Breast Cancer Treatment biochemical studies aimed at determinin

Biochemical Approaches in Breast Cancer Treatment

Breast cancer is the most common cancer found in women and in Western societies it is the leading cause of early death in women (Cao et al 2004). Most human breast cancers require estrogen for their continued growth, and these cancers have been shown to respond to antiestrogen therapy alone or when it is used in combination with other forms of chemotherapy (Khalkhali-Ellis et al, 2004). Because a large number of women who may be at risk for breast cancer receive hormone therapy, there is a pressing need to discover the role played by estrogen in promoting breast cancer. The tumor suppressor gene maspin is known to be present in high concentrations in normal mammary epithelial cells, which also require estrogen for continued growth. However, maspin is down-regulated in primary breast cancer cells, and is entirely absent in aggressive mammary

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d. Western Blot Analysis: Cytoplasmic proteins were lysed in radioimmunoprecipitation buffer and separated on 7.5% SDS-PAGE then transferred to a polyvinylidene difluoride membrane (Cao et al, 2004). The membrane was incubated with anti-CLA-1 antibody or anti-cyclophilin-A antibody and the binding was visualized by chemiluminescence detection. Proliferation Assay: 104 MCF-7 cells were cultured with 200 ?l DMEM with 10% lipoprotein deficiency serum. The medium was replaced with DMEM plus lipoprotein deficiency serum with or without 100-500 ?g/ml HDL, the cells incubated for 24 h at 37C, the cells were pulsed with 1 ?Ci/well [3H]thymidine (specific activity = 6.7 Ci/mmol), buretted onto glass fiber filters using an automated cell harvester, and counted in a Packard liquid scintillation counter (Cao et al, 2004). Transfection of MCF-7 Cells and Luciferase Reporter Gene Assay: The AP-1 reporter gene and purified reporter plasmid were transected into MCF-7 cells at 60% confluence using a conventional cationic lappaceum transfection method (Cao et al, 2004). All assays were corrected for ?-galactosidase activity and the total protein in each reaction was identical. Twenty ?l aliquots were used for the luciferase assay which was
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Approximate Word count = 4363
Approximate Pages = 17 (250 words per page)