YB1 regulates miR-205/200b-ZEB1 axis by inhibiting microRNA maturation in hepatocellular carcinoma.
YB1 regulates miR-205/200b-ZEB1 axis by inhibiting microRNA maturation in hepatocellular carcinoma.
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DOI：
10.1002/cac2.12164
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发表时间：
2021-07
期刊：
影响因子：
--
通讯作者：
Piao HL
中科院分区：
文献类型：
--
作者：
Liu X;Chen D;Chen H;Wang W;Liu Y;Wang Y;Duan C;Ning Z;Guo X;Otkur W;Liu J;Qi H;Liu X;Lin A;Xia T;Liu HX;Piao HL
Y-box结合蛋白1(YB1或YBX1)在肿瘤的发生、发展中起重要作用。然而，YB1是否通过调节非编码RNA影响恶性转化仍在很大程度上尚不清楚。本研究旨在研究YB1与microRNAs的关系，揭示YB1通过miRNAs介导的调控网络影响肿瘤恶性的潜在机制。通过细胞增殖、创面愈合和跨孔侵袭实验研究YB1在肝癌细胞中的生物学功能。应用基因芯片技术筛选出YB1基因在肝癌细胞系中表达异常的miRNA。采用实时定量聚合酶链式反应、双荧光素酶报告基因检测、RNA免疫沉淀和下拉实验等方法检测YB1对miR-205和miR-200b的调控作用。通过下拉实验和免疫共沉淀实验确定了YB1、DgCr8、DICER、TUT4和TUT1之间的关系。免疫荧光染色检测YB1、DgCr8和DICER的细胞共定位。通过尾静脉注射转导YB1 shRNA或shControl的MHCC97H细胞，观察YB1对裸鼠肿瘤转移的影响。免疫印迹和免疫组织化学方法检测上皮向间充质转化标志物的表达水平。YB1通过对miR-205/200b-ZEB1轴的部分调控，不依赖蜗牛，促进肝癌细胞迁移和肿瘤转移。YB1通过保守的冷激波结构域与微处理器DgCr8和Dird以及TUT4和TUT1相互作用，抑制miR-205和miR-200b的成熟。随后，miR-205和miR-200b的下调增强了ZEB1的表达，从而导致细胞迁移和侵袭的增加。此外，对肝细胞癌和正常肝组织基因表达数据的统计分析表明，YB1的表达与ZEB1的表达呈正相关，且与临床预后显著相关。这项研究揭示了YB1通过调节肝癌细胞中的miR-205/200b-ZEB1轴促进癌症进展的先前未被描述的机制。此外，这些结果表明YB1可能通过miRNAs介导的基因调控发挥生物学功能，可以作为人类癌症的潜在治疗靶点。YB1通过抑制microRNA成熟来调节miR-205/200b-ZEB1轴。
Y‐box binding protein 1 (YB1 or YBX1) plays a critical role in tumorigenesis and cancer progression. However, whether YB1 affects malignant transformation by modulating non‐coding RNAs remains largely unknown. This study aimed to investigate the relationship between YB1 and microRNAs and reveal the underlying mechanism by which YB1 impacts on tumor malignancy via miRNAs‐mediated regulatory network. The biological functions of YB1 in hepatocellular carcinoma (HCC) cells were investigated by cell proliferation, wound healing, and transwell invasion assays. The miRNAs dysregulated by YB1 were screened by microarray analysis in HCC cell lines. The regulation of YB1 on miR‐205 and miR‐200b was determined by quantitative real‐time PCR, dual‐luciferase reporter assay, RNA immunoprecipitation, and pull‐down assay. The relationships of YB1, DGCR8, Dicer, TUT4, and TUT1 were identified by pull‐down and coimmunoprecipitation experiments. The cellular co‐localization of YB1, DGCR8, and Dicer were detected by immunofluorescent staining. The in vivo effect of YB1 on tumor metastasis was determined by injecting MHCC97H cells transduced with YB1 shRNA or shControl via the tail vein in nude BALB/c mice. The expression levels of epithelial to mesenchymal transition markers were detected by immunoblotting and immunohistochemistry assays. YB1 promoted HCC cell migration and tumor metastasis by regulating miR‐205/200b‒ZEB1 axis partially in a Snail‐independent manner. YB1 suppressed miR‐205 and miR‐200b maturation by interacting with the microprocessors DGCR8 and Dicer as well as TUT4 and TUT1 via the conserved cold shock domain. Subsequently, the downregulation of miR‐205 and miR‐200b enhanced ZEB1 expression, thus leading to increased cell migration and invasion. Furthermore, statistical analyses on gene expression data from HCC and normal liver tissues showed that YB1 expression was positively associated with ZEB1 expression and remarkably correlated with clinical prognosis. This study reveals a previously undescribed mechanism by which YB1 promotes cancer progression by regulating the miR‐205/200b‒ZEB1 axis in HCC cells. Furthermore, these results highlight that YB1 may play biological functions via miRNAs‐mediated gene regulation, and it can serve as a potential therapeutic target in human cancers. YB1 regulates miR‐205/200b‐ZEB1 axis by inhibiting microRNA maturation.
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DOI：
10.1083/jcb.201110008
发表时间：
2012-04-16
期刊：
The Journal of cell biology
影响因子：
--
作者：
Kawai S;Amano A
通讯作者：
Amano A
影响因子：
64.5
作者：
Cancer Genome Atlas Research Network. Electronic address: wheeler@bcm.edu;Cancer Genome Atlas Research Network
通讯作者：
Cancer Genome Atlas Research Network
影响因子：
14.9
作者：
Colaprico A;Silva TC;Olsen C;Garofano L;Cava C;Garolini D;Sabedot TS;Malta TM;Pagnotta SM;Castiglioni I;Ceccarelli M;Bontempi G;Noushmehr H
通讯作者：
Noushmehr H
影响因子：
5.3
作者：
Bader, AG;Vogt, PK
通讯作者：
Vogt, PK
影响因子：
5.3
作者：
Evdokimova, V;Ruzanov, P;Sorensen, PHB
通讯作者：
Sorensen, PHB