Selective Killing of Cancer Cells by Ashwagandha Leaf Extract and Its Component Withanone Involves ROS Signaling 英文参考文献.docVIP

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Selective Killing of Cancer Cells by Ashwagandha Leaf Extract and Its Component Withanone Involves ROS Signaling 英文参考文献.doc

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SelectiveKillingofCancerCellsbyAshwagandhaLeaf ExtractandItsComponentWithanoneInvolvesROS Signaling NashiWidodo1,2,DidikPriyandoko1,3,NavjotShah1,3,RenuWadhwa1*,SunilC.Kaul1* 1NationalInstituteofAdvancedIndustrialScienceandTechnology(AIST),Tsukuba,Ibaraki,Japan,2DepartmentofBiology,FacultyofMathematicsandNaturalSciences, BrawijayaUniversity,Malang,Indonesia,3GraduateSchoolofLifeandEnvironmentalSciences,UniversityofTsukuba,Tsukuba,Ibaraki,Japan Abstract BackgroundandPurpose:AshwagandhaisapopularAyurvedicherbusedinIndiantraditionalhomemedicine.Ithasbeen assigned a variety of health-promoting effects of which the mechanisms remain unknown. We previously reported the selective killing of cancer cells by leaf extract of Ashwagandha (i-Extract) and its purified component Withanone. In the present study, we investigated its mechanism by loss-of-function screening (abrogation of i-Extract induced cancer cell killing)ofthecellulartargetsandgenepathways. Methodology/PrincipalFindings:Randomizedribozymelibrarywasintroducedintocancercellspriortothetreatmentwith i-Extract.Ribozymeswererecoveredfromcellsthatsurvivedthei-Extracttreatment.Genetargetsoftheselectedribozymes (aspredictedbydatabasesearch)wereanalyzedbybioinformaticsandpathwayanalyses.Thetargetswerevalidatedfor theirroleini-Extractinducedselectivekillingofcancercellsbybiochemicalandmolecularassays.Fifteengene-targetswere identifiedandwereinvestigatedfortheirroleinspecificcancercellkillingactivityofi-Extractanditstwomajorcomponents (WithaferinAandWithanone)byundertakingtheshRNA-mediatedgenesilencingapproach.Bioinformaticsontheselected gene-targetsrevealedtheinvolvementofp53,apoptosisandinsulin/IGFsignalingpathwayslinkedtotheROSsignaling.We examinedtheinvolvementofROS-signalingcomponents(ROSlevels,DNAdamage,mitochondrialstructureandmembrane potential)anddemonstratethattheselectivekillingofcancercellsismediatedbyinductionofoxidativestress. Conclusion:AshwagandhaleafextractandWithanonecauseselectivekillingofcancercellsbyi