both hmutsα and hmuts？ dna mismatch repair complexes participate in 5-fluorouracil cytotoxicityhmutsα和hmuts dna错配修复复合物参与5 -氟尿嘧啶细胞毒性.pdfVIP

Both hMutSa and hMutSß DNA Mismatch Repair Complexes Participate in 5-Fluorouracil Cytotoxicity 2. 1,2. 1 2 Akihiro Tajima , Moriya Iwaizumi , Stephanie Tseng-Rogenski , Betty L. Cabrera , John M. Carethers1,2,3* 1 Division of Gastroenterology, Department of Internal Medicine, University of Michigan, Ann Arbor, Michigan, United States of America, 2 Division of Gastroenterology, Department of Medicine, University of California San Diego, La Jolla, California, United States of America, 3 Moores Comprehensive Cancer Center, University of California San Diego, La Jolla, California, United States of America Abstract Background: Patients with advanced microsatellite unstable colorectal cancers do not show a survival benefit from 5- fluorouracil (5-FU)-based chemotherapy. We and others have shown that the DNA mismatch repair (MMR) complex hMutSa binds 5-FU incorporated into DNA. Although hMutSß is known to interact with interstrand crosslinks (ICLs) induced by drugs such as cisplatin and psoralen, it has not been demonstrated to interact with 5-FU incorporated into DNA. Our aim was to examine if hMutSß plays a role in 5-FU recognition. Methods: We compared the normalized growth of 5-FU treated cells containing either or both mismatch repair complexes using MTT and clonogenic assays. We utilized oligonucleotides containing 5-FU and purified baculovirus-synthesized hMutSa and hMutSß in electromobility shift assays (EMSA) and further analyzed binding using surface plasmon resonance. Results: MTT and clonogenic assays after 5-FU treatment demonstrated the most cytotoxicity in cells with both hMutSa and hMutSß, intermediate cytotoxicity in cells with hMutSa alone, and the least cytotoxicity in cells with hMutS