Standardization for Transcriptomic Molecular Markers to Screen Human Colon Cancer

Cancer Genomics - Proteomics

STEFAN P. MARCUARD¹  WADE NAZIRI⁶  DENNIS R. SINAR⁷  FARID E. AHMED⁵  DONALD T. LYSLE³  GORDON FLAKE²  STEPHANIE IJAMES³  PAUL W. VOS⁴ 

[1] arolina Digestive Diseases PA, Greenville, NC, U.S.A.arolina Digestive Diseases PA, Greenville, NC, U.S.A.arolina Digestive Diseases PA, Greenville, NC, U.S.A.;aboratory of experimental Pathology, National Institute of Environmental Health Sciences, Research Triangle Park, NCaboratory of experimental Pathology, National Institute of Environmental Health Sciences, Research Triangle Park, NCaboratory of experimental Pathology, National Institute of Environmental Health Sciences, Research Triangle Park, NC;epartment of Psychology, University of North Carolina, Chapel Hill, NCepartment of Psychology, University of North Carolina, Chapel Hill, NCepartment of Psychology, University of North Carolina, Chapel Hill, NC;epartment of Biostatistics, School of Allied Health Sciences, ECU, Greenville, NCepartment of Biostatistics, School of Allied Health Sciences, ECU, Greenville, NCepartment of Biostatistics, School of Allied Health Sciences, ECU, Greenville, NC;epartment of Radiation Oncology, Leo W. Jenkins Cancer Center, The Brody School of Medicine (BSOM) at East Carolina University (ECU), Greenville, NCepartment of Radiation Oncology, Leo W. Jenkins Cancer Center, The Brody School of Medicine (BSOM) at East Carolina University (ECU), Greenville, NCepartment of Radiation Oncology, Leo W. Jenkins Cancer Center, The Brody School of Medicine (BSOM) at East Carolina University (ECU), Greenville, NC;arolina Physicians PA, Greenville, NCarolina Physicians PA, Greenville, NCarolina Physicians PA, Greenville, NC;epartment of Internal Medicine, The Brody School of Medicine (BSOM) at East Carolina University (ECU), Greenville, NCepartment of Internal Medicine, The Brody School of Medicine (BSOM) at East Carolina University (ECU), Greenville, NCepartment of Internal Medicine, The Brody School of Medicine (BSOM) at East Carolina University (ECU), Greenville, NC

关键词: Adenocarcinoma;    colonocyte;    diagnosis;    inflammatory bowl diseases;    laser capture microdissection;    RT-qPCR;    RNA;    staging;   

DOI  :  

来源: Delinasios GJ CO

【 摘 要 】

Establishing test performance criteria for a transcriptomic colon cancer marker approach must be carried out in a standardized fashion in order tso ensure that the test will perform the same way in any laboratory, anywhere. Condition of sample preservation and shipping prior to total RNA extraction is critical, and we recommend preserving stool samples in an appropriate preservative and shipping them in cold packs so as to keep stools at 4°C. It is not necessary to isolate colonocytes to obtain adequate RNA for testing. It is, however, important to obtain samples from both mucin-rich and non-mucin rich to have a good representation of both left- and right-side colon cancers. Employing a commercial total RNA extraction kit that contains an RLT buffer from Qiagen Corporation (Valencia, CA, USA) removes bacterial RNA from stool preparations and results in a high yield of undegraded RNA of human origin. Genes selected based on the enormous resources of NCI's Cancer Genome Anatomy project give good results. Primers for PCR should span more than one exon. Use of semiquantitative PCR, preferably with several reference housekeeping genes of various copy numbers, depending on tested genes, should enhance confidence in the quantitative results. Having standardized the testing conditions in our ongoing work, it is now imperative that a larger prospective randomized clinical study utilizing stool and tissue samples derived from several control and colon cancer patients, to allow for statistically valid analyses, be conducted in order to determine the true sensitivity and specificity of the transcriptomic screening approach for this cancer whose incidence is on the rise worldwide.

【 授权许可】

Unknown