Abstract
This study investigated the use of DNA amplification fingerprinting (DAF) to identify biomarkers useful in the elucidating genetic factors that lead to carcinogenesis. The DNA amplification fingerprinting (DAF) technique was used to generate fingerprint profiles of a normal human mammary epithelial cell line (MCF-10A) and a human breast cancer cell line (MCF-7). When compared with one another, a polymorphic biomarker gene (262 base pairs (bps)) was identified in MCF-10A but was not present in MCF-7. This gene was cloned from the genomic DNA of the MCF-10A cell line, and subjected to Genbank database analysis. The analysis of the nucleotide sequence polymorphic marker (Genbank account: AC079630) shows that this biomarker has 100% homology with the nucleotide sequence of human chromosome 12 BAC RP11-476D10 (bps 19612-19353). The nucleotide sequence was used for possible protein translation product and the result obtained indicated that the gene codes for hypothetical protein XF2620. In order to evaluate the effects that the 262 bps biomarker would have on the morphology of MCF-7 cells, it was transfected into MCF-7 cells. There were observable changes in the morphology of the transfected cells. These changes included an increase in cell elongation and a decrease in cell aggregation.
Highlights
DNA “fingerprinting” has been used for genome linkage, genetic variation, population and pedigree analysis, forensic identification, localization of disease loci, and epidemiology [1,2,3]
When compared with one another, a polymorphic biomarker gene (262 bps) and others were identified in MCF-10A but were not present in MCF7 (Figure 1)
There were other biomarker genes present, this study focuses only on the 262 bps biomarker
Summary
DNA “fingerprinting” has been used for genome linkage, genetic variation, population and pedigree analysis, forensic identification, localization of disease loci, and epidemiology [1,2,3]. The most common form of cancer among women is breast cancer [10]. It is the second leading cause of mortality among females, the pathogenesis of the disease remains unclear [11, 12]. Most mutations in human malignancies were identified by conventional methods such as single-strand conformational polymorphism (SSCP) and DNA sequencing [13]. Other methods, such as denaturing gradient gel electrophoresis, heteroduplex analysis, and cleavage methods [14] have been used. All these methods are relatively time-consuming, labor intensive, and sequential processes
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