Capillary electrophoresis in analysis of DNA variations in rectal cancer.
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AbstractFor colorectal cancer surgery is the main and most efficient treatment. In spite of this many patients later suffer from metastases or tumour recurrence. One cause may be peritoneal dissemination of free cancer cells exfoliated from the serosa surfaces of primary cancers.
We have developed and validated a denaturing capillary electrophoresis (DCE) method that is very sensitive in detecting mutation. The DCE method is simple, with only a two-step protocol PCR and electrophoresis. The method is now high throughput with the convertion to commercially automated single and multicapillary instruments.
We used this method to detect mutations in the k-ras gene with DNA from cells obtained by lavage. Patients with k-ras mutated cells in the lavage immediately after surgery have a reduced life expectation. Detection of exfoliated cells in the abdominal cavity may be a useful diagnostic tool to improve the staging and eventually characterize patients whom could benefit from aggressive multimodal treatment of rectal cancer. We have also applied the DCE method to detect selected DNA variations important in the adenoma-carcinoma sequence and in pharmacogenomic aspects of rectal cancer patients.
Kristensen AT, Bjørheim J, Minarik M, Giercksky K-E, Ekstrøm PO: Detection of mutations in exon 8 of TP53 by temperature gradient 96-capillary array electrophoresis.Biotechniques 2002, 33: 650-653. Abstract.
Bjørheim J, Abrahamsen TW, Kristensen AT, Gaudernack G, Ekstrøm PO: Approach to analysis of single nucleotide polymorphisms by automated constant denaturant capillary electrophoresis.Mutat Res 2003, 526: 75-83. Abstract.
Kristensen AT, Bjørheim J, Wiig J, Giercksky KE, Ekstrøm PO: DNA variants in the ATM gene are not associated with sporadic rectal cancer in a Norwegian population-based study. Int J Colorectal Dis. 2004 Jan;19(1):49-54.. Abstract.
Kristensen A.T, Wiig JN, Larsen SG, Giercksky KE, Ekstrøm PO. Association study of DNA variations in genes of the folate metabolism with rectal cancer in a Norwegian population. Submitted January 2007.
Kristensen A.T, Wiig JN, Larsen SG, Giercksky KE, Ekstrøm PO. Molecular detection (k-ras) of exfoliated tumour cells in the pelvis is a prognostic factor after resection of rectal cancer? Submitted February 2007.