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1、第42卷第5期浙江大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)Vol42No52013生JOURNALOFZHEJIANGUNIVERSITY(MEDICALSCIENCES)2013http:www.journals.zju.edu.cn/medDOI:10.3785/j.issn.1008-9292.2013.05.005木犀草素對(duì)人肝癌細(xì)胞HepG2的促凋亡作用王遠(yuǎn)鵬,周亮,龔興國(guó)(浙江大學(xué)生命科學(xué)學(xué)院生物化學(xué)所,浙江杭J,I、I310058)[摘要]目的:研究木犀草素(Luteolin)對(duì)人肝癌細(xì)胞HepG2的生長(zhǎng)抑制與促凋亡作用機(jī)制。方法:用梯度濃度木犀草素作用于多種腫瘤細(xì)胞,以MT
2、T法檢測(cè)藥物對(duì)細(xì)胞的生長(zhǎng)抑制作用,以流式細(xì)胞術(shù)檢測(cè)HepG2細(xì)胞內(nèi)活性氧水平,以AnnexinVFITC/PI雙染法利用流式細(xì)胞術(shù)檢測(cè)HepG2細(xì)胞凋亡情況,以Western.blot檢測(cè)HepG2細(xì)胞凋亡相關(guān)通路蛋白表達(dá)水平。結(jié)果:木犀草素對(duì)多種腫瘤細(xì)胞的生長(zhǎng)具有濃度和時(shí)間依賴的抑制作用(72h最高抑制率超過(guò)60%),并能有效降低HepG2細(xì)胞內(nèi)活性氧水平(約降低41.11%)。木犀草素作用下HepG2細(xì)胞凋亡率可達(dá)14.43%左右。木犀草素能夠降低HepG2細(xì)胞內(nèi)iASPP蛋白表達(dá)水平并上調(diào)p53與ASPP2蛋白表達(dá)。結(jié)論:木犀草素能夠抑制腫瘤細(xì)胞生長(zhǎng),降低Hep
3、G2細(xì)胞內(nèi)活性氧水平,并調(diào)節(jié)細(xì)胞內(nèi)相關(guān)凋亡通路蛋白水平,從而誘導(dǎo)HepG2細(xì)胞凋亡。[關(guān)鍵詞]木犀草素;癌,肝細(xì)胞;肝腫瘤;細(xì)胞凋亡;活性氧;腫瘤抑制蛋白質(zhì)p53;流式細(xì)胞術(shù);腫瘤細(xì)胞,培養(yǎng)的[中圖分類號(hào)]R34[文獻(xiàn)標(biāo)志碼]A[文章編號(hào)]1008-9292(2013)05—0504—07Pro-apoptoticeffectsofluteolinonhepatomaHepG2cellsWANGYuanpeng,ZHOULiang,GONGXingguo(InstituteofBiochemistry,CollegeofLifeSciences,ZhejiangUni
4、versity,Hangzhou310058,China)lAbstract]Objective:ToinvestigatetheeffectofluteolinoncellgrowthandapoptosisofHepG2cellsnvitro.Methods:CulturedHepG2,HL60,A549andL02cellsweretreatedwithluteolinfordifferentdoses(0ml,2.5ml,10g/mland20g/m1)andvariedtimes(0h,24h,48hand72h).Cellviabilitywasmeasu
5、redwithMTI"assayandICwascalculated.Thereactiveoxygenspecies(ROS)levelsinHepG2cellstreatedwithluteolinfor6hand12hweremeasuredwithflowcytometry.CellapoptosisofHepG2cellstreatedwithluteolinfor24hwasexaminedwithflowcytometryandAnnexinV—FITC/PI.Expressionlevelsofapoptosispathwayproteins(p53,
6、ASPP2andiASPP)inHepG2cellsweredetectedwithwesternblotandthedoseandtime.effectwasanalyzed.Results:Luteolineffeetivelyinhibitedtumorcellproliferationinadose.a(chǎn)ndtime—dependentmanner,andtheinhibitionratesof20LLmlLuteolinfor72hwere39.34%,62.90%.57.57%and62.90%toL02,HepG2,HL60andA549cells,res
7、pectively.TheintracellularROSlevelwasdecreasedinHepG2cellsby13.88%and41.11%收稿日期:2012-09.14修回日期:2013-04-01基金項(xiàng)目:國(guó)家自然科學(xué)基金(30900760);浙江省科技計(jì)劃項(xiàng)目(2006C12084).作者簡(jiǎn)介:王遠(yuǎn)鵬(1986一),男,碩士研究生,主要從事信號(hào)傳導(dǎo)通路和作用機(jī)制研究;E-mail:wangyuanpeng1986@yahoo.con.cn通訊作者:龔興國(guó)(1955一),男,研究員,主要從事重要藥用蛋白和臨床診斷用酶的基因工程研究;E-