Cryosurgery is an emerging treatment for @entity1 solid @entity5 , notably @entity14 . Cryosurgical procedures generate a thermal gradient of from at least -50 degrees C at the center of the @entity5 being treated to about 0 degrees C at the periphery. Cell death occurs by @entity39 in the center, while the peripheral zone of frozen @entity5 harbors a mix of viable and dead tissue. In order to understand the mechanisms of cell death and survival in this peripheral area at risk for @entity5 recurrence, we have established an in vitro freezing system that mimics in vivo conditions of sublethal injury. HT29 @entity694 cells were subjected to freezing temperatures from -6 degrees C to -36 degrees C, thawed at room temperature for 30 min and rewarmed at 37 degrees C for a period of time. Post-freeze-thaw, cryolytic cells were evaluated by @entity3460 exclusive assay. We also identified apoptotic cells after rewarming by cell shrinkage, nucleic condensation, TUNEL assay, DNA fragmentation and @entity12982 degradation. The intensity of cryolysis and apoptosis was increased by lowering the freezing temperature. At -36 degrees C, all cells were dead immediately after freeze-thaw. A kinetic analysis of cryo-induced apoptosis showed that the commitment to enter apoptosis occurred right after the freeze-thaw period and lasted less than 8 hr after rewarming. We further demonstrated that freezing triggers one of the caspase cascade involved in apoptosis: release of @entity4674 from mitochondria to cytosol, followed by activation of @entity8187 and degradation of @entity12982 . These results indicate the death of @entity5 cells under cryo-treatment at sublethal freezing temperature can be attributed 2 different modes, cryolysis as well as apoptosis. HT29 cells carrying @entity876 mutant have very quick response for induction of apoptosis by cryo-treatment and contain an intact pathway of caspase cascade. Further studies will address if mechanisms in cells with wild-type @entity876 will differ.

Induction of apoptosis in @entity1 @entity694 cells HT29 by sublethal cryo-injury: mediation by XXXX release.

@entity1 :: ('9606', 'Species') :: ['human'] @entity39 :: ('MESH:D009336', 'Disease') :: ['necrosis'] @entity5 :: ('MESH:D009369', 'Disease') :: ['tumor', 'cancer', 'tumors'] @entity4674 :: ('54205', 'Gene') :: ['cytochrome c'] @entity12982 :: ('142', 'Gene') :: ['PARP'] @entity8187 :: ('842', 'Gene') :: ['caspase-9'] @entity876 :: ('7157', 'Gene') :: ['p53'] @entity3460 :: ('MESH:D014343', 'Chemical') :: ['trypan blue'] @entity14 :: ('MESH:D015179', 'Disease') :: ['colorectal liver metastasis'] @entity694 :: ('MESH:D003110', 'Disease') :: ['colon cancer']
@entity4674 :: (54205,Gene) :: ['cytochrome c']
