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1、Survivin反義寡核苷酸對(duì)骨肉瘤細(xì)胞凋亡的影響腫瘤2006年6月第26卷第6期TumorVo1.26,No.6,Jun.2006?533?Survivin反義寡核苷酸對(duì)骨肉瘤細(xì)胞凋亡的影響梁新軍,夏仁云(華中科技大學(xué)同濟(jì)醫(yī)學(xué)院附屬同濟(jì)醫(yī)院骨科,武漢430030)BasicResearch?基礎(chǔ)研究[摘要]目的:采用survivin反義寡核苷酸(ASODN)封閉骨肉瘤細(xì)胞survivin的作用,觀察其對(duì)骨肉瘤細(xì)胞系MG-63增殖和凋亡的影響.方法:以脂質(zhì)體為載體,介導(dǎo)survivin反義寡核苷酸轉(zhuǎn)染人骨肉瘤細(xì)胞系MG-6
2、3,用MTT法測定細(xì)胞增殖;流式細(xì)胞儀檢測轉(zhuǎn)染survivin反義寡核苷酸后細(xì)胞凋亡率;Westernblot及半定量RT—PCR檢測轉(zhuǎn)染survivin反義寡核苷酸后的骨肉瘤細(xì)胞survivin蛋白及mRNA表達(dá).結(jié)果:脂質(zhì)體介導(dǎo)survivin反義寡核苷酸轉(zhuǎn)染骨肉瘤細(xì)胞系MG-63后,細(xì)胞增殖明顯受到抑制,細(xì)胞凋亡率明顯增加,骨肉瘤細(xì)胞survivin蛋白及mRNA表達(dá)均明顯降低.結(jié)論:脂質(zhì)體介導(dǎo)轉(zhuǎn)染sur—vivin反義寡核苷酸可以抑制細(xì)胞增殖,有效降低細(xì)胞內(nèi)survivin基因的表達(dá),促進(jìn)細(xì)胞凋亡.[關(guān)鍵詞]骨肉瘤
3、;生存素;寡核苷酸類,反義;細(xì)胞凋亡[中圖分類號(hào)]R738.1;R7336[文獻(xiàn)標(biāo)識(shí)碼]A[文章編號(hào)]10007431(2006)06—0533—04Antisense0lig0nucle0tidetargetingsurvivingeneinducesceilapoptosisofosteosarcomaLIANGXin—Jun,XIARen—Yun(DepartmentofOrthopaedics,TongjiHospital,TongjiMedicalCollege,HuazhongUniversityofScien
4、ceandTechnology,Wuhan430030,China)[ABSTRACT-]0bj}ective;Theaimofthisstudywastoinvestigatetheeffectofantisenseoligonucleotide(ASODN)targetingsurvivinonproliferationandapoptosisofosteosarcomacellslineMG-63.Methods:SurvivinASODNwasdesignedandtransfectedintoMG--63cell
5、smediatedbyliposomecarrier.Culturedcellsweredividedinto5groups:controlgroup,lipofectiongroup,senseoligonucleotide(SODN)group,mismatcho¨gonucleotide(MODN)group,andASODNgroup.Proliferationofovariancellswasdetectedbymethylthiazolyltetrazolium(MTT)assay;apoptoticratew
6、asdeterminedbyflowcytometry;survivinproteinandmRNAexpressionweremeasuredbywesternblotandreversedtranscriptasepolymerasechainreaction,respectively.Results:TheproliferationofMG-63cellswasinhibitednotablyafterASODNtransfectionandtheapoptosisrateofASODNgroupswassignif
7、icantlyhigherthanthatofothergroups.SurvivinproteinandmRNAexpressioninASODNgroupweresignificantlyde—creasedcomparedwiththatinthecontrolgroups.Conclusions:Liposome-mediatedsurvivinASODNtransfectionsignificantlyin-hibitstheproliferationofosteosarcomaMG-63cells,reduce
8、stheexpressionofsurvivingenes,andinducescellapoptosis.[KEYWORDS]Osteosarcoma;Survivin;oligodeoxynucleotide,antisense;ApoptosisrTumor,2006,26(6):533—536]
