Molecular Mechanisms of Paraptosis Induction Implications for a Non-Genetically Modified Tumor Vaccine 英文参考文献.docVIP

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Molecular Mechanisms of Paraptosis Induction Implications for a Non-Genetically Modified Tumor Vaccine 英文参考文献.doc

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Molecular Mechanisms of Paraptosis Induction Implications for a Non-Genetically Modified Tumor Vaccine 英文参考文献

MolecularMechanismsofParaptosisInduction: ImplicationsforaNon-GeneticallyModifiedTumor Vaccine NeilHoa1,MichaelP.Myers3,ThomasG.Douglass5,JianGangZhang1,2,ChristinaDelgado1 ,Lara Driggers1,LindaL.Callahan4,GeraldVanDeusen3,JimmyT.H.Pham3,NiravBhakta3,LishengGe1, MartinR.Jadus1,2,6 * 1DiagnosticandMolecularMedicineHealthcareGroup,VeteransAffairsMedicalCenter,LongBeach,California,UnitedStatesofAmerica,2DepartmentofPathologyand LaboratoryMedicine,UniversityofCaliforniaIrvine,Irvine,California,UnitedStatesofAmerica,3ChemistryandBiochemistryDepartment,CaliforniaStateUniversityLong Beach,LongBeach,California,UnitedStatesofAmerica,4NursingDepartment,CaliforniaStateUniversityLongBeach,LongBeach,California,UnitedStatesofAmerica, 5BiologyDepartment,CaliforniaStateUniversityLongBeach,LongBeach,California,UnitedStatesofAmerica,6Neuro-OncologyProgram,ChaoComprehensiveCancer Center,UniversityofCaliforniaIrvine,Irvine,California,UnitedStatesofAmerica Abstract Paraptosisistheprogrammedcelldeathpathwaythatleadstocellularnecrosis.Previously,rodentandhumanmonocytes/ macrophages killed glioma cells bearing the membrane macrophage colony stimulating factor (mM-CSF) through paraptosis,butthemolecularmechanismofthiskillingprocesswasneveridentified.Wehavedemonstratedthatparaptosis of rat T9 glioma cells can be initiated through a large potassium channel (BK)-dependent process initiated by reactive oxygen species. Macrophage mediated cytotoxicity upon the mM-CSF expressing T9-C2 cells was not prevented by the addition of the caspase inhibitor, zVAD-fmk. By a combination of fluorescent confocal and electron microscopy, flow cytometry, electrophysiology, pharmacology, and genetic knock-down approaches, we demonstrated that these ion channelscontrolcellularswellingandvacuolizationofratT9gliomacells.Celllysisisprecededbyadepletionofintracellular ATP.Six-hourexposuretoBKchannelactivationcausedT9cellstooverexpressheatshockproteins(Hsp60,70,90and gp96).ThissametreatmentforcedHMGB1translocationfr