplastic cells in each high power field.three observers independently evaluated the immunostaining results.Cytolymph and cellular membrane were stained flavedo or pallide-flavens is positive。Review the stain ratio of the cell under micro。the stain ratio exceed 30 percent was defined as positive.(2)Western blot:Detect the expression of ILK in pancreatic cancer tissues and noncanceroustissues.Rapidly homogenize tissues(1-2g) in 5 volumes of lysis buffer centrifuge the homogenate(11000rpm,4℃)for 20 minutes to pellet insoluble material. Electrophoresis,transfer printing,then incubated with polyclonal anti-ILK antibody, overnight at 4℃.After incubated with Horseradish peroxidase-conjugated second antibodies.Reasult determination:appearance amethyst vittae is positive,we detect gray scale value with FluorChemV2.0 Stand Alone.3.Statistical analysisAll the data were analyzed with SPSS for Windows 10.0 software.x~2 test was to compare the clinicopathological characteristics with the expressions of ILK.t test was to compare the gray value of Western blot results between the pancreatic carcinomas group and the normal group。Statistical significance was indicated by P＜0.05.Results1.Expression of ILK in pancreatic carcinomas and normal tissues ILK was not detected in normal tissues,but it showed a strong expression in many pancreatic carcinomas tissues.Western Blot Method showed the level of ILK in the tumor tissues is noticeably higher than that in the normal tissues2.the relationship of ILK and clinic pathology factorsThe positive rate of ILK in pancreatic carcinomas tissues was 65%(39/60),The expression of ILK was related to lymphatic metastasis and clinical stage(P＜0.05),but was not related to the differentiation(P＞0.05).Conclusion1.The level of ILK in the tumor tissues is noticeably higher than that in the normal tissues2.In pancreatic carcinoma,the expression of ILK was positive correlation to clinical stage and lymph metastasis and noncorrelation to differentiation.