Identification Of Potential Biomarkers In Breast Cancer Tissues Among Malaysian Patients
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Date
2011-07
Authors
Lim, Chu Ai
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Journal ISSN
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Publisher
Universiti Sains Malaysia
Abstract
Breast cancer is the third most common cancer worldwide and is the most common cancer among Malaysian women. In year 2006, there were 3525 new cases of female breast cancer registered with Malaysia National Cancer Registry. Protein is the expression component of cells that regulates cellular activity. By comparing healthy and cancerous tissues at the protein level, it is possible to identify the potential protein targets that can serve to detect, treat, monitor, and for prognosis of breast cancer. In this study, proteins were extracted from 25 pairs of breast normal and cancerous tissues from patients who suffered from infiltrating ductal carcinoma (IDC). These proteins were separated according to their pI‟s and molecular weights. The protein spots were analyzed and the protein spots of interest were excised from the gel. After that, they were digested with trypsin to produce fragment of peptides that can be analyzed using LC/MS/MS. The data obtained from LC/MS/MS analysis were subjected to protein database search engine for protein identity determination. Twenty-six protein spots from Tris extract and 18 protein spots from TLB extract were identified due to their differential expression between cancerous and normal tissues. Fibrinogen beta chain was the down-regulated protein (>70%) in Tris extract, while 80K protein H precursor, tubulin, beta polypeptide, calreticulin variant and protein disulfide-isomerase (PDI) were the up-regulated proteins (>70%) in TLB extract. PDI and heat shock protein 60 (HSP60) were selected for Western blot and immunocytostaining studies to validate the presence of these proteins and their extracellular location. The results showed that PDI was located on the external membrane surface of MCF-7, T47D and MDA-MB-231 cell lines. Meanwhile HSP60 was located on the external membrane surface of MCF-7 and T47D cell lines.
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Keywords
Potential biomarkers in breast cancer tissues , among malaysian patients