浅蓝菌素对人胆管癌细胞株rbe生长的影响及机制的实验分析-消化病学专业论文 word格式.docxVIP

Thestudies to effectofceruleninonhuman cholangiocarcinomacelllineRBE anditsmechanismsAbstractObjective:Tostudytheeffectofceruleninonthegrowthandapoptosisofhuman cholangiocarcinomacancercelllineRBEinvitro，andtoexploreits possiblemechanisms.Methods:RBEcellswereculturedroutinelyinvitro,FollowingexposuretoCerat differentconcentrationsindifferenttimes,ThenRBEcellswereharvestedand investigated.Thegrowthof concentrationscancercellsRBEwasevaluatedby MTTassay, andinhibitionratioswerecalculatedinallgroupsandsuitableIC50ofceruleninwereworkedoutbycalculatation.Withusingdifferentconcentrationscerulenin(0、5、10、20ug/ml)todealwithRBE,theapoptosis ratioes andchanges ofcellcycleweredetectedby flowcytometryandAnnexinV/PIfluosstainging,andtheexpressionofFASmRNA、Bcl-2mRNA、BaxmRNAweredetectedbyRT-PCR,Bcl-2、BaxproteinumweredetectedbyWesternBlotinvitro.Results：(1)Cerulenincouldinhibitthegrowthofthehumancholangiocarcinoma carcinomacelllineRBE,andtheinhibitoryeffecthaddose-and-timedependence(P0.05). (2)TheearlystageapoptosisandcyclomorphosisofRBE cellswasdetectedbyflowcytometryandAnnexinV/PIfluosstaingingaftertheywereincubated48hourswith cerulenin .With the increase of Cer concentration, ratio of S phase increasedfrom(6.97±0.37)to(26.43±1.38)andG2phasedeclinedhomologously（P0.05）,TheearlystageapoptosisrateofRBEcellswas(10.73±0.82)%whentheywereincubatedwith 5ug/mlCerandtheratewas(45.1±1.73)%with20ug/mlCer.Appearantly,apoptosisrate waspositively correlatedtoCerconcentration.(3)TheexpressionofFASmRNAand Bcl-2mRNAdecreasedwithconcentration-dependenceafterRBEcellswereincubated48hourswithCer,whichwasdetectedbyRT-PCR.Otherwise,TheexpressionofBaxmRNA increased with concentration-dependence, Comparisons of all Cer groups and controlgrouphavesignificantdifference（P0.05）.(4)TheexpressionofBcl-2was downregulatedandtheexpressionofbaxwasupregulatedsignificantlyafterRBEcellswereincubated48 hourswithCer,whichwasdetectedbyWesternBlot.AndtheratioofBcl-2/Baxwere(4.29±0.15)、(1.82±0.02)、(1.01±0.08)、(0.