Utilization ofK-ras mutations identified in stool DNA for the early detection of colorectal cancer

Lev, Ze'ev; Kislitsin, D.; Rennert, Gad; Lerner, Aaron

doi:10.1002/(sici)1097-4644(2000)77:34+<35::aid-jcb8>3.0.co;2-w

Cited by 27 publications

(8 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Subsequent reports also involved interrogation of single genetic targets. [13][14][15][16][17] More recently, assays with multiple markers have been developed 18,19 to yield increased assay sensitivity in light of the genetic heterogeneity of sporadic CRC cases. Furthermore, an assay for long DNA recovered from stool has been developed and shown to be associated with CRC with high specificity.…”

Section: Discussionmentioning

confidence: 99%

DNA Stabilization Is Critical for Maximizing Performance of Fecal DNA-Based Colorectal Cancer Tests

Olson

Whitney²,

Durkee³

et al. 2005

Diagnostic Molecular Pathology

View full text Add to dashboard Cite

We have developed a multitarget, fecal DNA screening assay that detects the presence of gene-specific mutations and long DNA fragments associated with colorectal cancer (CRC). We continue to investigate methods that may be used to optimize clinical sensitivity. The goals of this investigation are to establish how sample handling conditions affect the stability of DNA in stool, thereby potentially limiting clinical sensitivity, and to determine conditions to ameliorate DNA degradation. A study was run comparing paired sample aliquots. Quantitative PCR data for matched aliquots was used to determine first the effect of sample incubation on total recovery and integrity of DNA, then the effect of stabilization buffer addition to stool on recoverable DNA, and finally the impact of buffer addition on assay sensitivity. Comparison of quantitative PCR data for paired aliquots shows that the amount of recoverable human DNA is negatively affected by storing stool samples (N = 43) at room temperature for > or =36 hours (P = 0.0018). However, the addition of stabilization buffer leads to a significant increase in recovery of DNA (P = 0.010), compared with samples incubated without buffer. Whereas the DNA Integrity Assay (DIA) is found to be sensitive to DNA degradation (sensitivity was reduced by 82%; P = 0.0002), point mutation marker sensitivity is more refractory. Overall, DNA can be stabilized by addition of buffer to the sample, leading to increased assay sensitivity.

show abstract

Section: Discussionmentioning

confidence: 99%

DNA Stabilization Is Critical for Maximizing Performance of Fecal DNA-Based Colorectal Cancer Tests

Olson

Whitney²,

Durkee³

et al. 2005

Diagnostic Molecular Pathology

View full text Add to dashboard Cite

show abstract

“…An important step in the progression of colorectal cancer is the induction of activating mutations in KRAS proto-oncogene, GTPase (KRAS). Mutations in KRAS appear in the intermediate adenoma stage, early during tumorigenesis, and it is thus possible to use them as a biomarker for early detection of ~40% of colorectal tumors (8).…”

Section: Introductionmentioning

confidence: 99%

Coffee reduces KRAS expression in Caco-2 human colon carcinoma cells via regulation of miRNAs

2017

View full text Add to dashboard Cite

Abstract.Previous epidemiological studies have demonstrated that moderate coffee consumption is associated with a lower risk of certain types of cancer, particularly colon cancer. To elucidate the molecular basis for this protective action, the effect of coffee on Caco-2 human colon carcinoma cells was investigated. Low concentrations of coffee (<5%) inhibited proliferation of Caco-2 cells without affecting cell viability. Coffee also reduced KRAS proto-oncogene, GTPase (KRAS) gene expression in a dose-dependent manner; however, caffeine, caffeic acid and chlorogenic acid, three major constituents of coffee, did not exhibit this effect. Increasing the duration of coffee bean roasting increased the reduction in KRAS expression, suggesting that the active constituents responsible for this effect emerged during the roasting process. MicroRNA (miR) analysis revealed that coffee induced the expression of miR-30c and miR-96, both of which target the KRAS gene. The results of the present study suggested that daily coffee consumption may reduce KRAS activity, thereby preventing the malignant growth of colon carcinoma cells.

show abstract

“…Currently, the majority of studies have shown that mutations are found more frequently in tissues than in stools. 13,29,30,35,36 In this study, four patients (nos. 2, 17, 20, and 26) exhibiting mutant electrophoresis patterns in both stool samples and cancer tissues were found by direct sequencing to carry K-ras mutations in their cancer tissues but not in their stool samples, whereas clone sequencing confirmed the K-ras mutations in their stool samples, showing that the proportions of mutant K-ras were 9, 12, 15, and 13%, respectively, which demonstrated the lower abundance of K-ras mutations in stool samples compared with that in cancer tissues.…”

Section: Discussionmentioning

confidence: 98%

“…[4][5][6] Based on the good clinical compliance and the findings of several research groups that revealed K-ras mutations in stools consistent with the K-ras status in matching Chip-based detection of stool K-ras mutation H Zhang et al tumor tissues, [28][29][30] mutated DNA in stools has become the latest target of several detection methods. [8][9][10] Although stoolbased DNA detection could report CRC tissue K-ras genotypes faithfully, severe difficulties because of complicated procedures and low sensitivity have hampered further progress in using stool samples to detect K-ras mutations and preventatively screen for CRC.…”

Section: Discussionmentioning

confidence: 99%

Rapid detection of low-abundance K-ras mutation in stools of colorectal cancer patients using chip-based temperature gradient capillary electrophoresis

Zhang

Wang

et al. 2011

Laboratory Investigation

View full text Add to dashboard Cite

Mutant K-ras provides an independent negative predictive marker for epidermal growth factor receptor (EGFR)-targeted therapy in colorectal cancers (CRCs). Rapid, sensitive, and cost-effective screening for K-ras status will overarch rational personalized medicine. Stool-based DNA testing offers unique advantages for CRC screening such as noninvasiveness, high specificity, and patient compliance, whereas complicated procedures and the low sensitivity of the present approaches have hampered its application on a wide scale. In this study, a chip-based temperature gradient capillary electrophoresis (TGCE) technique was applied to detect low-abundance K-ras mutations under a pooled experiment and analyze K-ras mutations in 30 paired stool samples and cancer tissues of CRC patients and 15 stool samples of healthy volunteers. The chip-based TGCE results showed that the successful analysis of K-ras status could be achieved within 6 min with an extremely low sample consumption of 14 nl. Detection is sensitive enough to reliably report 0.2% mutant CRC cells in a wild-type background, and 0.5 ng of template DNA was sufficient for chip-based TGCE. Of the 30 stool samples of CRC patients analyzed, 17 (57%) harbored K-ras mutations, and the lowest percentage of the detectable mutant K-ras in stool samples was 2%. The coincidence rate for K-ras mutations between stools and tissues obtained by the chip-based method reached 97% (29/30). One of the 15 stool samples of normal controls carried K-ras mutations, producing a specificity of 93%. Clone sequencing data entirely confirmed the results obtained by chip-based TGCE. The study demonstrates that chip-based TGCE is capable of rapidly screening low-abundance K-ras mutations with high sensitivity, reproducibility, simplicity, and significant savings of time and sample. Application of this method to genotype the K-ras gene in stools would provide a potential means for predicting the effectiveness of EGFR-targeted therapy in CRC patients using noninvasive approaches.

show abstract

Utilization ofK-ras mutations identified in stool DNA for the early detection of colorectal cancer

Cited by 27 publications

References 27 publications

DNA Stabilization Is Critical for Maximizing Performance of Fecal DNA-Based Colorectal Cancer Tests

DNA Stabilization Is Critical for Maximizing Performance of Fecal DNA-Based Colorectal Cancer Tests

Coffee reduces KRAS expression in Caco-2 human colon carcinoma cells via regulation of miRNAs

Rapid detection of low-abundance K-ras mutation in stools of colorectal cancer patients using chip-based temperature gradient capillary electrophoresis

Contact Info

Product

Resources

About