BRCA 1 AND BRCA 2 Gene Mutations and Increased Risk Of Breast Cancer Abstract References Research Proposal Obed K. Agyei and Natalie A. Rich Department of Biology, North Carolina State University Conclusion Barnes R, Masood S, Barker E, Rosengard AM, Coggin DL, Crowell T, King CR, Porter-Jordan K, Wargotz ES, Liotta LA. Low nm23 protein expression in infiltrating ductal breast carcinomas correlates with reduced patient survival. Am J Pathol 1991 Aug;139(2): Hofmann, W., and P M. Schlag. "BRCA1 and BRCA2 - Breast Cancer Susceptibility Genes." J Cancer Res Clin Oncol (2000): Web. 4 Mar "Cancer Risks in BRCA2 Mutation Carriers." Journal of National Cancer Institute (1999): Web. 4 Mar Image courtesy of prweb.com Center for Disease Control and Prevention. “Breast Cancer Statistics.” (accessed March, 2011). Kretschmer C, Sterner-Kock A, Siedentopf F, Schoenegg W, Schlag PM, Kemmner W. Identification of early molecular markers for breast cancer. Mol Cancer 2011 Feb 11;10(1):15. Smith, Margaret., Susan Fawcett, Emanouil Sigalas, Richard Bell, Sophie Devery, Nikolina Andrieska, and Ingrid Winship. "Familial Breast Cancer: Double Heterozygosity for BRCA1 and BRCA2 Mutations with Differing Phenotypes." Familial Cancer (2008): Web. 3 Mar Veltman, J., R Mann, and T Kok. "Breast Tumor Characteristics of BRCA1 and BRCA2 Gene Mutation Carriers on MRI." Eur Radiol (2008): Web. 4 Mar American Cancer Society. “Breast Cancer Overview.” (accessed March 5, 2011). Hall, Michael J., Julia E. Reid, and Lynn A. Burbidge. "BRCA1 and BRCA2 Mutations in Women of Different Ethnicities Undergoing Testing for Hereditary Breast-ovarian Cancer." Cancer (2009): Web. 4 Mar Breast Cancer affects nearly 273,000 people every year in the United States, and women account for almost 95% of the individuals affected. Private companies and the U.S. government spend almost $1 billion on breast cancer research annually to improve its diagnosis and treatment. Breast cancer is a complex, deadly disease highly associated with genetic abnormalities. Since there are numerous types and sub-types of breast cancer, it is very difficult to understand the basic cause of the disease. The objective of this paper is to review how genetic abnormalities correlate with different types of breast cancer. Research in this field has led scientists to determine the main types of breast cancer that manifest in humans and the genetic causes that lead to these manifestations. Much of the research in this field involves experimentation with genes. BRCA1/2 genes were examined among patients with breast cancer and those without the disease. DNA analysis of BRCA1/2 genes was performed. Majority of the patients had mutations on the BRCA1/2 genes while the other control group showed no sign of genetic mutations. Mutations of the BRCA1/2 genes are typically hereditary, but other factors such as radiation can trigger mutations. Types of breast cancer are associated with the part of the breast whose cells are being transformed due to mutated genes, either being the ducts, lobules etc. Further elucidation of the affects of mutated BRCA1/2 is essential for understanding human breast cancer manifestations and future treatment of this complex disease. As seen in Figure 1 and 2, breast cancer incidence rate is higher in Caucasians women but however, more African-American women tend to die from breast cancer than any other ethnic group. This disturbing findings cautioned us to design an experiment that was geared toward examining the cause of this high mortality rate. Goal 1: to determine the rate of BRCA1 and BRCA 2 gene mutations based on a patient’s ethnicity and age. Goal 2: Use Needle Breast Tissue Biopsy to extract cancerous tissues from a total of breast cancer patients. Cells from six healthy patients will also be collected. Goal 3: Use siRNA gene slicing technology to temporary knockdown each of the BRCA genes from the T47D cell lines to determine its effect on mutation rates. Goal 4: Consult a geneticist to assist in the permanent removal of both the BRCA 1 and BRCA 2 genes from the breast cancer cell lines. We expect to find a decrease in mutation rate when BRCA genes are permanently deactivated. A deactivated BRCA gene will prevent all necessary mutations in the TD47 cancer cell lines. We expect to find significant differences in BRCA 1 and BRCA 2 mutation rate among African American and Caucasian women. General Goal Statement Mutations of the BRCA 1&2 tumor suppressor genes have a tremendous effect of the type of breast cancer that an individual can develop, depending on the cell tissues where the BRCA 1&2 genes are being mutated. Studies have shown that mutations in certain genes can lead a person to be more susceptible to developing breast cancer. The most commonly studied genes are the BRCA1 and BRCA2 genes. BRCA 1 and BRCA 2 are human tumor suppressor genes that produces proteins to prevent uncontrollable cell growth. In the absence of BRCA genes, the human body risk the chance of preventing mass cell division. The purpose of this study is to review the risk of BRCA mutations, in comparison to other factors such as protein levels, diet, health, and weight that may contribute to the cause or prevention of breast cancer and the type of breast cancer associated. Introduction Materials and Methods Figure 1 represent the survival rate of patients with low and high nm-23 proteins.(Barnes et al 1991). Figure 2 illustrates breast cancer risk in women. Gray column show risk in women with a strong family history of breast cancer estimated by Easton et.al. (1995); Gray stripped column shows risk in women carrying BRCA 1 or BRCA 2 gene mutation estimated by Struewing et al.(1997); White column shows risk in general population. (Hofmann et al 2000). *Black *White *(Center for Disease Control and Prevention, 2011). Rates per 100,000 persons among 19 age groups. Figure 4. Shows female breast cancer mortality rates in the U.S with respect to ethnicity. (CDC) Results Infiltrating ductal carcinomas with low nm-23 protein had a low rate of survival. Human Ductal Carcinoma In-Situ genes express markers similar to invasive ductal carcinomas in mice. Invasive ductal carcinomas and medullary carcinomas are positive for Cytokeratin 5 and 6 proteins Infiltrating Lobular Carcinoma is positive for 92.7% of estrogen and has a higher presence of p53 tumor suppressor gene Examination of Inflammatory breast cancer tissues showed that the genes had recurrent amplifications than the genes in non-inflammatory breast cancer tissues BRCA1/2 mutations are commonly found in breast cancer patients. Mutations in the BRCA1/2 genes are found in people with a family history of breast cancer. BRCA1/2 gene mutations increase the rate of tumor growth in patients with breast cancer. BRCA1/2 genes increase the risk of a person developing breast cancer by 85%. Majority of BRCA ½ gene mutations are found in ancestral groups The amount of protein levels and gene mutations can determine a patient’s rick of developing breast cancer and chances of survival. As it can be inferred from Figure 1, low levels of nm-23 (a specific type of protein) can reduce the survival rate of infiltrating ductal carcinoma. These findings by Barnes were supported by three pathologists who had similar conclusions. They concluded that high levels of nm-23 protein will increase a patient’s survival of infiltrating ductal carcinoma. Since infiltrating ductal carcinoma is the most common type of all breast cancer, this research is very significant because a progress can be made to reduce its popularity. Few steps that could be taken to reduce the popularity of infiltrating ductal carcinoma include developing medicines that have high concentrations of nm-23 protein. In comparison to BRCA genes, similar conclusions could be drawn because a mutation in BRCA 1 or BRCA 2 gene, as seen in Figure 2, will reduce the production of protein by the BRCA genes. A reduction in protein production cannot prevent cells from stopping its mass division which ultimately ends up in the build up of tumors. In conclusion, BRCA 1 / 2 genes are highly associated with an individual’s risk of developing breast cancer. Mutation of these genes reduces cell signaling during cell division, thus increasing uncontrollable cell division in the body. Acknowledgements This work was supported by the College of Agriculture and Life Sciences, Department of Biology at North Carolina State University, Dr. Miriam Ferzli and Elizabeth Overman. Gene Expression Profiling: [To accurately determine the state of cancerous tumors.] A set of human DCIS and invasive mammary tumors were analyzed using gene expression profiling. Gene Expression of TP53 genes were examined in normal breast epithelial cells and mutated breast epithelial cells. A test was conducted on somatic cells affected by cancerous genes to identify their phenotypes. DNA analysis of the BRCA1 and BRCA2 genes was performed over the course of ten years on women from various ethnic groups. A genetic test was conducted in mice with one or more mutated genes. DNA and RNA were extracted from frozen samples of invasive carcinoma tissues and analyzed for genomic expressions. Population Study Families with the BRCA1 and BRCA2 mutations were investigated to determine the risks of developing breast cancer A database of patients with early stages of breast cancer were studied to determine the status of the TP53 tumor suppressor gene. A nationwide study was conducted to help determine non-genetic factors for developing breast cancer in premenopausal and postmenopausal women. Protein Level Analysis Infiltrating ductal carcinoma were collected from the University of Jacksonville to be accessed for protein staining patterns. Medullary carcinomas of the breast were obtained and examined for the presence of cytokeratin Figure 3. A representation of female breast cancer incidence rates in the U.S with respect to ethnicity. (CDC) High level of nm23 Low level of nm23