Inactivating mutations of the caspase-10 gene in gastric cancer Won Sang Park 1,3 , Jong Heun Lee 1,3 , Min Sun Shin 1 , Jik Young Park 1 , Hong Sug Kim 1 , Jong Ho Lee 1 , Young Sil Kim 1 , Shi Nae Lee 1 , Wenhua Xiao 1 , Cho Hyun Park 2 , Sug Hyung Lee 1 , Nam Jin Yoo 1 and Jung Young Lee* ,1 1 Department of Pathology, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea; 2 Department of Surgery, College of Medicine, The Catholic University of Korea, Seoul 137-701, Korea We have analysed the genetic alteration of the entire coding region and all splice sites of caspase-8 and -10 genes in 99 gastric cancers by polymerase chain reaction (PCR)-single strand conformation polymorphism (SSCP) and sequencing. We found LOH of the caspase-8 and -10 in nine (28%) of 32 and in four (15%) of 26 informative cases, respectively. Overall, three of 99 gastric cancers (3%) were found to have the caspase-10 mutations, which were identi®ed in the coding regions of the death eector domain (codon 147) and the p17 large protease domain (codons 257 and 410), whereas no mutation was detected in caspase-8. In vitro expression studies, the M147T and Q257stop mutants severely impaired caspase-10-mediated apoptosis, whereas the V410I which was the same mutation detected in ALPS patient had a signi®cant, albeit less severe, eect on apoptosis. The data presented here suggest that somatic alterations of the caspase-10 gene might contribute to the pathogenesis in a subset of gastric cancers through the loss of their apoptotic function. Oncogene (2002) 21, 2919 ± 2925. DOI: 10.1038/sj/ onc/1205394 Keywords: caspase-10; mutation; LOH; gastric cancer; apoptosis Introduction Accumulation of abnormal numbers of a clonally related population (neoplasm) can result from either uncontrolled proliferation or inhibition of cell death. The death receptor-ligand systems such as Fas-Fas receptor, TNF-TNF receptor, and TRAIL-TRAIL receptor are important pathways for the negative growth regulation of cells and tissues. Binding of death receptor by its agonistic antibodies or by its ligand induces the self-association of a conserved cytoplasmic region of death receptor called the death domain. This death receptor homotrimer interacts with a protein called Fas-associated death domain (FADD) containing another death domain and recruits the inactive procaspase-8 and/or -10 by FADD molecule and forms death-induced signaling complex (DISC). Aggregation of procaspase-8 and/or -10 in DISC initiates autoprocessing and release of active form of caspase-8 and -10 into the cytoplasm, where a proteolytic cascade leads to apoptosis (Fernandes- Alnemri et al., 1996; Ng et al., 1999; Vincenz and Dixit, 1997; Wang et al., 1999). Disruption of this process may result in illegitimate cell survival and contribute to several human diseases including auto- immune lymphoproliferative syndrome (ALPS) or neoplasia (Nagata, 1997; Thompson, 1995). ALPS (autoimmune lymphoproliferative syndrome) is an inherited disease associated with germline mutation of either Fas (ALPS type Ia) or its ligand FasL (ALPS type Ib), which result in marked over- accumulation of mature lymphocytes and profound lymphadenopathy due to failure of Fas-mediated apoptosis. Recently Wang et al. (1999) reported germline mutation of caspase-10 gene in two kindreds with ALPS type II, but not of other death-related genes such as Fas, FasL, TNF receptor-1, FADD, TRADD, RIP, caspases-7, -8 or -9. Thus, they concluded that caspase-10 was the gene responsible for the ALPS type II phenotype in these pedigrees. Gastric cancer occurs with a high incidence in Asia including Korea and is one of the leading causes of cancer death in the world. However, the molecular pathogenesis of gastric cancer is still unclear. There are accumulating evidences that human cancers including gastric cancer cells have escape mechanisms from death receptor-mediated apoptosis such as the mutations of primary structure of death receptors (Grùnbñk et al., 1998; Fisher et al., 1995; Landowsky et al., 1997; Lee et al., 1999; Shin et al., 1999; Park et al., 2001), the production of soluble Fas (Lee et al., 1998b), and ampli®cation of a decoy receptor (Pitti et al., 1998). In addition, chromsome 2q33, where the caspases-8 and -10 genes reside (Fernandes-Alnemri et al., 1996), has been reported as a commonly deleted region in gastric cancer (Nishizuka et al., 1998) and lung cancer (Otsuka et al., 1996). It has also been reported that caspase-8 is frequently inactivated through DNA methylation as well as mutation in neuroblastoma cell lines (Takita et al., 2001). Because caspases-8 and -10 are important Oncogene (2002) 21, 2919 ± 2925 ã 2002 Nature Publishing Group All rights reserved 0950 ± 9232/02 $25.00 www.nature.com/onc *Correspondence: JY Lee; E-mail: stingray@cmc.cuk.ac.kr 3 Won Sang Park and Jong Heun Lee contributed equally to this work Received 29 May 2001; revised 18 December 2001; accepted 7 February 2002 ONCOGENOMICS