| 王 硕,孔睿佼,井 杰,刘海东,贾 音,刘善荣.Circ-CL5A1受TDP-43调控并抑制肝细胞癌转移的研究[J].,2020,(11):2036-2041 |
| Circ-CL5A1受TDP-43调控并抑制肝细胞癌转移的研究 |
| Circ-COL5A1 is Regulated by TDP-43 and Inhibit HCC Metastasis |
| 投稿时间：2020-02-23 修订日期：2020-03-18 |
| DOI：10.13241/j.cnki.pmb.2020.11.007 |
| 中文关键词: 肝细胞癌；环状RNA；RNA结合蛋白；Circ-COL5A1 TDP-43 |
| 英文关键词: Hepatocellular carcinoma Circular RNA RNA binding protein Circ-COL5A1 TDP-43 |
| 基金项目:国家自然科学基金重点项目（81730076） |
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| 中文摘要: |
| 摘要 目的：探索Circ-COL5A1的生物学功能、调控机制和作用机制，进而为HCC转移的干预提供候选分子并进一步了解HCC转移。方法：通过前期工作基础选定目标分子Circ-COL5A1。通过慢病毒转染在HCC细胞系中过表达Circ-COL5A1，进而通过划痕愈合实验、transwell实验观察Circ-COL5A1的生物学功能。通过生物信息学分析、表达干扰实验和RNA免疫共沉淀（RIP）实验探究目标分子的调控机制。通过western blot 技术、实时定量PCR（qRT-PCR）技术对目标分子的下游作用机制进行初步探索。结果：Circ-COL5A1在肝癌干细胞中表达下调，而且Circ-COL5A1过表达的HCC细胞系侵袭和迁移能力减弱。在Circ-COL5A1生物学合成过程中，RNA结合蛋白TDP-43可以富集其线性前体，并在环化结构形成后解离。Circ-COL5A1还可以降低其亲本基因V型胶原蛋白α1链(COL5A1)的蛋白质表达水平，这可能会影响多个信号通路进而干预HCC的转移过程。结论：内源性的Circ-COL5A1可以抑制HCC的转移能力，可以为阻断HCC转移提供候选分子。TDP-43的促进环状RNA形成提示RNA结合蛋白是环状RNA生物学合成过程中的重要调控因子。Circ-COL5A1可以通过转录后调控抑制其亲本基因COL5A1的表达。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the biological function, mechanism and regulation factor of circ-COL5A1, providing candidate molecules for the intervention and further understanding of HCC metastasis. Methods: The target molecule Circ-COL5A1 were selected based on the preliminary work. The biological function of circ-COL5A1 is verified through transwell assay and wound healing assay. Bioinformatic analysis, gene expression interference and RNA immunoprecipitation (RIP) experiments were employed to explore the regulation mechanism of circ-COL5A1. Western blot and quantitative real-time PCR (qRT-PCR) were employed to explore the downstream pathway of circ-COL5A1. Results: The expression of Circ-COL5A1 is down-regulated in HCC cancer stem cells. And Circ-COL5A1 overexpressed inhibit the invasion and migration ability in Huh7 and HCC-LM3 cell lines. The result of RIP experiments showed that RNA-binding protein (RBP) TDP-43 can enrich the linear precursor and dissociate after the formation of the circular structure. Circ-COL5A1 can also reduce the protein expression of its maternal gene COL51, which may affect multiple signaling pathways and thus interfere with the metastasis process of HCC. Conclusion: Endogenous Circ-COL5A1 can inhibit the metastasis ability of HCC cell lines, and may provide candidate molecules for blocking HCC metastasis. TDP-43 promotes the biogenesis of Circ-COL5A1, suggesting that RBPs are important regulators in the process of circRNA biogenesis. Circ-COL5A1 inhibits protein expression of its maternal genes COL5A1 through post-transcription regulation. |
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