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(<中国癌症亲志》2011年第2l 卷第6期CH IN A O N CO LO G Y201lV01．21 N o．642l三苯氧胺诱导人乳腺耐药及自噬的癌M CF一7细胞株关系研究[ 摘要]背景与目的：三苯氧胺( tam oxi fen) 作为第一代选择性雌激素受体调节剂( sel ecti ve estrogenm odul ator，SERM ) 被广泛地应用于激素敏感型乳腺癌的内分泌一线治疗。三苯氧胺耐药的发生严重receptor限制了临床治疗，是乳腺癌患者用药面临的重大难题，明确其耐药机制对乳腺癌的治疗有重要临床意义。本研究通过体外诱导人乳腺癌细胞M CF-7三苯氧胺耐药，探讨细胞产生三苯氧胺耐药时自噬水平的变化与M APK家族蛋白细胞外信号调节激酶( extracel l ul arsi gnal ～regul ated ki nase，ERK) 蛋白表达量及磷酸化水平的变化。方法：浓度递增筛选法诱导M CF一7细胞耐药，透射电镜观察M CF一7细胞与耐药细胞内的自噬泡数量，CCK8法检测细胞增殖状态，应用W estern bl ot检钡t]LC3II、ERKl ／2、Phospho—ERKl ／2蛋白的表达情况。结果：诱导的三苯氧胺耐药细胞株TR5达到5 um ol ／L的耐药浓度。TR5细胞内的自噬泡数量与LC3II表达量明显高于M CF一7细胞。ERK蛋白在两种细胞中的表达量差异无统计学意义，但其在TR5中的磷酸化水平bEM CF-7细胞高。结论：M CF一7三苯氧胺耐药细胞有较高的自噬水平，M APK信号通路参与了三苯氧胺耐药。[关键词】乳腺癌：三苯氧胺；M CF一7细胞；自噬DO I：10．3969／j ．i ssn．1007—3969．2011．06．001中图分类号：R737．9文献标志码：A文章编号：1007-3639( 2011) 06-0421-06Studi es on resi stant i nducti on of breast cancer cel l M CF．7 to tam oxi f en i n vi troM AXi ao- yu,of BreastLIU zhe—bl n, YU San—j i an．LI Shuang, H O U Y1- f eng, s}弧0 Zhi - m i n( D epartm entSurgery, CancerC011ege, FudanU ni verslty, ShanghalH ospi tal , FU dan U ni versi ty：D epartm entof O ncol ogy, SbanghalM edl cal200032, Chi na)Correspondence[ Abstract]totH O UYi - f engE- m ai l - yi f enghoupi tt#yahoo．CO BBackgroundandpurpose：Tam oxi fen( TAM ) ，the pi oneeri ng sel ecti veestrogen receptorm odul ator( SERM ) ，w hi chbl ocksestrogenacti onby bi ndi ngto theER i n breastcancers，has been usedubi qui tousl yforendocri netherapyfor the horm one- sensi ti vebreast Cal ICCT．Intri nsi c andacqui redresi stance to TAMl i m i ts the cl i ni caluse of TAM to anal TO Wtherapeuti cw i ndowandtheyarebi g chal l engesto thedrug therapyforbreast cancer pati ents．Better understandi ngof the TfⅢresistant m echani sm si s therefore of consi derabl e cl i ni cal si gni fi cance．Inourstudy,w e i nduced breast cancer cel l M CF．7 resi stant to TAMi n vi tro and tri edtoexpl orethechanges of autophagi c Ievel s andtheexpressi onof ERK( extracel l ul ar si gnal -regul atedki nase，oneof M APKfam i l y protei ns) and Phospho- ERKl ／2i nTAMresi stant cel l scom paredw i th M CF- 7cel l s．M ethods：Stepw i se TAMsel ecti on W aS used to establ i sh the TAMresi stant TR5subl i ne．Autophagi cvacuol es i n cel l s w ereobservedby m eans of transm i ssi on el ectronm i croscopy．nl egrow thof the t W Ocel lw ere m eaSuredbyW esternblot．Results：The删resistant eel l l i ne TR5 w e establ i shed Can be resi stant to 5呦ol ／LT久M ．111enum ber ofautophagi c vacuol es and theexpressi onofLC3 II protei ni n TR5 w ereobvi ousl y hi gher thanthat i n M CF-7．There W aS nosi gni fi cantdi fference i n theprotei n expressi onl evel of ERKl ／2 betw een M CF一7 andTR5l i nes w ere m easuredby CCK8，andtheexpressi onof LC3II，ERKl ／2andPhospho—ERKl ，2cel l s，but the l evel ofPhospho—ERKI／2w asm arkedl y hi gheri n TR5 cel l s than thatTAMresi stant M CF一7 cei l s haverel ati vel y hi gh autophagi cl evel and～p—Kpathw ayi s i nvol ved i nfaci l i tati ngTAMi n M CF- 7 cei l s．Concl usi on：resi stance．基金项目：上海市科委浦江人才计划( N o：09PJ l 402700) ；国家自然科学基金( N o：81072165) 。通信作者：侯意枫E—m ai l ：yi fenghoupi tt@ yahoo．tom万方数据