Cancer Research AACR Legacy Telomeres
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online
This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Findeisen, P.
Right arrow Articles by von Knebel Doeberitz, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Findeisen, P.
Right arrow Articles by von Knebel Doeberitz, M.
[Cancer Research 65, 8072-8078, September 15, 2005]
© 2005 American Association for Cancer Research

Priority Reports

T25 Repeat in the 3' Untranslated Region of the CASP2 Gene: A Sensitive and Specific Marker for Microsatellite Instability in Colorectal Cancer

Peter Findeisen1,2, Matthias Kloor1, Sabine Merx1,2, Christian Sutter1,3, Stefan M. Woerner1, Nicole Dostmann1, Axel Benner4, Bolormaa Dondog5, Michael Pawlita5, Wolfgang Dippold6, Rudolf Wagner7, Johannes Gebert1 and Magnus von Knebel Doeberitz1

1 Department of Pathology, Institute of Molecular Pathology; 2 Institute of Clinical Chemistry, Medical Faculty Mannheim; 3 Department of Human Genetics, Institute of Human Genetics, University of Heidelberg; 4 Central Unit Biostatistics, 5 Research Program Infection and Cancer, German Cancer Research Center, Heidelberg, Germany; 6 St. Vincenz und Elisabeth-Hospital, Mainz, Germany; and 7 Institute of Pathology, Westpfalz Klinikum, Kaiserslautern, Germany

Requests for reprints: Magnus von Knebel Doeberitz, Department of Pathology, Institute of Molecular Pathology, University of Heidelberg, Im Neuenheimer Feld 220, D-69120 Heidelberg, Germany. Phone: 49-6221-56-2876; Fax: 49-6221-56-5981; E-mail: knebel{at}med.uni-heidelberg.de and mvkd{at}aol.com.


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
DNA mismatch repair deficiency is observed in about 10% to 15% of all colorectal carcinomas and in up to 90% of hereditary nonpolyposis colorectal cancer (HNPCC) patients. Tumors with mismatch repair defects acquire mutations in short repetitive DNA sequences, a phenomenon termed high-level microsatellite instability (MSI-H). The diagnosis of MSI-H in colon cancer is of increasing relevance, because MSI-H is an independent prognostic factor in colorectal cancer, seems to influence the efficacy of adjuvant chemotherapy, and is the most important molecular screening tool to identify HNPCC patients. To make MSI typing feasible for the routine pathology laboratory, highly reproducible and cost effective laboratory tests are required. Here, we describe a novel T25 mononucleotide marker in the 3'untranslated region of the CASP2 gene (CAT25) that displayed a quasimonomorphic repeat pattern in normal tissue of 200 unrelated individuals of Caucasian origin. In addition, CAT25 was monomorphic also in all tested donors of African and Asian origin (n = 102 and n = 79, respectively) and thus differs from the most commonly used markers BAT25 and BAT26. Without the analysis of corresponding normal tissue, CAT25 correctly detected 56 of 57 colorectal cancer specimens classified as MSI-H by using the standard National Cancer Institute/International Collaborative Group-HNPCC marker panel. Combined with the standard markers BAT25 and BAT26 in a multiplex PCR, all MSI-H colorectal cancer samples were typed correctly. No false-positive results were obtained in 60 non-MSI-H control colorectal cancer specimens. These data suggest that CAT25 should be included into novel marker panels for microsatellite testing thus allowing for a significant reduction of the complexity and costs of MSI typing. Moreover, CAT25 represents a highly promising marker for early detection of colorectal cancer in HNPCC germ line mutation carriers.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
Microsatellite instability (MSI) is observed in about 10% to 15% of sporadic colorectal carcinomas and the majority of those occurring in hereditary nonpolyposis colorectal cancer (HNPCC) patients that harbor germ line mutations in DNA mismatch repair genes (for a review, see ref. 1). Colorectal cancers displaying a high level of MSI (MSI-H) possess particular pathologic and clinical features. MSI-H colorectal cancers are often localized in the proximal colon and present with a dense intratumoral lymphocyte infiltration (2). Several studies suggest that patients with MSI-H colorectal cancers have a better prognosis (e.g., refs. 3, 4). Most importantly, recent studies indicate that benefit from adjuvant 5-fluorouracil chemotherapy may be restricted to non-MSI-H colorectal cancer patients (5, 6). MSI typing is therefore likely to become a routine diagnostic procedure in the pathology laboratory. However, at present, it is usually only applied to patients preselected upon clinical criteria (Bethesda guidelines; refs. 7, 8), because the standard testing procedure recommended by the National Cancer Institute/International Collaborative Group/HNPCC (NCI/ICG-HNPCC; ref. 7) implies a considerable laboratory workload. That is, five microsatellite markers including two mononucleotide repeats (BAT26 and BAT25) and three dinucleotide repeats (D2S123, D5S346, and D17S250) have to be amplified from DNA of tumor and normal tissue. A panel of five additional MSI markers is used for MSI classification of borderline cases. Apart from an overall requirement for several diagnostic markers and often limited amounts of available tissues, analysis of matched normal DNA of the same patient make MSI analysis a laborious and costly procedure that is not applicable for high throughput screening.

The aim of the present study was to establish a novel and simple MSI test with similar or even higher sensitivity compared with the current reference panel. As we describe here, amplification of the 3'untranslated region (3'UTR) T25 mononucleotide repeat of the Caspase 2 gene (CASP2, referred to as CAT25) fulfills the demands of a simple and reliable MSI-typing procedure. Only one PCR amplifying CAT25 alone or multiplexed with BAT25 and BAT26 from tumor DNA is sufficient to yield the same sensitivity and specificity as the five-marker panel recommended by the NCI/ICG-HNPCC.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Sample preparation and DNA extraction. Genomic DNA was isolated from formalin-fixed, paraffin-embedded colorectal cancer specimens (n = 117) after manual microdissection using a commercially available kit (DNeasy Tissue KIT, Qiagen, Hilden, Germany). For the evaluation of CAT25 allele distribution, genomic DNA was isolated from nondiseased tissue samples of unrelated Caucasian (n = 200), African (n = 102), and Asian (n = 79) individuals. African DNA samples have been described previously (9). Asian DNA samples were isolated from healthy individuals participating in an ongoing human papillomavirus prevalence study in Ulaanbaatar, Mongolia.

Microsatellite instability analysis and multiplex PCR. Typing of MSI was carried out using the standard NCI/ICG-HNPCC marker panel (7) as described previously (10). Tumors were classified as MSI-H if at least 30% of the markers displayed instability; low microsatellite unstable (MSI-L) with instability in <30% of the markers or microsatellite stable (MSS) if no instabilities were detected. PCR primers for the amplification of CAT25 (forward 5'-CCTAGAAACCTTTATCCCTGCTT-3' and reverse 5'-GAGCTTGCAGTGAGCTGAGA-3') were designed using the "Primer3" software (Whitehead MIT, Cambridge, MA).8 Sense PCR primers were labeled at the 5' end with FITC (BAT26 and CAT25) or HEX fluorescent dye (BAT25), respectively. Multiplex PCR was carried out in a total reaction volume of 25 µL using a final concentration of 200 µmol/L deoxynucleotide triphosphates, 12.5 pmol/L of each primer, 1x PCR buffer [20 mmol/L Tris-HCl (pH 8.4), 50 mmol/L KCl], 1.5 mmol/L MgCl2, and 0.75 units of Taq DNA polymerase (Life Technologies/BRL, Eggenstein, Germany). Genomic DNA (50 ng) was used as a template. Reaction mixes were subjected to the following conditions: initial denaturation at 94°C for 5 minutes followed by 38 cycles of denaturation at 94°C for 30 seconds, annealing at 55°C for 30 seconds, extension at 72°C for 30 seconds, and a final extension step at 72°C for 7 minutes.

Fragment analysis and statistical evaluation. For fragment analysis, 2 µL of appropriately diluted PCR products were mixed with 12 µL of formamide and 0.2 µL ROX500 length standard (Applied Biosystems, Darmstadt, Germany). DNA fragments were separated on ABI PRISM 3100 Genetic Analyzer (Applied Biosystems) using Filter Set D. Size, height, and profiles of microsatellite peaks were analyzed using the GeneScan 3.7 software (Applied Biosystems). The electropherogram peak with the largest area was defined as "main product". For each tumor, the number of shifted bases in comparison with the corresponding normal mucosa was counted, and mean basepair deletions were calculated for each mononucleotide microsatellite marker. Statistical analysis and graphical displays were done by using the Statistica 6.0 software (StatSoft, Tulsa, OK). For comparison of diagnostic sensitivity/specificity of the markers CAT25, BAT25, and BAT26, ROC analysis was done. Areas under the ROC curve were compared according to the method described by DeLong et al. (11).


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Allelic size distribution of CAT25 in colorectal cancer and nontumorous tissue specimens. During our search for microsatellites located in 3' UTRs of cancer-relevant genes, we identified a T25 repeat (CAT25) located within the 3'UTR of the CASP2 gene (Genbank accession no. NM_032982; position, nucleotides 2685-2709) that displayed a monomorphic pattern in a small subset of normal tissue samples and 100% mutation frequency in 10 MSI-H colorectal cancer specimens. These results suggested to us that CAT25 may be a suitable marker for MSI testing, potentially without the need of corresponding normal tissue. To examine the diagnostic sensitivity and specificity of CAT25 for MSI analysis in more detail, analysis of CAT25 was extended to a larger number of normal tissue, MSI-H, and non-MSI-H colorectal cancer specimens.

Because MSI analysis of tumors in the absence of corresponding normal tissue requires markers with a monomorphic or quasimonomorphic allele distribution, we first examined whether CAT25 fulfilled this criterion in normal DNA samples from unrelated individuals of Caucasian origin (n = 200). The PCR product most frequently observed (128 individuals, 64.0%) was 147 bp in length (referred to as "wild-type allele"), a product of 146 bp was detected in 18 cases (9.0%), and a product of 148 bp in 54 cases (27.0%). No additional alleles shorter than 146 bp or longer than 148 bp in length were observed. For BAT25 and BAT26, considerable allelic size variation has been described in the African population. To analyze the allelic profile of CAT25 in populations of different ethnic origin, the CAT25 locus was amplified from normal tissue specimens from African (n = 102) and Asian (n = 79) individuals. In contrast to BAT25 and BAT26 which exhibited shorter alleles potentially causing misclassification of MSI in 26.5% and 21.6% of the tested African individuals, CAT25 was proved to be monomorphic in all tested individuals of African, Asian, and Caucasian origin (Table 1). To exclude single nucleotide polymorphisms at the CAT25 locus in African individuals, which might affect diagnostic sensitivity of the marker in this population, sequencing of CAT25 PCR products amplified from African donors (n = 20) was done. No polymorphisms were observed (data not shown). Based on the allelic profile of CAT25, we defined any product length of CAT25 <146 or >148 bp as indicative of MSI.


View this table:
[in this window]
[in a new window]

  		Table 1. Allele length distribution of BAT25, BAT26, and CAT25

 
As the next step, colorectal cancer specimens previously typed for their microsatellite status using the standard NCI/ICG-HNPCC marker panel (7) as MSI-H (n = 57) or non-MSI-H (MSI-L, n = 10; MSS, n = 50) were analyzed for mutations at the CAT25 locus. No CAT25 mutations were observed in any of the MSS or MSI-L colorectal cancer samples (data not shown). However, in 57 of 57 (100%) MSI-H colorectal cancers, mutations at the CAT25 locus were detected (Table 2). However, without the analysis of matched normal tissue DNA, only 56 of 57 (98.2%) MSI-H colorectal cancers would have been classified correctly by CAT25, because in one case, the product amplified from the mutated allele had an allele length of 146 bp, which had also been observed in 9% of normal tissue specimens (Fig. 1, bottom).


View this table:
[in this window]
[in a new window]

  		Table 2. Instability profile of ICG-HNPCC microsatellite markers and CAT25 in 57 MSI-H colorectal cancers

 


View larger version (28K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 1. Distribution of BAT25, BAT26, and CAT25 product lengths in tumor and normal tissue specimens from Caucasian individuals. Relative product lengths observed in MSI-H CRCs (shaded columns, n = 57) and nontumorous control DNA samples (dashed columns, n = 57 for BAT25 and BAT26, n = 200 for CAT25). Without the analysis of corresponding normal tissue, MSI-H was not detected in one case for CAT25, in one case for BAT25, and in two cases for BAT26. {04-4146fx04l}, MSI-H CRC; {blacksquare}, normal.

 
For comparison of CAT25 with the established reference markers BAT25 and BAT26, ROC analysis was done to assess the diagnostic accuracy of these markers when used alone for the diagnosis of MSI in Caucasian individuals. ROC analysis revealed an area under curve (AUC) of 0.9996 for CAT25 compared with 0.9914 and 0.995 for BAT25 and BAT26, respectively. Although there was a trend towards a higher diagnostic accuracy of the CAT25 marker, differences were not statistically significant. Similar AUC values were obtained when combining MSI-H colorectal cancer data with normal alleles from the Asian population (data not shown). However, when comparing alleles amplified from MSI-H tumor tissue with normal alleles from African donors, hypothetical ROC analysis revealed AUC values of 0.9997 for CAT25, 0.9028 for BAT25, and 0.8785 for BAT26. Differences between diagnostic accuracy of CAT25 and BAT25/BAT26 were highly significant (P < 0.0001). This indicates that in contrast to BAT25 and BAT26, CAT25 may be used for MSI analysis without amplification of normal tissue also in African individuals.

The MSI-H phenotype is detected more easily if it is accompanied by large deletions and/or insertions at the microsatellite loci used for MSI typing. For BAT26, average ± SD basepair deletion was highest with 8.9 ± 3.1 bp followed by CAT25 (8.1 ± 2.9 bp) and BAT25 (6.0 ± 2.2 bp). Accordingly, the CAT25 marker ranged between the mononucleotide markers BAT26 and BAT25 in terms of the average size of the basepair deletion. Product length distribution of BAT25 and BAT26 in MSI-H colorectal cancers and corresponding nontumorous tissues are depicted in Fig. 1 (top and middle).

Combination of CAT25, BAT25, and BAT26 in a multiplex PCR. Our results predicted that in Caucasians, a multiplex protocol that combines CAT25 with the most sensitive markers of the ICG-HNPCC reference panel (BAT25 and BAT26) in one triplex PCR should detect the MSI-H phenotype with maximal sensitivity and specificity, even without concomitant analysis of normal tissue. By using this combinatorial strategy (BAT25/BAT26/CAT25), the product lengths previously obtained in single PCRs were confirmed by this triplex approach without exception. Most importantly, 117 of 117 (100%) colorectal cancer samples were classified correctly as MSI-H or non-MSI-H. In each MSI-H tumor, at least two of the three mononucleotide markers were found indicative of MSI (Table 2). Examples of fragment patterns obtained by this multiplex approach are presented in Fig. 2.



View larger version (33K):
[in this window]
[in a new window]
[Download PPT slide]
  		Figure 2. Multiplex PCR amplification of BAT26, BAT25, and CAT25 from two MSI-H colorectal cancers (A and B), one MSS colorectal cancer (C), and one MSI-L colorectal cancer (D). PCR products are displayed for BAT26 (blue; average relative product length, 117 bp), BAT25 (green, 122 bp), and CAT25 (blue, 147 bp). Red peaks represent the internal length standard. Filled peaks span the largest area and are defined as main products. The range of product lengths observed in normal DNA of healthy Caucasian individuals is indicated by dashed lines for each marker. Shift lengths are denoted above the corresponding product peaks.

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
Recent findings concerning the characteristic clinical behavior of MSI-H colorectal cancers suggest routine MSI screening for all newly diagnosed colorectal cancers. The current standard method is comparatively time-consuming, laborious, expensive, and also often remains ambiguous due to lack of matched normal tissues. Therefore, a simplified strategy is required to render MSI analysis feasible for high throughput application.

To reduce the workload of MSI testing, several techniques have been suggested previously. Immunohistochemistry with monoclonal antibodies specific for MLH1 and MSH2 is commonly accepted as a useful lower-cost tool to identify MSI-H colorectal cancers; however, about 10% of MSI-H tumors are missed if immunohistochemistry is used alone (12). Hence, we believe that PCR-based MSI testing is indispensable for correct MSI classification at present, at least in tumors without abnormal immunohistochemical staining results (13).

The aim of this study was the identification and characterization of an MSI test system attaining similar sensitivity as the NCI/ICG-HNPCC standard marker panel and requiring only one single PCR without the need for corresponding normal tissue. The use of BAT26 alone has been suggested for this purpose, even without the need for matching normal tissue (14, 15). Although this approach may be sufficient for the majority of MSI-H cases, it remains less sensitive than the NCI/ICG-HNPCC standard panel (16). For example, in our set of MSI-H tumors, two cases would have been missed by using BAT26 as single diagnostic MSI marker. Additionally, depending on the ethnic origin of the tested individuals, shortened BAT26 and BAT25 alleles which have been reported in up to 5.3% and up to 6.8%, respectively (17–19), may lead to false-positive results, rendering the interpretation of MSI analysis difficult if it is based on amplification of BAT26 and BAT25 from tumor tissue alone. In our control samples from different populations, about 21.6% and 26.5% of individuals from African origin displayed shorter BAT25 and BAT26 alleles, respectively.

Therefore, we recommend the inclusion of novel markers to overcome these limitations. The ideal marker for MSI analysis should match the following criteria: (i) 100% mutation frequency in the MSI-H tumors of interest, (ii) no mutations in MSS tissue specimens, and (iii) a (quasi)monomorphic allele pattern in all populations. Our data show that CAT25 fulfills all these criteria. CAT25 mutations were detected in all MSI-H colorectal cancers, 56 of 57 (98.2%) were indicative of MSI without the analysis of normal tissue. In contrast, no CAT25 alterations were observed in 50 control MSS or 10 MSI-L colorectal cancer samples. Finally, a quasimonomorphic pattern was detected in 381 individuals from different ethnic origin. These data suggest that CAT25 may also be a highly promising marker for early detection of MSI-H colorectal cancer in HNPCC germ line mutation carriers, potentially surpassing BAT26, a marker previously suggested for this application (20), in sensitivity and specificity, particularly when applied to individuals of African origin.

In comparison with a previously suggested pentaplex set containing the mononucleotide markers NR-21, NR-22, and NR-24 (21), or additional mononucleotide markers like MONO-27 and BAT-34c4 (22), CAT25 seems to possess a higher sensitivity or specificity for detecting the MSI-H phenotype in colorectal cancer. In addition, judgment of CAT25 profiles should be easier compared with these markers, because the average basepair deletion in MSI-H colorectal cancers is markedly more pronounced at the CAT25 locus. This may be of relevance for the development of future automated test systems. By combination of CAT25, BAT25, and BAT26 in one triplex PCR, all tumors in the present study were typed correctly for their MSI status. To minimize false-positive results due to allelic size variations at the BAT25 or BAT26 locus, we suggest positive scoring only if at least two of the triplex markers present with mutations.

In accordance with recent suggestions (23), we recommend the combined use of quasimonomorphic mononucleotide repeats for MSI testing. It is widely accepted that additional mononucleotide markers have to be evaluated for this purpose (8). In this study, we report a novel mononucleotide marker, CAT25, that represents a highly promising candidate marker for future high-throughput MSI testing and also for early detection approaches. To achieve 100% diagnostic sensitivity and 100% specificity in the Caucasian population, we recommend a triplex PCR of CAT25, BAT25, and BAT26. To the best of our knowledge, this combination represents the simplest maximum sensitivity MSI detection method reported thus far. For the development of a test system displaying maximal accuracy in all populations, combination of CAT25 with alternative markers needs to be evaluated. Systematic studies analyzing the mutation frequency of CAT25 in a large cohort of patients with colorectal cancer or extracolonic malignancies are currently in progress.


   Acknowledgments
 
Grant support: German Cancer Aid Deutsche Krebshilfe grant 70-3026-Kn 5.

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

We thank the excellent technical assistance of B. Kuchenbuch and I. Voehringer.


   Footnotes
 
Note: P. Findeisen, M. Kloor, and S. Merx contributed equally to this work.

8 http://www-genome.wi.mit.edu/genome_software/other/primer3.html. Back

Received 11/18/04. Revised 5/ 1/05. Accepted 7/19/05.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Umar A, Risinger JI, Hawk ET, Barrett JC. Testing guidelines for hereditary non-polyposis colorectal cancer. Nat Rev Cancer 2004;4:153–8.[Medline]
  2. Smyrk TC, Watson P, Kaul K, Lynch HT. Tumor-infiltrating lymphocytes are a marker for microsatellite instability in colorectal carcinoma. Cancer 2001;91:2417–22.[CrossRef][Medline]
  3. Gryfe R, Kim H, Hsieh ET, et al. Tumor microsatellite instability and clinical outcome in young patients with colorectal cancer. N Engl J Med 2000;342:69–77.[Abstract/Free Full Text]
  4. Samowitz WS, Curtin K, Ma KN, et al. Microsatellite instability in sporadic colon cancer is associated with an improved prognosis at the population level. Cancer Epidemiol Biomarkers Prev 2001;10:917–23.[Abstract/Free Full Text]
  5. Ribic CM, Sargent DJ, Moore MJ, et al. Tumor microsatellite instability status as a predictor of benefit from fluorouracil-based adjuvant chemotherapy for colon cancer. N Engl J Med 2003;349:247–57.[Abstract/Free Full Text]
  6. Carethers JM, Smith EJ, Behling CA, et al. Use of 5-fluorouracil and survival in patients with microsatellite-unstable colorectal cancer. Gastroenterology 2004;126:394–401.[CrossRef][Medline]
  7. Boland CR, Thibodeau SN, Hamilton SR, et al. A National Cancer Institute workshop on microsatellite instability for cancer detection and familial predisposition: development of international criteria for the determination of microsatellite instability in colon cancer. Cancer Res 1998;58:5248–57.[Abstract/Free Full Text]
  8. Umar A, Boland CR, Terdiman JP, et al. Revised Bethesda Guidelines for hereditary nonpolyposis colorectal cancer (Lynch syndrome) and microsatellite instability. J Natl Cancer Inst 2004;96:261–8.[Abstract/Free Full Text]
  9. ter Meulen J, Eberhardt HC, Luande J, et al. Human papillomavirus (HPV) infection, HIV infection and cervical cancer in Tanzania, east Africa. Int J Cancer 1992;51:515–21.[Medline]
  10. Sutter C, Gebert J, Bischoff P, Herfarth C, von Knebel Doeberitz M. Molecular screening of potential HNPCC patients using a multiplex microsatellite PCR system. Mol Cell Probes 1999;13:157–65.[CrossRef][Medline]
  11. DeLong E, DeLong D, Clarke-Pearson D. Comparing the areas under two or more correlated receiver operating characteristic curves: a nonparametric approach. Biometrics 1988;44:837–45.[CrossRef][Medline]
  12. Lindor NM, Burgart LJ, Leontovich O, et al. Immunohistochemistry versus microsatellite instability testing in phenotyping colorectal tumors. J Clin Oncol 2002;20:1043–8.[Abstract/Free Full Text]
  13. Engel C, Forberg J, Holinski-Feder E, et al., German HNPCC consortium. A novel strategy for optimal sequential application of clinical criteria, immunohistochemistry and microsatellite analysis in the diagnosis of hereditary nonpolyposis colorectal cancer. Int J Cancer. Epub 2005 Jul 7.
  14. Hoang JM, Cottu PH, Thuille B, Salmon RJ, Thomas G, Hamelin R. BAT-26, an indicator of the replication error phenotype in colorectal cancers and cell lines. Cancer Res 1997;57:300–3.[Abstract/Free Full Text]
  15. Zhou XP, Hoang JM, Li YJ, et al. Determination of the replication error phenotype in human tumors without the requirement for matching normal DNA by analysis of mononucleotide repeat microsatellites. Genes Chromosomes Cancer 1998;21:101–7.[CrossRef][Medline]
  16. Perucho M. Correspondence re: C.R. Boland et al., A National Cancer Institute workshop on microsatellite instability for cancer detection and familial predisposition: development of international criteria for the determination of microsatellite instability in colorectal cancer, Cancer Res 1998;58:5248–57. Cancer Res 1999;59:249–56.[Free Full Text]
  17. Zhou XP, Hoang JM, Cottu P, Thomas G, Hamelin R. Allelic profiles of mononucleotide repeat microsatellites in control individuals and in colorectal tumors with and without replication errors. Oncogene 1997;15:1713–8.[CrossRef][Medline]
  18. Pyatt R, Chadwick RB, Johnson CK, Adebamowo C, de la Chapelle A, Prior TW. Polymorphic variation at the BAT-25 and BAT-26 loci in individuals of African origin. Implications for microsatellite instability testing. Am J Pathol 1999;155:349–53.[Abstract/Free Full Text]
  19. Ichikawa A, Sugano K, Fujita S. DNA variants of BAT-25 in Japanese, a locus frequently used for analysis of microsatellite instability. Jpn J Clin Oncol 2001;31:346–8.[Abstract/Free Full Text]
  20. Traverso G, Shuber A, Olsson L, et al. Detection of proximal colorectal cancers through analysis of faecal DNA. Lancet 2002;359:403–4.[CrossRef][Medline]
  21. Suraweera N, Duval A, Reperant M, et al. Evaluation of tumor microsatellite instability using five quasimonomorphic mononucleotide repeats and pentaplex PCR. Gastroenterology 2002;12:1804–11.[CrossRef]
  22. Bacher JW, Flanagan LA, Smalley RL, et al. Development of a fluorescent multiplex assay for detection of MSI-High tumors. Dis Markers 2004;20:237–50.[Medline]
  23. Buhard O, Suraweera N, Lectard A, Duval A, Hamelin R. Quasimonomorphic mononucleotide repeats for high-level microsatellite instability analysis. Dis Markers 2004;20:251–7.[Medline]



This article has been cited by other articles:


Home page
 J. Mol. Diagn.Home page
F. Bianchi, E. Galizia, R. Catalani, L. Belvederesi, C. Ferretti, F. Corradini, and R. Cellerino
CAT25 Is a Mononucleotide Marker to Identify HNPCC Patients
J. Mol. Diagn., May 1, 2009; 11(3): 248 - 252.
[Abstract] [Full Text] [PDF]

Home page
 Cancer Res.Home page
N. Roeckel, S. M. Woerner, M. Kloor, Y.-P. Yuan, G. Patsos, R. Gromes, J. Kopitz, and J. Gebert
High Frequency of LMAN1 Abnormalities in Colorectal Tumors with Microsatellite Instability
Cancer Res., January 1, 2009; 69(1): 292 - 299.
[Abstract] [Full Text] [PDF]

Home page
 CarcinogenesisHome page
K. Imai and H. Yamamoto
Carcinogenesis and microsatellite instability: the interrelationship between genetics and epigenetics
Carcinogenesis, April 1, 2008; 29(4): 673 - 680.
[Abstract] [Full Text] [PDF]

Home page
 J. Mol. Diagn.Home page
V. Deschoolmeester, M. Baay, W. Wuyts, E. Van Marck, N. Van Damme, P. Vermeulen, K. Lukaszuk, F. Lardon, and J. B. Vermorken
Detection of Microsatellite Instability in Colorectal Cancer Using an Alternative Multiplex Assay of Quasi-Monomorphic Mononucleotide Markers
J. Mol. Diagn., March 1, 2008; 10(2): 154 - 159.
[Abstract] [Full Text] [PDF]

This Article
Right arrow Abstract Freely available
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Findeisen, P.
Right arrow Articles by von Knebel Doeberitz, M.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Findeisen, P.
Right arrow Articles by von Knebel Doeberitz, M.

HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Cancer Research Clinical Cancer Research
Cancer Epidemiology Biomarkers & Prevention Molecular Cancer Therapeutics
Molecular Cancer Research Cancer Prevention Research
Cancer Prevention Journals Portal Cancer Reviews Online
Annual Meeting Education Book Meeting Abstracts Online