The Study Of Cancer Correlative Genes (AP-2β,TP53,NOD2)

In this study, we focus on transcription factor AP-2β, TP53 and nucleotide oligomerisation domain(NOD2), which are correlative with cancer.In part 1 of this study, a DNA-based Surface Plasmon Resonance (SPR)biosensor has been developed for the detection of TP53 mutation using the inexpensive and commercially available instrument, SPREETA^TM SPR-EVM-BT, from Texas Instruments. A direct immobilization procedure, based on the coupling of thiol-derivatised oligonucleotide probes (Probe-C6-SH) to bare gold sensor surfaces, was optimized using synthetic oligonucleotides. Hybridization reactions between the immobilized probe and a short sequence (26 mer) complementary, non-complementary and one point mutation DNA were then investigated. The main analytical parameters of the sensor system were studied in detail including selectivity, sensitivity, reproducibility and analysis time. Finally, the sensor system was successfully applied to polymerase chain reaction (PCR) amplified real samples, DNA extracted from both normal, wild type, (Jurkat) and mutated (Molt 4), carrying the mutation at codon 248 of the TP53 cell lines. The results obtained demonstrate that the DNA-based SPR biosensor was able to distinguish sequences present in the various samples that differ only by one base and hence it appears to be a strong candidate technique for the detection of gene mutation.In part 2 of this study, we were curious to explore whether thereexist any possible association between NOD2/CARD15 3020insc frameshift mutation and colorectal cancer, or not. Colorectal cancer (CRC) is generally thought of as a disease of older persons, with more than 90% of patients being diagnosed after age 55 years. Individuals diagnosed with Crohn's disease (CD) have been shown to have an increased risk of CRC. A susceptibility gene for CD was identified on chromosome 16 and was . initially named NOD2 and later renamed CARD15. The C-insertion at nucleotide 3020 in the leucine-rich repeat region of NOD2 results in a frameshift in 10th LRR followed by a premature stop codon. This truncation mutation may result in the inability to active NF-kB in response to bacterial lipopolysaccharides, suggesting an association between the innate immune response to intestinal bacteria and pathogenesis of CD. Given this biological evidence, it seems there is a possible association between 3020insC in NOD2/CARD15 gene and CRC at an older age. A previous study by Kurzawski et al, reported the presence of the 3020insC mutation appears to increase the risk of developing CRC at an older age. But their results arose up an interesting debate about the relationship between the 3020insC NOD2 polymorphism and colorectal cancer in a recent issue of Cancer Research. In this study, we tested the 3020insC NOD2 polymorphism of an unselected consecutive series of colorectal cancer patients at an older age (in the range of 50 to 93 years old) and healthy controls. Our experiment facts...