A Cell-Free Assay for the Functional Analysis of Variants of the Mismatch Repair Protein MLH1

作者：Drost Mark; Zonneveld Jose B M; van Dijk Linda; Morreau Hans; Tops Carli M; Vasen Hans F A; Wijnen Juul T; de Wind Niels^*

来源：Human Mutation, 2010, 31(3): 247-253.

DOI：10.1002/humu.21180

• Lynch syndrome
• DNA mismatch repair
• MMR
• variants of uncertain significance
• functional assay
• MLH1

The hereditary colon and endometrium cancer predisposition Lynch Syndrome (also called HNPCC) is caused by a germ-line mutation in one of the DNA mismatch repair (MMR) genes. A significant fraction of the gene alterations detected in suspected Lynch Syndrome patients is comprised of amino acid substitutions. The relevance for cancer risk of these variants is difficult to assess, as currently no time- and cost-effective, validated, and widely applicable functional assays for the measurement of MMR activity are available. Here we describe a rapid, cell-free, and easily quantifiable MMR activity assay for the diagnostic assessment of variants of the MLH1 MMR protein. This assay allows the parallel generation and functional analysis of a series of variants of the MLH1 protein in vitro using readily available, or preprepared, reagents. Using this assay we have tested 26 MLH1 variants and of these, 15 had lost activity. These results are in concordance with those obtained from first-generation assays and with in silico and pathology data. After its multifocal technical and clinical validation this assay could have great impact for the diagnosis and counseling of carriers of an MLH1 variant and their relatives. Hum Mutat 31:247-253, 2010.

• 出版日期2010-3