A gene marker panel covering the Wnt and the Ras-Raf-MEK-MAPK signalling pathways allows to detect gene mutations in 80% of early (UICC I) colon cancer stages in humans

Scholtka, Bettina; Schneider, Mandy; Melcher, Ralph; Katzenberger, Tiemo; Friedrich, D; Berghof-Jäger, Kornelia; Scheppach, Wolfgang; Steinberg, Pablo

doi:10.1016/j.canep.2009.05.001

Cited by 11 publications

(14 citation statements)

References 31 publications

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“…Mutations in APC, KRAS, or BRAF genes have been classified as colorectal cancer-initiating events and thus represent promising biomarkers for molecular detection of beginning cancer and its precursors (2)(3)(4)(5). APC mutations have been described in early cancerous lesions as well as in cancer precursors like adenomas (6,7) and even in aberrant crypt foci (8,9).…”

Section: Introductionmentioning

confidence: 99%

“…From our recent work and from the literature, it is known that different mutation types (insertions, deletions, or base substitutions) can occur in one and the same mutational hotspot, aggravating the detection. For instance, the codon 1309 hotspot which contains repetitive sequences spanning codons 1306-1311, might be affected by a 5 bp deletion leading to a frameshift or by a G>T substitution resulting in a stop codon, whereas the codon 1556 hotspot (codons 1553-1556) most often exhibits a 1 bp insertion of an adenine but can also be affected by a G>T substitution resulting in a stop codon (4,5). For this reason, detection methods using mutant-specific probes are inappropriate in this case and although APC is the most frequently mutated gene in colorectal cancer, there is no sensitive commercial test for APC mutation analysis available up to now.…”

Section: Introductionmentioning

confidence: 99%

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Ultrasensitive Detection of Unknown Colon Cancer-Initiating Mutations Using the Example of the Adenomatous Polyposis Coli Gene

Gerecke

Mascher²,

Gottschalk³

et al. 2013

Cancer Prevention Research

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Detection of cancer precursors contributes to cancer prevention, for example, in the case of colorectal cancer. To record more patients early, ultrasensitive methods are required for the purpose of noninvasive precursor detection in body fluids. Our aim was to develop a method for enrichment and detection of known as well as unknown driver mutations in the Adenomatous polyposis coli (APC) gene. By coupled wild-type blocking (WTB) PCR and high-resolution melting (HRM), referred to as WTB-HRM, a minimum detection limit of 0.01% mutant in excess wild-type was achieved according to as little as 1 pg mutated DNA in the assay. The technique was applied to 80 tissue samples from patients with colorectal cancer (n ¼ 17), adenomas (n ¼ 50), serrated lesions (n ¼ 8), and normal mucosa (n ¼ 5). Any kind of known and unknown APC mutations (deletions, insertions, and base exchanges) being situated inside the mutation cluster region was distinguishable from wild-type DNA. Furthermore, by WTB-HRM, nearly twice as many carcinomas and 1.5 times more precursor lesions were identified to be mutated in APC, as compared with direct sequencing. By analyzing 31 associated stool DNA specimens all but one of the APC mutations could be recovered. Transferability of the WTB-HRM method to other genes was proven using the example of KRAS mutation analysis. In summary, WTB-HRM is a new approach for ultrasensitive detection of cancer-initiating mutations. In this sense, it appears especially applicable for noninvasive detection of colon cancer precursors in body fluids with excess wild-type DNA like stool. Cancer Prev Res; 6(9); 898-907. Ó2013 AACR.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Ultrasensitive Detection of Unknown Colon Cancer-Initiating Mutations Using the Example of the Adenomatous Polyposis Coli Gene

Gerecke

Mascher²,

Gottschalk³

et al. 2013

Cancer Prevention Research

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“…In a previous study on CRC (UICC stages I–IV) [20] B-Raf and CTNNB1 mutations were successfully detected by single strand conformation polymorphism (SSCP) analysis, K-Ras mutations by restriction fragment length polymorphism (RFLP) analysis and APC mutations by direct sequencing of the mutation cluster region. In order to be able to compare the data for the preneoplastic lesions with the results for CRC [20], the same three methods were used.…”

Section: Resultsmentioning

confidence: 99%

“…In order to be able to compare the data for the preneoplastic lesions with the results for CRC [20], the same three methods were used. The results of the gene mutation analysis of all samples included in this study are presented in the Electronic Supplementary Information (Table S1).…”

Section: Resultsmentioning

confidence: 99%

“…These four genes were selected because they are involved in the Wnt and the Ras-Raf-MEK-MAPK signaling cascades and therefore play a substantial role in the adenoma-carcinoma as well as in the serrated adenoma pathway. By making use of this primer panel it could very recently be shown that 74–80% of the analyzed early stage CRC (UICC stages I and II) and over 80% of CRC in UICC stage IV carried at least one of the four above-mentioned genes in a mutated form [20]. …”

Section: Introductionmentioning

confidence: 99%

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Detection of up to 65% of Precancerous Lesions of the Human Colon and Rectum by Mutation Analysis of APC, K-Ras, B-Raf and CTNNB1

Schneider

Scholtka

Gottschalk³

et al. 2010

Cancers

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In the present study a recently conceived 4-gene marker panel covering the Wnt and Ras-Raf-MEK-MAPK signaling pathways was used to analyze 20 colorectal serrated lesions and 41 colorectal adenoma samples and to determine the percentage of each of the above-mentioned potentially precancerous lesions carrying at least one of the four above-mentioned genes in a mutated form. CTNNB1 and B-Raf were screened by PCR-single-strand conformation polymorphism analysis, K-Ras by restriction fragment length polymorphism analysis and the APC gene mutation cluster region (codons 1243–1567) by direct DNA sequencing. APC mutations were only detected in 10% of the serrated lesions but in 34% of the adenomas. Twenty percent of the serrated lesions and 14% of the adenomas carried a mutated K-Ras. B-Raf was found to be mutated in 50% of the serrated lesions and in 22% of the adenomas. CTNNB1 was altered in 12% of the adenomas, but not in serrated lesions. By using the above gene marker panel it could be shown that 65% of the serrated lesions and 61% of the adenomas carried at least one of the four genes in a mutated form. Based on its excellent performance in detecting mutations in sporadic preneoplastic (in this study) and neoplastic lesions (in a previous study) of the human colon and rectum, this primer combination might also be suited to efficiently and non-invasively detect genetic alterations in stool DNA of patients with early colorectal cancer.

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Colorectal cancers choosing sides

Albuquerque¹,

Bakker

Veelen

et al. 2011

Biochimica et Biophysica Acta (BBA) - Reviews on Cancer

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In contrast to the majority of sporadic colorectal cancer which predominantly occur in the distal colon, most mismatch repair deficient tumours arise at the proximal side. At present, these regional preferences have not been explained properly. Recently, we have screened colorectal tumours for mutations in Wnt-related genes focusing specifically on colorectal location. Combining this analysis with published data, we propose a mechanism underlying the side-related preferences of colorectal cancers, based on the specific acquired genetic defects in β-catenin signalling.

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A gene marker panel covering the Wnt and the Ras-Raf-MEK-MAPK signalling pathways allows to detect gene mutations in 80% of early (UICC I) colon cancer stages in humans

Abstract: A gene marker panel covering the Wnt and the Ras-Raf-MEK-MAPK signalling pathways allows to detect gene mutations in 80% of early (UICC I) colon cancer stages in humans first published in:

Cited by 11 publications

References 31 publications

Ultrasensitive Detection of Unknown Colon Cancer-Initiating Mutations Using the Example of the Adenomatous Polyposis Coli Gene

Ultrasensitive Detection of Unknown Colon Cancer-Initiating Mutations Using the Example of the Adenomatous Polyposis Coli Gene

Detection of up to 65% of Precancerous Lesions of the Human Colon and Rectum by Mutation Analysis of APC, K-Ras, B-Raf and CTNNB1

Colorectal cancers choosing sides

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