辐射诱导重组质粒pegr-p16在黑素瘤b16细胞中表达特性与凋亡作用word格式论文.docxVIP

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辐射诱导重组质粒pegr-p16在黑素瘤b16细胞中表达特性与凋亡作用word格式论文.docx

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辐射诱导重组质粒pegr-p16在黑素瘤b16细胞中表达特性与凋亡作用word格式论文

ExpressionPropertiesandApoptosisEffectsofpEgr-P16 RecombinantPlasmidsinMelanomaB16 Cells Induced byIonizing Irradiation.ABSTRACTBACKGROUNDANDAIM：Inthisstudy,accordingtothemechanismthationizing radiationcanactivateearlygrowthresponse-1(Egr-1)genepromoter,whichinducestheexpressionofdownstreamgenesandanti-tumoreffectofatumorsuppressorgenep16,the studyontheexpressionpropertyofpEgr-p16recombinedplasmidinmelanomaB16cells andtheidentificationofitsantitumoureffectswhileitscombinationwithirradiationwereperformed toestablish anapproachforclinical treatment of melanoma.MATERIALSANDMETHODS：Basedontheconstructionofarecombinantplasmid which containing human suppressor gene p16 and radio-sensitive promoter (Egr-1),pEgr-p16plasmidswaspackagedwithliposomeandtransfectedintomelanomaB16cell line.QuantitativeRT-PCRwasemployedtodetecttheexpressionofp16inmelanomaB16 cellsexposedtothedifferentdosesofX-rayirradiationandtoobserveitsexpression time-courseafterexposuretothesamedoseofirradiation.Flowcytometrywasdoneto showthechangesofapoptosisofthemelanomaB16cells.MTTwasperformedtodetectthe anti-tumor effect of pEgr-p16 combined with irradiation.RESULTS：pEgr-p16recombinantplasmidsweresuccessfullytransfectedintomelanoma B16cells.p16expressionsweresignificantlyincreasedinallgroupsexposedtoX-rayirradiationwithdifferentdosesandwereabout3.78-6.67timeshigherthanthatinshameirradiation group (P0.01). p16 expressions were gradually increase with time afterexposureto2GyX-rayirradiationandreachedmaximum72hlater.Apoptosisin melanomaB16cellscouldbeinducedviap16expressioncombinedwithirradiation,inwhichtheratewashigherthanthatineitherplasmidorirradiationgroupalone(P0.05～0.01). Thenumber of pEgr-p16-transfectedB16 cells exposedto 2GyX-rayirradiation was significantlyless than those in other experimentalgroups (P0.05～0.001).CONCLUSIONS：pEgr-p16genecombinedwithirradiationareabletosuppress melanomaB16cellgrowthsignificantlyanditsinhibitioneffectwasbetterthanthatingene-orirradiation-treatmentalone.Ourresultsareofs