Establishment Of Two-dimension Gel Electrophoresis Protein Profiles And Differential Aanalysis From Human Pituitary Adenoma With And Without Invasivness

PURPOSE:The investigation is the establishment of credible two-dimensional electrophoresis maps from human pituitary adenoma with and without invasiveness for the first time,the identification of differential expression proteins by means of proteomics methods,and to provide a base for identifying proteins related to human pituitary adenoma with invasiveness.MATERIALS AND METHODS:20 samples were collected after surgery from patients after postoperative patho-evaluation(Xiangya Hospital,Central South University).The invasive group 10 patients,and the noninvasive group 10 patients.The total proteins of the human pituitary adenomas with and without invasion were separated and visualized by immobilized pH gradient(IPG)-based two-dimensional polyacrylamide gel electrophoresis(2-DE)and Coomassie brilliant blue staining.2-D gel image analysis was performed with Image Master 2-DE analysis software.The selected differential protein spots were identifed by peptide mass fingerprint(PMF)based on matrix- assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS) and database searching.RESULTS:①Credible 2-DE protein profiles from human pituitary adenoma with and without invasiveness were obtained.2-DE for a selected sample randomly from each of the 2 groups was repeated 3 times,respectively,for the reproducity of the method.The image analysis showed:For the invasive group,the average proteins spots of 3 gel were 1080±24 spots,three were 909±45 spots matched,the average match rate was 84.3%.For the noninvasive group,the average proteins spots of 3 gel were 1035±28 spots,three were 828±56 spots matched,the average match rate was 79.5%.The average position deviation of matched spots was 1.563±0.259mm in IEF direction,and 1.088±0.206mm in SDS-PAGE direction.②For the invasive and noninvasive group,the average proteins spots of two gel were 899±58 spots matched,the average match rate was 83.3%.Thirty differential protein spots(30\99) were identified firstly by peptide mass fingerpriting,which invovle in the regulation of cell cycle and signal transduction.CONCLUSIONS:The credible 2-DE protein profiles from human pituitary adenoma with and without invasiveness were established for the first time,and identifide sonme proteins,such as oncogene DJ-1,HSP27 and so on,which related with invsiveness of human pituitary adenoma PURPOSE:The purpose of this study was to research prospectively whether oncogene DJ-1 and HSP27 were associated with invasiveness of the human pituitary adenoma,and discuss the possibility of taking them as the markers to indicate invasiveness and prognosis.METHODS:20 samples were collected after evaluation.All tumors surgery from were graded patients after full endocrine and staged according to Hardy-Wilson and Knosp classification,using magnetic resonance scanning.Western blot staining was adapted to semi-quantify the expression of DJ-land HSP27.SPSS 13.0 for windows was employed to analysis the correlation of DJ-land HSP27 with the invasiveness of pituitary adenoma.RESULTS:1.The positive rate of DJ-1 and Hsp27 were 70%(7/10) and 80%(8/10)in 10 invasive pituitary adenoma,respectively.The invasive group had a significant higher expression of DJ-1 and HSP27 than noninvasive group 10%(1/10),10%)(1/10);2 There was a positive correlation between the expression of DJ-1,HSP27 and the invasiveness of pituitary adenoma from Spearman rank correlation test(P<0.05).CONCLUSION:The invasive group had a significant higher expression of DJ-1,HSP27(70%,80%)than noninvasive group(10%,10%)(P<0.05).The proliferative activity and abnormal expression of oncogene DJ-1 and HSP27,which play a comparatively significant role in tumorigenesis and progression of pituitary adenoma.Combineded detection of these proteins have important effects on invasiveness determination,prognosis and treatment.The positive correlation between expression of DJ-1,HSP27 and the invasiveness of pituitary adenoma was significant.The expression of HSP70 may be taken as one of the markers to indicate invasiveness and prognosis.These data may be used to be the biomarkers and to further study on invasiveness of the human pituitary adenoma.