Detection of DNA mismatch repair proteins in fresh human blood lymphocytes - towards a novel method for hereditary non-polyposis colorectal cancer (Lynch syndrome) screening

Journal of Experimental & Clinical Cancer Research

Jeremy Z Fields¹  Akhtar A Ali³  Zohra J Ali-Khan Catts²  Chandra Somerman²  Marcie Parker²  Nawab Ali³  Bruce M Boman²  Samar Hassen³ 

[1] CATX Inc., Gladwyne PA 19035, USA;Helen F Graham Cancer Center, Newark DE 19713, USA;Graduate Institute of Technology, University of Arkansas at Little Rock, Little Rock, AR 72204, USA

关键词: Cell Culture;    Western blotting;    PHA treatment;    Lymphocytes;    MSH2;    MLH1;    HNPCC;    MMR proteins;    Hereditary Cancer;    Lynch Syndrome;   

Others  :  827131
DOI  :  10.1186/1756-9966-30-100

 received in 2011-05-30, accepted in 2011-10-21,  发布年份 2011

【 摘 要 】

Background

A broad population-based assay to detect individuals with Lynch Syndrome (LS) before they develop cancer would save lives and healthcare dollars via cancer prevention. LS is caused by a germline mutation in a DNA mismatch repair (MMR) gene, especially protein truncation-causing mutations involving MSH2 or MLH1. We showed that immortalized lymphocytes from LS patients have reduced levels of full-length MLH1 or MSH2 proteins. Thus, it may be feasible to identify LS patients in a broad population-based assay by detecting reduced levels of MMR proteins in lymphocytes.

Methods

Accordingly, we determined whether MSH2 and MLH1 proteins can also be detected in fresh lymphocytes. A quantitative western blot assay was developed using two commercially available monoclonal antibodies that we showed are specific for detecting full-length MLH1 or MSH2. To directly determine the ratio of the levels of these MMR proteins, we used both antibodies in a multiplex-type western blot.

Results

MLH1 and MSH2 levels were often not detectable in fresh lymphocytes, but were readily detectable if fresh lymphocytes were first stimulated with PHA. In fresh lymphocytes from normal controls, the MMR ratio was ~1.0. In fresh lymphocytes from patients (N > 50) at elevated risk for LS, there was a bimodal distribution of MMR ratios (range: 0.3-1.0).

Conclusions

Finding that MMR protein levels can be measured in fresh lymphocytes, and given that cells with heterozygote MMR mutations have reduced levels of full-length MMR proteins, suggests that our immunoassay could be advanced to a quantitative test for screening populations at high risk for LS.

【 授权许可】

   
2011 Hassen et al; licensee BioMed Central Ltd.

【 图 表 】

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