鳥氨酸脫羧酶(ODC)作為抗腫瘤分子靶點的基礎研究.pdf

1、三峽大學碩士學位論文鳥氨酸脫羧酶（ODC）作為抗腫瘤分子靶點的基礎研究姓名：任玉珊申請學位級別：碩士專業(yè)：免疫學指導教師：王艷林韓鈺20100501IIIAbstractObjective:ToclonehumanmouseODCgenetodetermineeffectsofhighexpressionlowexpressionofODContheproliferationchemosensitivityofcancercelllin

2、es.ThepurposeofthisstudywastoexplethepotentialvalueofODCasamoleculartargetfantitumtherapytoprovideanewmeanofcancertreatment.Methods:(1)cDNAcodingfhumannithinedecarboxylase(hODC)wasclonedbyNestRTPCRedintovectpcDNA3.1()toc

3、onstructeukaryoticexpressionplaspcDNA3.1()hODCantisenseplaspcDNA3.1()rODC.PlaspcDNA3.1()rODCwasthentransfectedintohumanhepatomaHepG2cellshumanlymphadenomaJurkatcellstoobtainthecelllineswithstableODCexpressioninhibition.(

4、2)cDNAcodingfmousenithinedecarboxylase(mODC)wasclonedbyNestRTPCRedintovectpcDNA3.1().EukaryoticexpressionplaspcDNA3.1()mODCwasthentransfectedintomousemelanomaB16F1cellstoobtainthecelllinewithODChighexpression.(3)MTTMTSme

5、thodswereusedtoassayproliferationchemosensitivityofcancercells.DNAfragmentationwesternblottingflowcytometryassaywereusedtoobserveapoptosiscellcycle.(4)B16F1celllinewithODChighexpressionwasusedtoinoculateBalbCmousetheneff

6、ectsofODCexpressionlevelontumgrowthwereobserved.Results:(1)HumanmouseODCcDNAweresuccessfullycloned.ThecelllineswithstablelowexpressionhighexpressionofODC(JurkatcelllineJoHepG2celllineHr1B16F1celllineBo)wereobtained.(2)MT

7、TMTSassayshowedthatODClowexpressioncoulddecreasecellgrowthproliferationincreasesensitivitytoantitumdrugs.ApoptoticassaydemonstratedthatthecellswithODClowexpressionweremesensitivetodevelopapoptosisafterantitumdrugtreatmen

8、t.InB16F1cellshighexpressionofODCcouldreducecellsensitivitytoBENSinhibitBENSinducedapoptosis.(3)AnimalexperimentdisplayedthatODChighexpressionraisedthetumgrowthbearingcancerrateofmouse.Conclusion:Inthetumcellsassayedinth

9、isexperimentODChighexpressionstimulatedcellgrowthreducedcellsensitivitytothetestedantitumchemicals.ButODClowexpressioninhibitedcellproliferationmadecellsmesensitivetotheantitumdrugs.ThisresearchsuggestsimptanceofODCintum