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Genetic Instability in Patients with Pancreatic Cancer Analyzed by SCARs and Electrochemical Sensors by Chaoyang Wu, Qiang Yi, Fengqing Wang, Qicai Liu, Liqing Lin, Ailin Liu, Qingquan Chen

Background: Pancreatic cancer is an aggressive disease and the fourth most common cause of cancer death across the globe. It is often not diagnosed until it is advanced. It is necessary to establish a new technology to detect DNA instabilities during the progression of pancreatic cancer and to screen for new molecular markers coupled to putative unknown oncogenes.
Methods: A total of 25 pancreatic cancer tissue specimens were analyzed by sequence-characterized amplified regions (SCARs), including two pathological types (pancreatic ductal adenocarcinoma and neuroendocrine carcinoma). There were 41 random primers and eight long fragment primers used for PCR amplification, and the difference of dNTPs consumptions were detected by nano-electrochemical sensors. Once both dATP and dGTP are significantly different in oxidation current (reduce or increase simultaneously), separate the different genes by electrophoresis, then clone and sequence the genes, and carry out homology analysis.
Results: Both dGTP and dATP showed good oxidation behavior on the carbon nanotube modified glassy carbon electrode. There were 32 different fragments in malignant tissues compared with normal control, among them a SNP located in 5'UTR of the leucine zipper protein 4 gene which is significantly correlated with pancreatic cancer (OR = 9.50) and it was confirmed by direct sequencing.
Conclusions: SCARs combined with the nanoelectrochemical sensor can be used for screening genetic instabilities in pancreatic cancer, and leucine zipper protein 4 was a novel pancreatic cancer-related gene.

DOI: 10.7754/Clin.Lab.2013.130225