siRNA抑制bcr-abl融合基因的表達(dá)及其對K562細(xì)胞增殖和分化的影響.pdf

1、內(nèi)蒙古醫(yī)學(xué)院碩士學(xué)位論文siRNA抑制bcrabl融合基因的表達(dá)及其對K562細(xì)胞增殖和分化的影響姓名：劉洋申請學(xué)位級別：碩士專業(yè)：內(nèi)科學(xué)指導(dǎo)教師：石玉濤馬玉珍20100501。內(nèi)蒙古醫(yī)學(xué)院碩士研究生學(xué)位論文(2010年)Expressionoffusiongenebcr‘‘a(chǎn)blsuppressionbysiRNAanditsroleinproliferationanddifferentiationofK562cellsAbstrac

2、tObjectiveTooonstructRNAivectortargetingfusiongenebcr?！痑blandinvestigateitsmlesintheproliferationanddifferentiationofK562cellsaftertransfeetionMethodsThrL孢sequence—specificsiRNAsvectorsbasedonthegenesequence0ffusiongeneb

3、er—abl，negtivecontrolwithouthomologoussequenceandpositivevectorcontroltargetingGAPDHwereconstructedandcardedneomycinresistanceandGFPandIU甲forselectionfortransgeniccellsandtOdeterminetransfectioneffidency：(1)VectorDNAwast

4、ransfectedintochronicmyeloidleukemiacelllineK562usingLiposome—mediatedmethod，andpositivecloneswereselectedbyneomycinG418andfluorescence；(2)rnRNAexpressionlevelSoffusiongenebdablinK562cellsaftertransfectionweredetectedbyR

5、T—PCR；(3)T11econditionsofdifferentiationfromtransgeniccellstoerythroidcellsbysiRNAinterferencewasobservedusingBenzidinestaining；(4)TransgeniccellsselectedbyRNAinterferenceafter15dayswereusedtOdetectitssensitivitytoAra—c，

6、Interferon—aandcytosinearabinosidecombineda—interferonbyTrypanbluestainingResults(1)Usinglaseroonfocalfluorescencemicroscope，thepercentofK562cellsaccountfor40％byLiposome—mediatedtransfectionafter48handthepercentreached95

7、％withG418selectionafter15days；(2)ComparedtOcon—trois，mRNAexpressionlevelsofthethreetargetsitesonfusiongenebcr—ablWassignificandylower，especiallyonsitetwo；(3)48haftersiRNAinterference，differentiatedcellsfromtrmlsgeniccell

8、stOerythroidcellsontargetone，twoandthreeincreasedfrom(380040)％to(1820262)％、(3333245)％、(1653114)％incontrasttoeontrols，respectively；15dayslater，thepercentageoftransgentieceilswereincreasedform(380060)％to(5307130％、(61872122