Y盒结合蛋白1对白血病K562细胞多药耐药基因1表达调控.docVIP

Y盒结合蛋白1对白血病K562细胞多药耐药基因1表达调控 　【摘要】 观察多柔比星诱导K562细胞多药耐药基因1（mdr1）基因表达过程中Y盒结合蛋白1（YB1）表达和核易位的变化情况，初步探讨YB1蛋白对mdr1基因的转录调控。方法： 多柔比星间歇性长期作用于K562细胞，剂量逐渐增加。RTPCR检测mdr1，YB1基因表达，流式细胞仪检测mdr1基因编码的P糖蛋白（Pgp）表达，蛋白质印迹检测YB1蛋白核易位。通过RNA干扰技术使K562细胞中YB1基因表达沉默，多柔比星处理YB1基因沉默细胞，RTPCR、流式细胞仪分别检测mdr1 mRNA和Pgp的表达。结果： 经多柔比星作用后K562细胞mdr1基因转录上调，Pgp表达增加，同时YB1基因转录也增强，且出现明显的核易位。YB1基因沉默后，mdr1基因诱导性表达减少。结论： 多柔比星诱导K562细胞mdr1基因表达的过程中，YB1对于mdr1基因的转录活化起一定的作用。 【关键词】 Y盒结合蛋白1 多药耐药基因1 多药耐药 转录调节 ［Abstract］ Objective: To investigate the effect of YB1 on the transcription of mdr1 gene.Methods： K562 cells were treated with doxorubicin (DOX)at different concentrations and times.Expression of mdr1 and YB1 genes were examined by reverse transcriptionpolymerase chain reaction (RTPCR).Pglycoprotein (Pgp)was detected by flow cytometry.Cyto/nuclear protein was extracted for Western blotting to detect YB1.To inhibit the expression of YB1 gene in K562 cell using RNA interference (RNAi)targeting YB1 gene, then detect the expression of mdr1 and Pgp in YB1 gene silenced cells after treated with DOX. Results： When K562 cells were exposured to DOX, the mdr1 gene was highly expressed as well as its corresponding protein Pgp.Further, DOX upregulated the expression of YB1 gene, and promoted YB1 protein translocate to nucleus.Another, when YB1 gene was silenced, the expression of mdr1 gene and Pgp were obviously decreased in K562 cells treated with DOX. Conclusion： Doxorubicin could induce the expression of mdr1 gene in K562 cells, it may be due to YB1 activating the transcription of mdr1 gene. ［Key words］ YB1; mdr1; multidrug resistance; transcriptional regulation 多药耐药（MDR）是白血病化疗失败和复发的主要原因，多药耐药基因1(mdr1)及其编码的蛋白P糖蛋白（Pglycoprotein, Pgp）是导致耐药产生的最主要原因。mdr1基因转录调控机制比较复杂, 多种转录因子和阻遏蛋白参与了mdr1基因的表达调控。Y盒结合蛋白1 (Ybox binding protein1, YB1)是Y盒蛋白家族成员之一，其结构中高度保守的冷休克区（cold shock domain, CSD）可识别多种基因启动子区的反向CCAAT盒，即Y盒，并与之特异性地结合。mdr1基因启动子区域含有CCAAT序列，有关白血病中YB1调控mdr1基因表达的研究报道尚较少见，我们通过多柔比星诱导K