MLH1 mediates PARP-dependent cell death in response to the methylating agent N-methyl-N-nitrosourea.

ABSTRACT:

Background

Methylating agents such as N-methyl-N-nitrosourea (MNU) can cause cell cycle arrest and death either via caspase-dependent apoptosis or via a poly(ADP-ribose) polymerase (PARP)-dependent form of apoptosis. We wished to investigate the possible role of MLH1 in signalling cell death through PARP.

Methods

Fibroblasts are particularly dependent on a PARP-mediated cell death response to methylating agents. We used hTERT-immortalised normal human fibroblasts (WT) to generate isogenic MLH1-depleted cells, confirmed by quantitative PCR and western blotting. Drug resistance was measured by clonogenic and cell viability assays and effects on the cell cycle by cell sorting. Damage signalling was additionally investigated using immunostaining.

Results

MLH1-depleted cel

SUBMITTER: McDaid JR 

PROVIDER: S-EPMC2720233 | biostudies-literature | 2009 Aug

REPOSITORIES: biostudies-literature