A Proteomic Study On Differences Among APC Mutation In Colorectal Carcinoma Tissues

Objective: In order to find out the proteomic differences between APC mutation carcinoma tissues and no APC mutation carcinoma tissues, and between the carcinoma tissues and normal mucous epithelium. We use two-demensional gel electrophoresis and mass spectrometry in APC mutation carcinoma tissues, no APC mutation carcinoma tissues and normal mucous epithelium. Hope fully, our study would be useful in illuminating the process of cancerization of APC mutation colorectal carcinoma.Methods: The fresh sporadic carcinoma tissue and mormal mucous epithelium tissue was cut off after the specimen was resected from the body immediately. APC gene was amplified by using polymerase chain reaction single strand conformation polymorphisms ( PCR-SSCP). Then, the tissue were classified into three groups: APC mutation carcinoma, no APC mutation carcinoma and normal mucous epithelium tissue. Protein separating was performed using two dimension gel electrophoresis with Amershan 18cm linear PH 3-10 immobilized PH gradient (IPG) strips and SDS-PAGE. After silver-staining and preserving, the images of the gel was analyzed by using PD-Quest 7.3 software. The expressed protein spots differentially in each group was incised from the gels. After degested by trypsin, each peptide mass finger printing (PMF) was acquired by matrix assisted laser desorption / ionization time of flight mass spectrometry (MALDI-TOF-MS) above this. The corresponding proteins were identified by data searching in the Mascot-database.Results: 14 (45.2%) cases coloretalcarcinoma with APC mutation was find out in all 31 cases. By PCR-SSCP, 2-DE and MALDI-TOF-MS, most of the gels that we acquired was clearly. Each gels contained approximate 590-980 spots. The protein spots concentrated in the range of 30-100kDa and PH 3.5-7. The average number of the protein spots that we acquired was 814 + 56 in APC...