MSRE-PCR for analysis of gene-specific DNA methylation.

Abnormal DNA methylation is observed in certain promoters of neoplastic cells, although the likelihood of methylation for each individual promoter varies. Simultaneous analysis of many promoters in the same sample can allow use of statistical methods for identification of neoplasia. Here we describe an assay for such analysis, based on digestion of genomic DNA with methylation-sensitive restriction enzyme and multiplexed PCR with gene-specific primers (MSRE-PCR).

Overexpression of DcR1 and survivin in genetically modified cells with pleiotropic drug resistance.

A previously identified set of short cDNA fragments (genetic suppressor elements, GSEs) expressed in human HT1080 cells protects them against several chemotherapeutic drugs. We show here that DNA damaging agent cytosine arabinoside can induce apoptosis in GSE-containing drug-resistant derivatives (M125 cells) of HT1080, suggesting that apoptotic pathways are preserved in M125. We also show that both parental cells and M125 constitutively express Fas ligand and TNF-related apoptosis inducing ligand, thus pre-disposing cells to apoptosis.