1、華中科技大學(xué)碩士學(xué)位論文雷帕霉素抑制人骨肉瘤MG63細(xì)胞周期及CyclinD1表達(dá)的實(shí)驗(yàn)研究姓名：高翔申請(qǐng)學(xué)位級(jí)別：碩士專(zhuān)業(yè)：外科學(xué)（骨科）指導(dǎo)教師：楊述華201004AbstractObjective：ToinvestigatetheinhibitiveeffectsofcellcycleCyclinD1expressionbyrapamycininMG63cellsprobethepossiblemechanism.Method：M

2、G63cellswereculturedinvitrotreatedwithdifferentconcentrationofrapamycin(1nmolL、10nmolL、25nmolL).UsedCCK8bywatersolubletetrazoliumsalt(WST8)assaytodetectthegrowthinhibitiveeffectsinMG63cellstreatedwithrapamycin；Flowcytome

3、trywasusedtoanalysisMG63cellcycle；ThegeneexpressionofCyclinD1wasexaminedbyRTPCRimmunocytochemistrySABC.Result：CCK8assayshowedthatthegrowthofMG63cellswassignificantlyinhibitedbyrapamycinitrevealedatimedependentrelationshi

4、p(P＜0.05).Thelongertherapamycintreatedthehigherthecellproliferationinhibitionrateperfmed.Buttherewasnosignificantlydifferentbetweenthreerapamycingroups(P＞0.05)；FlowcytometricanalysisrevealedasignificantlyG0G1phasearresti

5、nMG63cellstreatedwithrapamycin(P＜0.05).TheG0G1phasecellsweresignificantlyincreasedinthreerapamycingroupscomparedwiththecontrolgrouptheSphasecellsweresignificantlydecreased；RTPCRtestshowedtherewasnosignificantchangebetwee

6、ndifferentrapamycingroupscontrolgroupintheexpressionofCyclinD1mRNA(P＞0.05).CyclinD1mRNAexpressionwasnotsignificantlyaffectedbyrapamycininMG63cellsImmunocytochemistryanalysisshowedasignificantdownregulationofCyclinD1prote

7、ininrapamycingroupsitrevealedaconcentrationdependentrelationship(P＜0.05).Conclusion：RapamycincaninhibitthegrowthcellcycleofMG63cellsdownregulatetheexpressionofCyclinD1proteinbutithadnosignificanteffectintheexpressionofCy