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Review
. 2002 Apr;55(2):73-9.
doi: 10.1136/mp.55.2.73.

Methods of molecular analysis: mutation detection in solid tumours

I M Frayling  1
Affiliations
Review

Methods of molecular analysis: mutation detection in solid tumours

I M Frayling. Mol Pathol. 2002 Apr.

Abstract

Most mutation detection techniques are unsuitable for routine use on solid tumours. Important parameters include sensitivity, specificity, efficiency, use of existing resources, and cost. In the UK, < 0.2% of service genetics laboratory activity involves mutation analysis in tumours (usually for family studies), mainly because it is time consuming/labour intensive (thus expensive) and DNA extracted from formalin fixed, paraffin wax embedded tissue is of low quality and yield. The small size of DNA fragments obtained from tissue blocks limits the polymerase chain reaction, the basis of most mutation detection methods. Other, biological, factors include: (1) heterogeneity of mutations within and between tumours, (2) variation in type and site of mutations in any one gene, (3) normal tissue harbouring mutations, (4) few genes are mutated in most of any one tumour type, and (5) few clinically useful correlations with genetic changes have been found. Present research is centred on correlating single gene mutations with various clinicopathological features, but the pattern of mutations in a combination of genes will probably prove more useful. Microsatellite instability, however, appears to be worth testing for in both familial and sporadic tumours, particularly of the colorectum.

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Figures

Figure 1
Figure 1
Genetic anatomy.
Figure 2
Figure 2
PCR: the polymerase chain reaction.
Figure 3
Figure 3
Low power (A and B) and high power (C and D) views of sections of a colorectal adenocarcinoma, stained immunohistochemically for the mismatch repair (MMR) proteins MSH2 (A and C) and MLHI (B and D) (brown) with a toluidine blue counterstain. Nuclear staining for MSH2 is preserved in this cancer, whereas the absence of staining for the MLH I protein shows that its expression has been lost. This is consistent with the tumour having arisen in an individual from a family affected by hereditary non-polyposis colorectal cancer and known to carry a germline mutation in the MLHI gene. In the low power views the lymphoid follicles provide a convenient internal control, as do the normal stromal cells and lymphocytes in the high power view. (Figure provided by M Arends, Department of Pathology, University of Cambridge, UK.)
Figure 4
Figure 4
Suggested mismatch repair (MMR) testing strategy in families suspected of hereditary non-polyposis colorectal cancer (HNPCC). IHC, immunohistochemistry; MSI, microsatellite instability.
Figure 5
Figure 5
Suggested mismatch repair (MMR) testing strategy for sporadic colorectal cancers. The percentages are estimates of all cancers presenting, as determined by modelling. HNPCC, hereditary non-polyposis colorectal cancer; MSI, microsatellite instability.
See this image and copyright information in PMC

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