DNA integrity as a potential marker for stool-based detection of colorectal cancer.

BACKGROUND Molecular genetic analysis of DNA in patient stools has been proposed for screening of colorectal cancer (CRC). Because nonapoptotic cells shed from tumors may contain DNA that is less degraded than DNA fragments from healthy colonic mucosa, our aim was to show that DNA fragments isolated from stools of patients with CRC had higher integrity than DNA isolated from stools of patients with healthy colonic mucosa. METHODS We purified DNA from the stools of a colonoscopy-negative control group and patients with CRC and examined the relationship between long DNA fragments and clinical status by determining stool DNA integrity, using oligonucleotide-based hybrid captures with specific target sequences in increasingly long PCR reactions (200 bp, 400 bp, 800 bp, 1.3 kb, 1.8 kb, 24 kb). DNA fragments obtained from CRC patients were compared with fragments obtained from colonoscopy-negative individuals for length and/or integrity. RESULTS DNA fragments isolated from CRC patients were of higher molecular weight (>18 bands detected of a total of 24 possible bands) than fragments isolated from fecal DNA of the colonoscopy-negative control group. CONCLUSIONS The presence of long DNA fragments in stool is associated with CRC and may be related to disease-associated differences in the regulation of proliferation and apoptosis. An assay of fecal DNA integrity may be a useful biomarker for the detection of CRC.

[1]  D. Lieberman,et al.  One-time screening for colorectal cancer with combined fecal occult-blood testing and examination of the distal colon. , 2001, The New England journal of medicine.

[2]  K. Kinzler,et al.  Identification of ras oncogene mutations in the stool of patients with curable colorectal tumors. , 1992, Science.

[3]  P. Pasricha,et al.  Inhibition of apoptosis during development of colorectal cancer. , 1995, Cancer research.

[4]  J. Kumi‐Diaka,et al.  The mediating role of caspase‐3 protease in the intracellular mechanism of genistein‐induced apoptosis in human prostatic carcinoma cell lines, DU145 and LNCaP , 2000, Biology of the cell.

[5]  E. Mohr.  [Fecal occult blood testing for colorectal cancer ]. , 2003, Revue medicale de Liege.

[6]  S Jothy,et al.  Apoptosis, anoikis and their relevance to the pathobiology of colon cancer , 2000, Pathology international.

[7]  T. Byers,et al.  American Cancer Society guidelines for screening and surveillance for early detection of colorectal polyps and cancer: Update 1997 , 1997 .

[8]  N. Kaibara,et al.  Apoptosis as a Prognostic Factor in Colorectal Carcinoma , 1998, Surgery Today.

[9]  K. Kinzler,et al.  Apoptosis and APC in colorectal tumorigenesis. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[10]  C. Cordon-Cardo,et al.  Genetic and Molecular Markers of Urothelial Premalignancy and Malignancy , 2000, Scandinavian journal of urology and nephrology. Supplementum.

[11]  T. Byers,et al.  American Cancer Society guidelines for screening and surveillance for early detection of colorectal polyps and cancer: update 1997. American Cancer Society Detection and Treatment Advisory Group on Colorectal Cancer. , 1997, CA: a cancer journal for clinicians.

[12]  Kinzie Jl,et al.  Screening and surveillance for colorectal cancer. , 1988 .

[13]  A. Wyllie,et al.  Apoptosis: A Basic Biological Phenomenon with Wide-ranging Implications in Tissue Kinetics , 1972, British Journal of Cancer.

[14]  H. Kawasaki,et al.  Evidence that Expression of a Mutated p53 Gene Attenuates Apoptotic Cell Death in Human Gastric Intestinal‐type Carcinomas in vivo , 1997, Japanese journal of cancer research : Gann.

[15]  R. Sampliner,et al.  Reduced bile acid-induced apoptosis in "normal" colorectal mucosa: a potential biological marker for cancer risk. , 1996, Cancer research.

[16]  D. Ahlquist,et al.  Morphometric analysis of the "mucocellular layer" overlying colorectal cancer and normal mucosa: relevance to exfoliation and stool screening. , 2000, Human pathology.

[17]  Taylor Murray,et al.  Cancer Statistics, 2001 , 2001, CA: a cancer journal for clinicians.

[18]  D. Henson,et al.  Biomarkers for early detection of colon cancer. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[19]  R. Ward,et al.  Activation of the K‐ras oncogene in colorectal neoplasms is Associated with decreased apoptosis , 1997, Cancer.

[20]  S. Thibodeau,et al.  Colorectal cancer screening by detection of altered human DNA in stool: feasibility of a multitarget assay panel. , 2000, Gastroenterology.

[21]  F. Hamdy.  Prognostic and predictive factors in prostate cancer. , 2001, Cancer treatment reviews.

[22]  S. Goodman,et al.  Detecting colorectal cancer in stool with the use of multiple genetic targets. , 2001, Journal of the National Cancer Institute.

[23]  D. Ahlquist,et al.  Colorectal cancer detection in the practice setting. Impact of fecal blood testing. , 1990, Archives of internal medicine.

[24]  D. Ahlquist,et al.  Stool screening for colorectal cancer: evolution from occult blood to molecular markers. , 2002, Clinica chimica acta; international journal of clinical chemistry.

[25]  D. Ahlquist.  Fecal occult blood testing for colorectal cancer. Can we afford to do this? , 1997, Gastroenterology clinics of North America.

[26]  B. Vogelstein,et al.  A genetic model for colorectal tumorigenesis , 1990, Cell.

[27]  R. Sampliner,et al.  A bile acid-induced apoptosis assay for colon cancer risk and associated quality control studies. , 1999, Cancer research.

[28]  K. Miller,et al.  DNA‐Based Detection of Prostate Cancer in Blood, Urine, and Ejaculates , 2001, Annals of the New York Academy of Sciences.

[29]  H. Bernstein,et al.  Role of apoptosis in biology and pathology: resistance to apoptosis in colon carcinogenesis. , 1995, Ultrastructural pathology.

[30]  S. Schnitt,et al.  Extensive apoptosis in ductal carcinoma in situ of the breast , 1996, Cancer.

[31]  M. Neurath,et al.  Regulation of T-cell apoptosis in inflammatory bowel disease: to die or not to die, that is the mucosal question. , 2001, Trends in immunology.