title: Chk1 is essential for tumor cell viability following activation of the replication checkpoint.
abstract: Chk1 (checkpoint kinase 1) is an evolutionarily conserved serine/threonine kinase involved in DNA damage responses. Originally identified as a kinase regulating the G2/M transition checkpoint, its role has broadened to include the S-phase checkpoint response and essential functions in early embryonic development. In this manuscript we investigated the potential of chemo-sensitization via ablation of Chk1 in cells treated with anti-metabolite cancer drugs, hydroxyurea (HU) and cytosine arabinoside (ara-C). Exposure to these replication interfering drugs in cells carrying Chk1 targeted siRNA provoked markedly increased rates of apoptosis. Although cell death was accompanied by an increase in p53 and activation of Chk2, the increased susceptibility to apoptosis was not dependent on p53 or Chk2. Additionally, we found that cells with reduced Chk1 expression displayed increased gamma-H2A.X expression, a marker for damaged DNA, and phosphorylated 32kDa subunit of replication protein A (RPA). Thus, Chk1 may play an essential role in maintaining DNA integrity during the replication block. Significantly, normal cells such as WS1 did not exhibit increased DNA damage or subsequent increases in apoptosis following replication stress, in the absence of Chk1. Thus, the essential role Chk1 plays in maintaining viability during the replication block in cancer cell lines can be exploited to sensitize cancer cells when abrogation of Chk1 is combined with DNA anti-metabolite chemotherapeutic drugs. Taken together, these data suggest that inhibition of Chk1 in combination with DNA anti-metabolite chemotherapy is a viable therapeutic strategy.
authors.affiliation: Department of Discovery Research, DNAX Research Incorporated, Palo Alto, California, USA.
authors.forename: Song H
authors.initials: SH
authors.lastname: Cho
authors.forename: Christian D
authors.initials: CD
authors.lastname: Toouli
authors.forename: Gregory H
authors.initials: GH
authors.lastname: Fujii
authors.forename: Chad
authors.initials: C
authors.lastname: Crain
authors.forename: David
authors.initials: D
authors.lastname: Parry
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000235:genetics / Q000502:physiology; D002453:Cell Cycle / Q000235:genetics / Q000502:physiology; D045744:Cell Line, Tumor; D002470:Cell Survival; D000071877:Checkpoint Kinase 1; D064447:Checkpoint Kinase 2; D003561:Cytarabine / Q000494:pharmacology; D004249:DNA Damage / Q000235:genetics / Q000502:physiology; D004261:DNA Replication / Q000187:drug effects; D004273:DNA, Neoplasm / Q000235:genetics; D006657:Histones / Q000235:genetics / Q000502:physiology; D006801:Humans; D006918:Hydroxyurea / Q000494:pharmacology; D011494:Protein Kinases / Q000235:genetics / Q000502:physiology*; D017346:Protein Serine-Threonine Kinases / Q000235:genetics / Q000502:physiology; D034741:RNA, Small Interfering / Q000187:drug effects / Q000502:physiology; D051716:Replication Protein A / Q000235:genetics / Q000502:physiology; D016159:Tumor Suppressor Protein p53 / Q000235:genetics / Q000502:physiology
chemical_list: D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; C468783:H2AX protein, human; D006657:Histones; D034741:RNA, Small Interfering; D051716:Replication Protein A; D016159:Tumor Suppressor Protein p53; D003561:Cytarabine; D011494:Protein Kinases; D064447:Checkpoint Kinase 2; C000605867:CHEK1 protein, human; C578634:CHEK2 protein, human; D000071877:Checkpoint Kinase 1; D017346:Protein Serine-Threonine Kinases; D006918:Hydroxyurea