rhddADAM15抑制肝癌细胞Bel-7402增殖的作用研究-论文.pdf

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1、中国细胞生物学学报ChineseJoumalofCellBiology2014，36(4)：4l5_420DOI：10．11844／~cb．2014．04．0421rhd15抑制肝癌细胞Bel-7402i~殖的作用研究侯颖储敏王晓敏陈蕴金坚(江南大学药学院，药物设计与分子药理实验室，无锡214122)摘要ADAM15属于跨膜蛋白ADAM家族中的一员，在乳腺癌、宫颈癌、卵巢癌等多种实体瘤中均发现其表达量提高。ADAM15在降解细胞外基质、介导细胞的黏附、细胞间信号转导及在肿瘤发展进程中起到重要作用。因其去整合素区域含有RGD序列，ADAM15可与多种整合素相互作用。在前期工作中，实

2、验组利用大肠杆菌表达系统表达了重组．&ADAM15去整合素区域蛋白，记作rhddADAM15。该研究将进一步针对rhddADAM15抑制肝癌细胞Bel一7402增殖的机理进行分析及探讨。SRB法显示，rhddADAM15可抑制Bel一7402细胞增殖并呈剂量依赖性，IC50为1．14gmol／L；利用DAPI核染发现，rhddADAMl5可显著诱导Be1．7402~胞凋亡；流式细胞仪分析发现，rhddAD—AM15浓度为61amol／L时，(87．44~7．25)％的细胞发生凋亡；PI单染分析细胞周期表明，rhddADAM15作用后，部分Bel一7402{~胞周期被阻滞于S期及G

3、2／M期，并呈剂量依赖性，4gmol／LrhddADAM15处理后G0／Gl期含量下降约14％；Westernblot分析显示，rhddADAM15可下调Bel一7402细胞周期蛋白CDC2a~表达，抑制CDC2．Tyr的去磷酸化，引起G2／M期的阻滞。关键词ADAM15；去整合素；凋亡；CDC2EffectandMechanismofrhddADAM15ontheProliferationofBe1．7402CellsHouYing，ChuMin，WangXiaomin，ChenYun，JinJianabofDrugDesignandMolecularPharmacology,

4、SchoolofPharmaceuticalSciences，JiangnanUniversity,Wuxi214122,China)AbstractADAM15，amemberoftransmembraneproteinsADAMs，isover-expressedinmanysolidtumorssuchasbreast，colorectalandovariancancer,playingimportantrolesinthedegradationofextracellularmatrix，celladhe—sion，intracellularsignaltransducti

5、onandpathologicalchangesintumors．BecauseoftheRGDmotifinitsdisintegrindomain．ADAM15isconsideredtointeractwimintegrinsmultifariously．WehaveestablishedtherecombinanthumandisintegrindomainofADAM15(rhddADAM15、byE．coliinourpreviousresearch．111isstudyaimedatassessingtheef-feetandmechanismofrhddADAM1

6、5ontheproliferationofBel一7402cells．rhddADAM15inhibitedtheproliferationofBel一7402withanIC50of1．14gmoFLbySRBassay．Bel一7402cellshadapoptoticamorphologicalnucleuschangesusingDAPIstaining，and(87．44~7．25)％ofBel-7402cellsshowedapoptoticfeatureswhentreatedwith6~maol／LrhddAD—AM15analyzedbyPIstaining．M

7、oreove~partialSandG2／MarrestswereobservedonBel一7402cells．ⅥentreatedwimrhddADAM15attheconcentrationof4rmaol／L，theGdG1phaseofBe1．7402cellsreducedby14％．ThelevelofCDC2wasdown—regulatedandthephosphorylationofCDC—TyrWasincreasedwhicharousedtheG2／Ma