抑制ezh2基因表达对t细胞淋巴瘤放射敏感性的影响

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1、天津医科大学硕士学位论文AbstractObjective:Toinvestigatetheanti-tumorfunctionofEZH2down-regulationcombinedwithirradiationonTcellnon-Hodgkin'slymphoma(T-NHL)invitro,andelucidatetheeffectsoncellproliferationaswellasapoptosisinT-NHLcelllinesHut78andJurkat.Demonstratingifinhibit

2、ionofEZH2couldenhancetheradiosensitivityofT-NHL,wetrytoprovidenewideasandtheoreticalbasisforthegenetherapyincombinationwithconventionalradiotherapy.Methods:1.TheT-NHLcelllinesHut78andJurkatwereinfectedbylentivirus-deliveredEZH2-shRNAandnegativecontrol(emptyvecto

3、r)respectively.2.TheEZH2expressioninmRNAandproteinlevelsofHut78andJurkatweredetectedbyRT-qPCRandWestern-blot.3.MTS/PMScellproliferationassaywasconductedtoevaluatetheeffectofEZH2-shRNAoncellproliferationandradiosensitivitytoXrayinHut78andJurkatcells.4.EZH2inhibit

4、or(UNC1999)wasusedtotreatHut78andJurkatcells,andcausedpharmacologicalinhibitionofEZH2confirmedbyRT-qPCRandWestern-blot.5.MTS/PMScellproliferationassaywasconductedtoevaluatetheeffectofEZH2inhibitor(UNC1999)oncellproliferationandradiosensitivitytoXrayinHut78andJur

5、katcells.6.FlowcytometrywasusedtotoevaluateapoptosisofEZH2inhibitor(UNC1999)incombinationwithirradiationinHut78andJurkatcells7.Rhodamine123stainingwasusedtoobservemitochondriamembranepotentialofEZH2inhibitor(UNC1999)incombinationwithirradiationinHut78andJurkatce

6、lls8.ProteinlevelofBcl-2andcleavedPARPweretestedbyWestern-blotinHut78andJurkatcellstreatedwithEZH2inhibitor(UNC1999)andirradiation.Results：III万方数据天津医科大学硕士学位论文1.Hut78andJurkatcellsweresuccessfullyinfectedbylentivirus-deliveredEZH2-shRNAandemptyvirus,andwegottwotr

7、ansfectedcelllinesofHut78andJurkatrespectively.RT-qPCRandWestern-blotwereperformedtodetecttheexpressionlevelofEZH2.EZH2mRNAlevelsofHut78andJurkatcellsinfectedwithEZH2-shRNAwereonly31.6%and46.7%,lowerthanothertwogroups(P<0.05),whilenosignificantdifferencewasobser

8、vedbetweenuntreatedcellsandnegativecontrol.Western-blotshowedthatexpressionofEZH2proteininHut78EZH2-shRNAandJurkatEZH2-shRNAcellsweresignificantlylowerthanothertwogro