Exploration Of Specific Biomarkers In Tumor Tissue Sample Of The Stage Ⅱ Infiltrating Ductal Breast Cancer

Breast Cancer is one of the common human malignancies in female and hazardous acutely for women health. The incidence of breast cancer is still increasing year by year recently. There was not clear the cause and the mechanism of breast cancer. However, it is important to identify proteins related to the canceration of mammary gland and, to find protein changes during the progression of disease, so as to reveal the mechanism of breast cancer. Consequently, this will in favor of early diagnosis, judging prognosis and making individual treatment of breast cancer.The studies in proteomics are developing rapidly in present years. The sample used is mainly comes from cultured cell line, patients serum, nipple discharge and tissue of breast cancer. The work is easy to cultivate cell line of breast cancer, but it can not completelly reveal the actual condition of the body; And some sdudies have fund that the protein of ental cell having altered with the temporal cultivation of onco-cell in the extraorgan. Secondlly, it was difficullty to detect low abundance protein on account of the effect with redundant albumin and immunoglobulin of serum. The domestic and international reaserch of the proteomics with tissue of breast cancer is still exploring and experiencing because of the interference by the blood,fat and connectivum. Thirdly, it wil increase the difficulty of the experimentation with the material of pathognostic pathological breast cancer. The result will be not known du to the above reason, but it is challenge to extract reflecting total proteins of tissue of pathological changes. Significantly, the sdtudy is able to reveal the real information of the human organism.This experience is also a new way for reaserching proteomics of noumenal tumor.Two dimensional polyacrylamide gel electrophoresis(2-DE) is a primarily protein separated technique in proteome investigation. The quantity of extraction from the protein is being tried to improve by all kinds of ways. The protein extracted from the tissue of breast cancer can represent all the protein of tissue with pathological changes and reflect the case in real intracorporeal situation, but it can be affected by blood, fat and connective tissue etc. The present research object is tissue beside the tumor of breast cancer infitrating ductal carcinoma (the distance to the tumor >3cm, confirmed by pathologic result). It will be optimized on extracting and handling to the samples, quassation of cells and tissue, preparing the solution (principal the associated employment of denaturing agent, surface active agent and reductant), methods of precipitation and staining to acquire SDS-PAGE electropherogram with superior quality and improve reproducibility and stability. The present study will provide foundation for the next research of 2-DE.We have made the comparative proteome analysis with five tissue of the stageâ…¡infiltrating ductal breast cancer and the paired normal glandular tissues adjacent to tumors were carried through. Namely, the total proteins of tissue of the stageâ…¡infiltrating ductal breast cancer and the paired normal glandular tissues adjacent to tumors were separated by means of Immobilized pH gradient (IPG) based 2-DE and silver staining. The differential expression proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spec- trometry (MALDI-TOF-MS). The results were as follwing: The results showed that well-resolved, reproducible 2-DE patterns of tissues of the stageâ…¡infiltrating ductal breast cancer and the paired normal glandular tissues adjacent to tumors were obtained.The spots of 3 differential expression proteins have been fund.Through analysing spots of 3 differential expression proteins by analysis of peptid finger printing patterns, we have identified some relative proteins with gene expression, activity of generation cycle,cell differentiation,tumorous growth,secretory activity of extracellular matrix,tumor metastasis and palindromia etc in breast cancer.It is essential to authenticate the reliability of having identified proteins. The expressive condn of S100 calcium binding protein A11(00A11)nd cllagen alpha-3(VI) chain precursor will be made by technique of immunity class and Western Blot etc to analyse their contribution futurely and deeply for carcinogenesis and development of breast cancer.The results of this study suggest that the proteomics is an effective means in looking for and identifying anomal expressive protein of breast cancer and specific biomarkers in the tumor. The research of differential proteomics with mammary gland tissue of identical clinical stage, isologous patho-type and coimmune is also reproducibility and homogenicity. The specific biomarkers in the tumor are universal representative significance. We analyse and identify the deferential proteins with 2DE and MALDI-TOF-MS and find related protein marker in carcinogenesis, development and metastasis. It will accumulate experience for exploratory of poteomics in solid tumor tissue.