癌基因pim-3的克隆及其對肝癌細胞凋亡的影響.pdf

1、南昌大學(xué)醫(yī)學(xué)院碩士學(xué)位論文癌基因pim3的克隆及其對肝癌細胞凋亡的影響姓名：鄧歡申請學(xué)位級別：碩士專業(yè)：內(nèi)科學(xué)（消化系?。┲笇?dǎo)教師：張吉翔20070522Abstract—————————————————————————————————————————————————————————一一一_ABSTRACTAIM：Tocloneratp／m一3cDNAandconstructitspim3eukaryoticexpressingplas

2、midandtoinvestigatetheexpressionofpim3ineukaryocyteanditseffectOilapoptosisMETHODS：TotalRNAwasisolatedfromratskeletalmuscletissueswithTRIz01pim3eDNAwasmadebyRTPCRandconstructedintoeukaryoticexpressionvectorpEGFPN2，whiche

3、xpressesgreenfluorescentproteinTherecombinantpEGFPN2|p／m一3WasthentransformedintoEcoliJMl09hostbacteriaAfterauthenticatedbyrestrictiondigestiveenzymesandsequencing，plasmidpEGFPN260im一3WaStransfectedintoSMMC一7721cellsbylip

4、osomeFluorescentmicroscopeWaSusedtoexaminetheexpressionofpim一3，andthebiologicbehavioroftransfectedcellswastestedbyflowcytometryandMTTRESULTS：Restrictionenzymedigestionanalysisandsequencingshowedthattherecombinantplasmido

5、fratpim一3Wascorrectlyconstructed；FlowcytometryshowedthepercentageofapoptosiscellstransfectedbytherecombinantplasmidpEGFP—N2仞im一3Was348％，andtheplasmidpEGFPN2transfectedcellgroupWaS1074％儼如05)，andnon—transfectedcellgroupWas

6、1101％thereweresignicantdifferencesbetweentherecombinantplasmidtransfectedgroupandtheothertwocontrolgroups(氏005)；FluorescentmicroscopeandMTTshowedapim3expressioninSMMC一7721cellandtheapoptosisofthetransfectedcellsWassignif