SDF-1αc-MYC蛋白调控速激肽受体-截短型mRNA表达的-挑战杯.DOCVIP

SDF-1α、c-MYC 蛋白调控速激肽受体-1mRNA表达的研究 The study of SDF-1α，c-MYC protein regulate tachykinin receptor 1 -trucated mRNA expression 【Abstract】 Objective : In order to investigate how SDF-1α and c-MYC protein regulates TACR1-trucated expression. Method: c-myc shRNA vector was constructed, small interfering RNA was employed for silencing c-myc gene in MCF-7 breast cancer cell. SDF-1α neutralized antibody was used in c-myc+cell group and c-myc- cell group, while other c-myc+ cell group and c-myc- cells group were cultured under normal condition. the mRNA level of TACR1-Trucated was determined by Real-Time PCR. Result: c-myc shRNA vector was constructed successfully, in the normal presence of SDF-1α, the level of TACR1-Tr mRNA in c-myc- cell group were lower than that in c-myc+cell group（P0.05）. But in the presence of SDF-1α neutralized antibody ,TACR1-Tr mRNA level of c-myc- cell group was higher than that of c-myc+ cell group（P0.05）. Conclusion: in the normal culture condition, c-MYC protein may transactivate TACR1-Tr transcription in MCF-7 cell; in the presence of SDF-1α neutral antibody, c-MYC protein lost the activity of transactivating for TACR1-Tr transciption. 【Key words】stromal cell derived factor 1α; tachykinin receptor1; shRNA; c-myc 【前言】前速激肽原PPT-I 是高度保守的单拷贝基因，通过替换剪切和转录后修饰产生几个属于速激肽(tachykinins)家族的肽，如P物质和神经激肽A等，并以不同的亲和性结合三种受体（tachykinin receptor -1,2,3）[1]。PPT-I 肽与神经内分泌系统有关联，在中枢神经系统和一些肿瘤组织呈构成性表达，也有研究发现骨髓基质细胞表达PPT-I肽，调节骨髓造血活动[2-4]，其中P物质和神经激肽A等作为中间递质参与机体多个器官功能活动[5,6]，在PPT-I 肽各类受体中，速激肽受体1 （tachykinin receptor 1, TACR1）在乳腺癌细胞骨转移过程中扮演极为重要的角色，TACR1分为全长型(TACR1-Fl)和截短型(TACR1-Tr)两种受体，属于G蛋白偶联7次跨膜受体。在乳腺癌细胞中，PPT-I基因过表达常导致TACR1-Tr以及几个相应的细胞因子产生，而在正常乳腺上皮细胞TACR1-Tr 表达量低，甚至难以检测，而TACR1-Fl表达量比较高，在恶性度比较高的乳腺癌细胞, 情况刚好相反，TACR1-Tr 受体表达上调是乳腺癌细胞获得侵袭性的特征之一，尤其是获得骨转移特性[7-9]。 基质细胞衍生因子1α（stromal derived cell factor 1α, SDF-1α）又称CXCL12或前B细胞刺激因子，与趋化因子受体4（CXCR4）形成CXCL12-CXCR4生物学轴，通过细胞内信号传导作用，调节相关转录蛋白，同样也调节TACR1受体表达[10],我们前期实验也表明SDF-1α表达水平与TACR1-Tr转录水平成负相关。 许多肿瘤的发生都与c-myc基因的过度表达和扩增有关，在一部分乳腺癌病例中c-myc