49963.原癌基因cmycrna干擾對細胞周期的影響

1、Y774618分類號：密級：’jcI目差≮犬等碩士學位論文單位代碼：10019學號：S02299原癌基因cmycRNA干擾對細胞周期的影響TheeffectofRNAinterferenceagainstcmyconcogeneoncellcycle研究生：途豎定指導教師：造蕉蕉熬援申請學位門類級別：壅堂亟專業(yè)名稱：基趟璺醫(yī)堂研究方向：獸醫(yī)藥堡生重堡堂所在學院：動物匡堂隧2005年6月AbstractC—myeisaeternalonc

2、ogeneandplaysacriticalroleinregulatingcellproliferation，growth，apoptosisanddifferentiationOverexpressionore—mycisoneofthemostcommonalterationsinhumancancersandismostlyresponsibleforthemaglinanttransformationInourstudy，th

3、epSilencerc—myeplasmidexpressingSiRNAagainstcmycmRNAwastransfeetedintoA549cellsbypositiveionmethod72haftertransfection，cellswereharvestedandthelevelsofcmycmRNAweredetectedRTPCRresultsshowedthatc—mycmRNAoftransfectedcells

4、wasdecreasedto40％comparedwiththeuntransfectedcellsAtthesametime，thecMycproteinlevelswereestimatedbyWesternBlottinganddisplayedadecreaseof70％SoitcanbeconcludedthattheRNAinterferenceagainstcmycwassuccessfuJTheFlowcytometry

5、wasemployedtoanalyzetheeffectofcmycRNAionthecellcycleofA549cells72haftertransfectionOurresultsshowedthattheRNAinterferencedecreasedtheproportionofG0／G1phasecellsby16％，from515士17％to351士11％，andthepercentageofSphasecellswas

6、increasedby14％from41，341。4％tO55】士l，6％ButnochangeofG2／Mcelnumberswasobservedbetweencontrolcellsandtransfectedcells(p005)ThesedatadeclaredthatcmyeRNAiinducedaSarrestofcellcycleToinvestigatehowtheSarresthappened，analysesoft

7、hemRNAlevelsofcellcyclerelativegenewereperformed，includingcyclin，CDKandCKIOurexperimentsrevealedthatafterRNAinterference：1)ThemRNAlevelsofcylcinD3，E，Aweremarkedlydownregulatedindifferentextents(p005)2)CDK2andCDK4mRNAleve

8、lsweredecreased(p(o05)，by44士4％，39土4％respectively3)Inoppositiontotheformers，p21andp27mRNAwereelevated(p005)，by366％and44士8％(pO05)Tobeconsciousofwhether“thechangesofmRNAinducedalterationsofproteinexpressionproteinlevelsofCy

9、clinandCDKweredetectedOurdatashowedthat72haftertransfection，theproteinlevelsofcyclinD3，EAandCDK24weredecreased(po05)，by425％、4t2％、575％and569％、365％respectivelyThisconfirmedthatthedownregulationsofcyclinD3，E，AandCDK2，4mRNAw

10、ereresponsiblefordecreasesoftheirproteinAlltheseresultsexplainedthatafterc—myeRNAinterferenceandthedecreasedexpressionoftMyc，theactivationsoncyclinsandCDKsaswellastherepressionsonCKIswereimpairedThisdirectlyinducedtheSar