辐射诱导重组质粒pEgrP16在黑素瘤B16细胞中的表达特性与凋亡作用

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1、汕头大学医学院硕士学位论文ExpressionPropertiesandApoptosisEffectsofpEgr-P16RecombinantPlasmidsinMelanomaB16CellsInducedbyIonizingIrradiation.ABSTRACTBACKGROUNDANDAIM：Inthisstudy,accordingtothemechanismthationizingradiationcanactivateearlygrowthresponse-1(Egr-1)genepromot

2、er,whichinducestheexpressionofdownstreamgenesandanti-tumoreffectofatumorsuppressorgenep16,thestudyontheexpressionpropertyofpEgr-p16recombinedplasmidinmelanomaB16cellsandtheidentificationofitsantitumoureffectswhileitscombinationwithirradiationwereperformedtoe

3、stablishanapproachforclinicaltreatmentofmelanoma.MATERIALSANDMETHODS：Basedontheconstructionofarecombinantplasmidwhichcontaininghumansuppressorgenep16andradio-sensitivepromoter(Egr-1),pEgr-p16plasmidswaspackagedwithliposomeandtransfectedintomelanomaB16celllin

4、e.QuantitativeRT-PCRwasemployedtodetecttheexpressionofp16inmelanomaB16cellsexposedtothedifferentdosesofX-rayirradiationandtoobserveitsexpressiontime-courseafterexposuretothesamedoseofirradiation.FlowcytometrywasdonetoshowthechangesofapoptosisofthemelanomaB16

5、cells.MTTwasperformedtodetecttheanti-tumoreffectofpEgr-p16combinedwithirradiation.RESULTS：pEgr-p16recombinantplasmidsweresuccessfullytransfectedintomelanomaB16cells.p16expressionsweresignificantlyincreasedinallgroupsexposedtoX-rayirradiationwithdifferentdose

6、sandwereabout3.78-6.67timeshigherthanthatinshame4汕头大学医学院硕士学位论文irradiationgroup(P<0.01).p16expressionsweregraduallyincreasewithtimeafterexposureto2GyX-rayirradiationandreachedmaximum72hlater.ApoptosisinmelanomaB16cellscouldbeinducedviap16expressioncombinedwit

7、hirradiation,inwhichtheratewashigherthanthatineitherplasmidorirradiationgroupalone(P<0.05～0.1).ThenumberofpEgr-p16-transfectedB16cellsexposedto2GyX-rayirradiationwassignificantlylessthanthoseinotherexperimentalgroups(P<0.05～0.001).CONCLUSIONS：pEgr-p16genecom

8、binedwithirradiationareabletosuppressmelanomaB16cellgrowthsignificantlyanditsinhibitioneffectwasbetterthanthatingene-orirradiation-treatmentalone.Ourresultsareofsubstantialimportantceininvesting