吉马酮抑制肝癌bel7402细胞增殖机制初步分析-preliminary analysis on the mechanism of gemma ketone inhibiting proliferation of hepatocellular carcinoma bel 7402 cells.docxVIP

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吉马酮抑制肝癌bel7402细胞增殖机制初步分析-preliminary analysis on the mechanism of gemma ketone inhibiting proliferation of hepatocellular carcinoma bel 7402 cells.docx

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吉马酮抑制肝癌bel7402细胞增殖机制初步分析-preliminary analysis on the mechanism of gemma ketone inhibiting proliferation of hepatocellular carcinoma bel 7402 cells

AbstractRhizomacurcuma,avaluableChinesetraditionalmedicine,hasbeenmassivelyappliedintoclinicaltherapyinChinesemedicinebecauseofitsanti-inflammatory,hepatoprotective,anti-bacterial,anti-thromboticandanti-tumorproperties.GermacroneisakindofsesquiterpeneextractedfromR.curcuma,andhasbeenreportedtopossessplentyofpharmacologicalpropertieswhiletheanti-cancermechanismhasnotthoroughlyunderstoodyet.ThepresentstudyistoclarifytheeffectofgermacroneonBel7402throughMTT,FlowCytometry,WesternBlottingandsoon.Andthemainresearchresultsareasfollows.（1）GermacroneselectivelyinhibitedtheproliferationofBel7402cellsindose-dependentmannerwhileexhibitednoobvioustime-dependenttendency.TheMTTassayresultsshowedhighergermacroneconcentrationcausedhighergrowthinhibitoryrateofBel7402cells.Andtherewasnostatisticallysignificantdifferencebetweentreatmentsfor24hand48h.IC50for24htreatmentwithgermacronewas160μM.Furthermore,thesametreatmentsonnormalhumanlivercellLO2cellsrevealedlittleside-effects.（2）Germacronedose-dependentlyinducedtheapoptosisofBel7402cellsandraiseditsoxidativedamage.BymeansoftheHocheststainingandFlowcytometryassay,wefoundthathigherconcentrationofdrugledtothemoreobviousinhibitoryeffect.Inaddition,theWesternBlottingresultsdemonstratedthatgermacroneinducedtheexpressionofanti-cancergeneP53andBaxwhilerepressedtheexpressionofanti-apoptosisgeneBcl-2.Furthermore,germacroneefficientlyup-regulatedtheROSlevelinBel7402cells.AlltheresultsexhibitedthatgermacronehadthepotentialtopromotetheapoptosisandinvolvedintheoxidativedamageinBel7402cells.（3）GermacroneinducedtheapoptosisofBel7402cellsthroughtheJAK2/STAT3signalpathway.WesetseveraldifferenttreatmentsonBel7402cellsbyusinggermacroneandIL-6,theagonistofJAK2/STAT3signalpathway,andtheWesternBlottingresultsindicatedthatgermacronedown-regulatedtheexpressionlevelofp-STAT3andIL-6hadtheoppositeeffect,inthemeanwhile,theexpressionlevelofP53showedoppositetrend.Inconclusion,alltheresultsdemonstratedthatgermacronesuppressedthephosphorylationofconstitutionalactivatio