长链非编码 RNA MILIP 将 YBX1 与 Snai1 的翻译激活联系起来，并促进肾透明细胞癌的转移。
LncRNA MILIP links YBX1 to translational activation of Snai1 and promotes metastasis in clear cell renal cell carcinoma.
机构信息
Department of Nephrology, Henan Provincial Key Laboratory of Kidney Disease and Immunology, Henan Provincial Clinical Research Center for Kidney Disease, Henan Provincial People's Hospital and People's Hospital of Zhengzhou University, Zhengzhou, China.
School of Medicine and Public Health, The University of Newcastle, Newcastle, Australia.
出版信息
J Exp Clin Cancer Res. 2022 Aug 26;41(1):260. doi: 10.1186/s13046-022-02452-9.
BACKGROUND
Distant metastasis is the major cause of clear cell renal cell carcinoma (ccRCC)-associated mortality. However, molecular mechanisms involved in ccRCC metastasis remain to be fully understood. With the increasing appreciation of the role of long non-coding RNAs (lncRNAs) in cancer development, progression, and treatment resistance, the list of aberrantly expressed lncRNAs contributing to ccRCC pathogenesis is expanding rapidly.
METHODS
Bioinformatics analysis was carried out to interrogate publicly available ccRCC datasets. In situ hybridization and qRT-PCR assays were used to test lncRNA expression in human ccRCC tissues and cell lines, respectively. Chromatin immunoprecipitation and luciferase reporter assays were used to examine transcriptional regulation of gene expression. Wound healing as well as transwell migration and invasion assays were employed to monitor ccRCC cell migration and invasion in vitro. ccRCC metastasis was also examined using mouse models in vivo. RNA pulldown and RNA immunoprecipitation were performed to test RNA-protein associations, whereas RNA-RNA interactions were tested using domain-specific chromatin isolation by RNA purification.
RESULTS
MILIP expression was upregulated in metastatic compared with primary ccRCC tissues. The increased MILIP expression in metastatic ccRCC cells was driven by the transcription factor AP-2 gamma (TFAP2C). Knockdown of MILIP diminished the potential of ccRCC cell migration and invasion in vitro and reduced the formation of ccRCC metastatic lesions in vivo. The effect of MILIP on ccRCC cells was associated with alterations in the expression of epithelial-to-mesenchymal transition (EMT) hallmark genes. Mechanistically, MILIP formed an RNA-RNA duplex with the snail family transcriptional repressor 1 (Snai1) mRNA and bound to Y-box binding protein 1 (YBX1). This promoted the association between the YBX1 protein and the Snai1 mRNA, leading to increased translation of the latter. Snai1 in turn played an important role in MILIP-driven ccRCC metastasis.
CONCLUSIONS
The TFAP2C-responsive lncRNA MILIP drives ccRCC metastasis. Targeting MILIP may thus represent a potential avenue for ccRCC treatment.
背景
远处转移是导致透明细胞肾细胞癌（ccRCC）相关死亡的主要原因。然而，ccRCC 转移涉及的分子机制仍有待充分了解。随着人们对长链非编码 RNA（lncRNA）在癌症发生、发展和治疗耐药中的作用认识的不断提高，导致 ccRCC 发病机制的异常表达 lncRNA 清单迅速扩大。
方法
通过生物信息学分析，对公开的 ccRCC 数据集进行了查询。原位杂交和 qRT-PCR 检测分别用于检测人 ccRCC 组织和细胞系中的 lncRNA 表达。染色质免疫沉淀和荧光素酶报告基因检测用于检测基因表达的转录调控。体外伤口愈合以及 Transwell 迁移和侵袭实验用于监测 ccRCC 细胞的迁移和侵袭。体内也使用小鼠模型来检测 ccRCC 的转移。RNA 下拉和 RNA 免疫沉淀用于检测 RNA-蛋白的相互作用，而使用特定于结构域的 RNA 纯化进行 RNA-RNA 相互作用检测。
结果
MILIP 在转移性 ccRCC 组织中表达上调，与原发性 ccRCC 组织相比。在转移性 ccRCC 细胞中，MILIP 的表达增加是由转录因子 AP-2γ（TFAP2C）驱动的。MILIP 的敲低减少了 ccRCC 细胞体外迁移和侵袭的潜力，并减少了体内 ccRCC 转移病变的形成。MILIP 对 ccRCC 细胞的作用与上皮-间充质转化（EMT）特征基因表达的改变有关。从机制上讲，MILIP 与 snail 家族转录抑制因子 1（Snai1）mRNA 形成 RNA-RNA 双链，并与 Y 盒结合蛋白 1（YBX1）结合。这促进了 YBX1 蛋白与 Snai1 mRNA 的结合，导致后者的翻译增加。Snai1 反过来在 MILIP 驱动的 ccRCC 转移中发挥了重要作用。
结论
TFAP2C 反应性 lncRNA MILIP 驱动 ccRCC 转移。因此，靶向 MILIP 可能是 ccRCC 治疗的一种潜在途径。
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