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1、342細胞與分子免疫學雜志(ChinJCellMolImmune1)2015,31(3·論著·文章編號:1007—8738(2015)03—0342—04黃芩素對骨肉瘤細胞增殖和侵襲的抑制作用及機制唐志斌,李春,陳志偉(南華大學第一附屬醫(yī)院骨外科,湖南衡陽421001)【摘要]目的探討黃芩素對骨肉瘤細胞增殖、侵襲的影響及其相關機制。方法以骨肉瘤MG-63細胞為研究對象,分別加入20的培養(yǎng)液(對照組)或(0、100、200)~moVL黃芩素溶液處理細胞48h,應用MTF法分析細胞增殖情況,通過Transwell侵襲實驗檢測細胞侵襲能力,熒光
2、實時定量PCR測定ezrinmRNA表達,采用Westernblot法檢測ezrin蛋白及磷酸化ezrin(p-ezrin)蛋白表達,利用原位末端標記法(TUNEL)測定腫瘤細胞凋亡指數(shù)(AI)。結(jié)果(100、200)~moVL黃芩素組細胞增殖抑制率較0moL/L黃芩素組升高,且200~moVL黃芩素組細胞增殖抑制率明顯高于100~moVL黃芩素組。隨著黃芩素濃度的增加,平均穿膜細胞數(shù)以及ezrinmRNA、ezrin蛋白、p-ezrin蛋白表達水平逐漸降低,而AI逐漸增加,與對照組和0~snol/L黃芩素組比較,差異均有顯著降低,而0p
3、xaoVL黃芩素組與對照組相比無明顯變化。結(jié)論黃芩素能夠抑制骨肉瘤MG-63細胞增殖、侵襲,其機制可能與抑制ezrin蛋白表達和活性及促進腫瘤細胞凋亡有關。[關鍵詞]黃芩素;骨肉瘤;ezrin蛋白;凋亡[中圖分類號]R392.12,Q279,R285.5[文獻標志碼]AInhibitoryefectofbaicaleinontheproliferationandinvasionofosteosarcomacellsandmechanismTANGZhibin,LIChun,CHENZhiweiDepartmentofOrthopedics
4、,F(xiàn)irstAfiliatedHospital,UniversityofSouthChina,Hengyang421001,ChinaIAbstractIObjectiveToexploretheefectofbaicaleinontheproliferationandinvasionofosteosarcomacellsandjtsrelatedmechanism.MethodsOsteosarcernaMG-63cellsthatwerecultured/nv/trowererespectivelytreatedwith20uLcul
5、turemedium(controlgroup),dehydratedalcohol(0wmol/Lbaicaleingroup),100and200pmoVLbaicaleinsolutionfor48hours.CelIproliferationwasanalyzedbvM1_rassay.ThecelIinvasionabilitywasdetectedusingTranswelI?invasionassay.TheexpressionofezrinmRNAwasexaminedbyreal—timequantitativePCR.
6、Theexpressionsofezrinproteinandp-ezrinproteinweremeasuredusingWesternblotting.Apeptosisindex(AI)wasdeterminedbyterminaldeoxynucle~idyltransferase-mediateddUTPnickendIabeling(TUNEL).ResultsTheinhibitoryratesofcelIproliferationsignificantlyincreasedin100and200iJmol/Lbaicale
7、ingroupsascomparedwith0pmol/Lbaicaleingroup.Moreover,thatwashigherin200pmol/Lbaicaleingroupthanin100wmol/Lbaicaleingroup.Incomparisonwithcontroland0wmol/Lbaicaleingroups,themeancellnumbersofpermeatedmembraneandlevelsofezrinmRNA.ezrinproteinandP-ezrinproteingraduallydecrea
8、sed.butA1wasgraduallyelevatedwiththeincreaseofbaicaleinconcentrations.whereastherewasnosignifica