黑素瘤A375細(xì)胞系中cFLIPp43促進(jìn)NF-κB信號(hào)通路的活化.pdf

1、華中科技大學(xué)碩士學(xué)位論文黑素瘤A375細(xì)胞系中cFLIPp43促進(jìn)NFκB信號(hào)通路的活化姓名：陸捷潔申請(qǐng)學(xué)位級(jí)別：碩士專業(yè)：皮膚病與性病學(xué)指導(dǎo)教師：涂亞庭20090501華中科技大學(xué)碩士學(xué)位論文華中科技大學(xué)碩士學(xué)位論文cFLIPp43inducesactivationoftheNFκBsignalingpathwayinmelanomaA375celllineMasterCidate:LuJiejieSupervis:Prof.TuYa

2、tingAbstractObjectiveOurstudyexaminedtheexpressionofcFLIPNFκBp65inhumanmelanomaA375SKMel1celllineonproteinleveltoinvestigatewhethertransfectionofA375celllinewithpCMVTag2BcFLIPp43expressionvectcaninducetheactivationofNFκB

3、signalingpathwaynot.MethodsWeinvestigatedtheproteinexpressionlevelsofcFLIPNFκBp65inhumanmelanomaA375SKMel1cellline.WefurtherconstructedaeukaryoticexpressionvectfcFLIPp43withtheplaspCMVTag2B.ThemonocloneA375cellswithstabl

4、eexpressionofcFLIPp43wereobtainedbyG418ionwereidentifiedwithwesternblot.WeinvestigatedtheproteinexpressionlevelofNFκBp65inA375celllinewithstableexpressionofcFLIPp43.Theimmunofluescencetechnologywasusedtoexaminethenuclear

5、translocationofNFκBp65instabletransfectedA375cellline.ThenwetransfecteddifferentdosesofpCMVTag2BcFLIPp43expressionvectintoA375celllinetheactivationofNFκBsignalingpathwaywasvaluedbytheDualLuciferaseRepterAssaySystem.Resul

6、tsTheproteinexpressionlevelofcFLIPNFκBp65inA375celllinewaslowerthaninSKMel1cellline.WesuccessfullyconstructedA375celllinewhichstablyexpressedcFLIPp43.TheproteinexpressionlevelofNFκBp65washigherinA375celllinewithstableexp

7、ressionofcFLIPp43thaninA375cellline.WiththeenhancementofcFLIPp43proteinexpressiontheamountofnuclearlocationofNFκBp65increased.TheNFκBactivationaugmentedwiththeincreaseofpCMVTag2BcFLIPp43instanttransfectioninA375cellline.