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1、GW3965在Raji细胞中对BLyS表达的影响黄艳1,黄刚2,胡长江2,曾益军2,许志臻2,陈岺曦2,何凤田2,宋方洲1(400016重庆,重庆医科大学分子医学与肿瘤研究中心1;400038重庆,第三军医大学基础医学部生物化学与分子生物学教研室2)[摘要]目的探讨肝X受体(liverXreceptors,LXRs)的特异性激动剂GW3965在Raji细胞中对B淋巴细胞刺激因子(Blymphocytestimulator,BLyS)表达的影响及可能机制。方法MTT法检测不同浓度的GW3965(浓度分别为0.5、5、10μmol/L
2、)对Raji细胞活性的影响,观察时间为加入GW3965共孵育后24h。用LXR的特异性激动剂GW3965刺激人B细胞淋巴瘤株Raji细胞,经RT-PCR检测LXR特异性靶基因三磷酸腺苷结合盒A1(ATP-bindingcassette,ABCA1)mRNA的表达;经RT-PCR和Westernblot检测BLyS的表达。结果对照组1‰DMSOD(492nm)为(0.632±0.055),0.5、5μmol/L和10μmol/L的GW3965D(492nm)为(0.609±0.073)、(0.612±0.052)和(0.628±0.
3、055)。所以LXR的特异性配体GW3965组对细胞活性无影响(P>0.05)。GW3965作用于Raji细胞后,ABCA1mRNA表达剂量依赖性的明显上调,表明LXR在Raji细胞中具有功能活性。LXR活化后可在转录和翻译水平剂量依赖性的明显抑制BLyS的表达。结论LXR在Raji细胞中可抑制BLyS的表达。[关键词]GW3965;B淋巴细胞刺激因子;Raji细胞[中图法分类号][文献标志码]A[基金项目]国家自然科学基金(81001302) [通信作者]宋方洲,E-mail:fzsongcq@163.comEffectofGW
4、3965onBLySexpressioninRajicellHuangYan1,HuangGang2,HuChangjiang2,ZengYijun2,XuZhizhen2,ChenLingxi2,HeFengtian2,SongFangzhou1(1MolecularMedicineandTumorResearchCenter,ChongqingMedicalUniversity,Chongqing400016,China;2DepartmentofBiochemistry&MolecularBiology,ThirdMilit
5、aryMedicalUniversity,Chongqing400038,China)[Abstract]ObjectiveToinvestigatetheeffectofliverXreceptor(LXR)specificagonistGW3965ontheexpressionofBlymphocytestimulator(BLyS)anditspotentialmechanism.MethodsEffectofvariousGW3965concentrationsonviabilityofRajicellswasdetect
6、edbyMTTtest.HumanBlymphocytecellsweretreatedwithLXRspecificagonistGW3965andthenABCA1(thespecifictargetgeneofLXR)mRNAwasdetectedbyRT-PCR;BLySmRNAandproteinwereassayedbyusingRT-PCRandWesternblot.ThepotentialbindingsiteofLXRintheBLySpromoterregionwasonlineanalyzed.Finall
7、y,determiningthelevelofLXRbyRT-PCR.Results Thelevelof1‰DMSOOD(492nm)levelwas(0.632±0.055),Thelevelof0.5μmol/L、5μmol/Land10μmol/LGW3965OD(492nm)were(0.609±0.073)、(0.612±0.052)和(0.628±0.055)。GW3965hadnoeffectontheviabilityofRajicells(P>0.05).ThelevelofABCA1mRNAwasincre
8、ased,treatedwithLXRspecificagonistGW3965withdifferentconcentrationsfor24hrespectivelyinRaji,whichdemonstratedLXRwasfunctiona