渥曼青霉素联合丝裂霉素对人肝癌hepg-2细胞增殖和凋亡的影响

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1、渥曼青霉素联合丝裂霉素对人肝癌HepG-2细胞增殖和凋亡的影响张健1，王洪林1，张潜1，赵媛2(400016重庆，重庆医科大学附属第一医院：肝胆外科1，眼科2)[摘要]目的研究渥曼青霉素联合丝裂霉素对肝癌HepG-2细胞增殖、凋亡及相关基因CyclinD1、Bad表达的影响。方法MTT比色法检测细胞增殖状态；Hoechst33258荧光染色观察凋亡细胞核形态学变化，流式细胞术检测肿瘤细胞周期和凋亡；逆转录聚合酶联反应及Westernblot检测PI3K/Akt信号传导通路相关基因和蛋白的表达。结果联合用药组的细胞增殖抑制率显著高于单独用药组(P<0.05)；联合

2、用药作用24h后流式细胞术显示细胞明显阻滞于G0/G1期；细胞凋亡率由处理前的（3.23±1.32）%提高到14.82±1.36）%(P<0.01)；荧光显微镜下可见典型的细胞凋亡形态学改变；联合用药后细胞p-Akt活化程度降低，CyclinD1表达量减少，Bad表达量增加（P<0.05)。结论渥曼青霉素与丝裂霉素联合应用，具有协同抗肿瘤效应。[关键词]肝癌；渥曼青霉素；丝裂霉素；凋亡；PI3K/Akt信号通路[中图法分类号]R735.7[文献标志码]AEffectsofWortmannincombinedwithmitomycinConHepG-2cellZh

3、angJian,1WangHong-lin,1ZhangQian1ZhaoYuan2（DepartmentofHepatobiliarySurgery,FirstAffiliatedHospital,ChongqingMedicalUniversity,Chongqing400016.2DepartmentofOphthalmology,FirstAffiliatedHospital,ChongqingMedicalUniversity,Chongqing400016.）Abstract:Objectivethisstudywastoinvestigatethe

4、effectsofWortmannincombineduseofmitomycinContheproliferation,apoptosisandexpressionsofrelatedCyclinD1.Badgenesinlivercancer.MethodsCellsuppressionrateweredetectedbyMTTassay;ThenucleusapoptosisofHep-G-2cellswereexaminedbyHochest33258;ThechangeofcellcycleandapoptosisinHepG-2cellswasdet

5、ectedbyflowcytometry(FCM),expressionofPI3K/Aktsignalingtransductionpathwayrelatedgenesandproteinsweretestedbyreversetranscription-polymerasechainreaction(RT-PCR)andWesternBlotassay.ResultsTheinhibitionratioofcellproliferationwassignificanthigherinCombineduseofdrugsgroupsthanineithers

6、inglegroups(P<0.05);FCMshow:HepG-2cellswereprohibitedatG0/G1phrase,apoptosisratesofHepG-2cellsincreasedfrom（3.23±1.32）%to（14.82±1.36）%AfterincubationwithCombineduseofWortmannin、MMCfor24hours.(P<0.01).typicalchangesinapoptoticmorphologyofHepG-2cellswereobservedbyfluorescencemic-roscop

7、y.activationofp-Aktwasinhibited,mRNAandproteinlevelsofCyclinD1decreased,butBadincreased.ConclusionCombineduseofWortmannin、MMCshowedasynergicantineoplasticeffect.Keywords:livercancer,Wortmannin,mitomycinC(MMC),apoptosis,PI3K/Aktsignalingpathway通信作者：王洪林，电话：（023）89107534，Email；whlcqykd@

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