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1、302細胞與分子免疫學(xué)雜志(ChinJCellMollmmuno1)2015,3l(3·論著·文章編號:1007—8738(2015)03—0302—05漢黃芩素對膠質(zhì)瘤U87細胞增殖和侵襲的抑制作用及機制曾志青,劉洪(南華大學(xué)附屬第一醫(yī)院神經(jīng)外科,湖南衡陽421001)[摘要】目的探討漢黃芩素對膠質(zhì)瘤U87細胞增殖、侵襲的影響及其相關(guān)機制。方法U87細胞隨機分為對照組(加入2O的培養(yǎng)液)、(0、50、100)~moVL漢黃芩素處理組;細胞培養(yǎng)48h,應(yīng)用MIT法檢測細胞增殖情況,通過Transwell侵襲實驗檢測細胞侵襲能力,熒光實時定量PCR測定ezrinmR
2、NA表達,Westernblot法檢測ezrin、Bc1-2、Bax蛋白表達及ezrin蛋白磷酸化水平,用末端脫氧核糖核酸轉(zhuǎn)移酶介導(dǎo)的脫氧尿苷三磷酸缺口末端標(biāo)記法(TUNEL)測定腫瘤細胞凋亡指數(shù)(AI)。結(jié)果與0pjnol/L漢黃芩素組以及對照組相比,(50、100)m0L/L漢黃芩素組細胞增殖抑制率增加,且100~mol/L漢黃芩素組細胞增殖抑制率明顯高于50~moVL漢黃芩素組。隨著漢黃芩素濃度的增加,平均穿膜細胞數(shù)以及ezrinmRNA、ezrin蛋白、ezrin蛋白磷酸化水平、Bcl-2蛋白表達水平逐漸降低,而Bax蛋白表達水平、AI逐漸增加;0p~mo
3、l/L漢黃芩素組與對照組相比,無顯著性差異。結(jié)論漢黃芩素能夠減少膠質(zhì)瘤U87細胞增殖、侵襲,下調(diào)ezrin蛋白表達和磷酸化活性,并誘導(dǎo)細胞凋亡。[關(guān)鍵詞]漢黃芩素;膠質(zhì)瘤;ezrin蛋白;凋亡[中圖分類號]R285.5,R739.41,R392·33,Q279[文獻標(biāo)志碼]AInhibitoryeffectofwogoninontheproliferationandinvasionofglioblastomaU87cellsandrelatedmechanismZENGZhiqing,LIUHongDepartmentofNeurosurgery,F(xiàn)irstAff
4、iliatedHospital,UniversityofSouthChina,Hengyang421001,China[Abstract]ObjectiveToexploretheefectofwogoninontheproliferationandinvasionofglioblastomaU87cellsanditsrelatedmechanism.MethodsGlioblastomaU87cellswerecultured/nvitroinRPM11640mediumaddedwith1oomL/LfetaIbovineserum.Afterceladhe
5、rence,thecellswereinoculatedwith20gLculturesolution(controlgroup),20pLwogoninsolutions(0,50andlO0)pmol/Lfor48hours,respectively.The0pmoVLwogoningroupused20pLPBSinstead.CellproliferationwasanalyzedbyM1_rassay.ThecellinvasionabilitywasdetectedusingTranswelinvasionassay.Theexpressionsofe
6、zrinmRNAwasexaminedthroughrea1.timequantitativePCR.Theexpressionsofezrin。Bcl-2andBaxproteinsaswellasphosphorylatedezrinproteinlevelweremeasuredbyWesternblotting.Apoptosisindex(AI)wasdeterminedthroughterminaldeo×ynucleotidyltransferasemediateddeoxyuridinetriphosphatenickendlabeling(TUN
7、EL)assay.ResultsTheinhibitoryrateofcelproliferationsignificantlyincreasedin50and100vmoVLwogoningroupsascomparedwith0pmol/Lwogoningroupandcontrolgroup.Moreover,thatwashigherin100iJmoVLwogoningroupthan50pmoVLwogoningroup.Incomparisonwithcontroland0i~mol/Lwogoningroups,themeancellnumbers
8、ofper