Study On Different Proteomics Of Breast Cancer

Breast cancer is one of the most frequent female tumors. The incidence has an increasing trend all of the world. Although China belongs the relative low incidence country, the incidence and mortality in the big coastway cities is elevating. The breast cancer incidence in Tianjin has climb from 16.91/100,000 in 1981 to 42.25/100,000 in 2000, the rude incidece increase 150%. By now the incidence ranks the second among all female tumors. And then the age-specific incidence showed the occurring peak of the breast cancer presented a younger trend.The epidemiological study on the breast cancer etiology considered that the occurring was related with some physiology, bearing and genetic factors partly, the others involved with the enviroment risk factors. Recent researches on breast cancer carcinogenesis had deeped companied the molecular biological technique development and updating, some breast cancer susceptible and related genes had founed. However the gene function acter was protein indeed, all of the phenotypes were not interpreted completly from the gene level, so the molecular biological research fields entered into the proteomics phase.There was less than 10 years on proteomics research from the term was produced, the breast cancer related database was not complete, the breast cancer proteomics research belongs fashion and hot project in world. The cell model was a powerful tool in the proteomics research fields because of the standard cell culture, the comparative results and controleble experimental condition. So the aim of this study is to compare the difference in breast cell line and the normal breast epithelial cell line and to search the different proteins to elucidated the proteome change in the precedure of the breast cancer occurring and development.Part I : Separation of proteins-establishing and optimizing the technique of the two-dimension gel electrophoresisObjective1. To establish and optimize the research methods for proteome analysis of separating proteins of cell line and serum by using the two-dimension gel electrophoresis (2DE).2. To search the differentially expressed protein spots on the 2DE pattern from the cell line derived from the breast cancer and the normal breast epithelial cell and serum derived from the breast cancer patient and the health control donors.Methods1. The breast invasive ductal cancer (IDC) cell line T47D and the normal breast epithelial cell line HBL-100 were cultured, the total proteins were extraced after cells was lysated. The samples were analized using 2DE respectively. The first dimension was carried by IPG pH gradient isoelectric focusing system, the second dimension was separatd by vertical SDS-PAGE, the last step was steined by silver or coomas brilliant blue.2. The 2DE patterns were analized and the spots were matched and compared by ImageMaster image analysis software. The statistical method was applized to evaluate the 2DE results.3. All of the experimental procedure were optimized and improved.Results1. The good reproducible results were acquired using the optimized 2DE technique. The protein spots which locate the area of Mr 20~67 kDa and pi 4~8 were separated well.2. The mean resolution were 804on the breast cells 2DE patterns, the match rate was 79.7% between the T47D and HBL-100 cell line, 23 protein spots were differentlly expressed sinificantlly.3. The mean resolution were 257 on the serum 2DE patterns, the match rate reach at 95.6%,27 protein spots were differentlly expressed sinificantlly.Conclusion1. The proteins were separated successfully for proteome research of the breast cancer using the optimized 2DE technique.2. The 2DE patterns were similar between the breast cancer cell line T47D and the normal breast epithelile cell line HBL-100. But there were the differentially expressed proteins between them.3. The 2DE patterns were more similar in human serum between the primary breast invasive ductal cancer patients and the...