Circ-ASH2L promotes tumor progression by sponging miR-34a to regulate Notch1 in pancreatic ductal adenocarcinoma
Circ-ASH2L promotes tumor progression by sponging miR-34a to regulate Notch1 in pancreatic ductal adenocarcinoma
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DOI：
10.1186/s13046-019-1436-0
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发表时间：
2019-11-12
影响因子：
11.3
通讯作者：
Wang, Shuguang
中科院分区：
文献类型：
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作者：
Chen, Yan;Li, Zhonghu;Wang, Shuguang
研究背景近年来发现环状RNA（CircRNA）在多种肿瘤中发挥重要作用。然而，其在胰腺导管腺癌（PDAC）中的详细作用和调节机制尚不清楚。本研究旨在鉴定PDAC细胞和组织中富集的circRNA并检测其功能和机制。方法在前人研究的基础上，采用circRNA芯片技术鉴定circRNA-ASH 2L（circ-ASH 2L），并进一步采用qRT-PCR技术检测PDAC细胞和标本中circ-ASH 2L的表达。通过transwell、EdU、细胞周期或管形成测定来鉴定circ-ASH 2L的功能。通过WB、RIP、FISH、双荧光素酶测定、RNA下拉或其他测定来探索circ-ASH 2L的调控机制。结果在前期研究的基础上，我们鉴定了一种circRNA（circ-ASH 2L），检测了其在不同恶性肿瘤细胞中的表达，发现circ-ASH 2L在胰腺细胞或肿瘤组织中高表达，并与肿瘤恶性程度相关。进一步研究发现，circ-ASH 2L通过调控miR-34 a，进而调控Notch 1的表达，促进肿瘤的侵袭、增殖和血管生成。Circ-ASH 2L充当miR-34 a的miRNA海绵并促进体内肿瘤进展。最后，我们分析了临床组织中circ-ASH 2L的表达，发现circ-ASH 2L的高表达与淋巴浸润和TNM分期相关，并且是胰腺患者生存的独立危险因素。结论circ-ASH 2L在肿瘤侵袭中起重要作用，circ-ASH 2L水平升高可能是PDAC诊断或进展的一个有用指标。
Background Circular RNAs (circRNAs) have recently been shown to play important roles in different tumors. However, their detailed roles and regulatory mechanisms in pancreatic ductal adenocarcinoma (PDAC) are not well understood. This study aimed to identify enriched circRNAs and detect their functions and mechanisms in PDAC cells and tissues. Methods circRNA-ASH2L (circ-ASH2L) was identified by circRNA microarray studies based on previous studies, and further detected in PDAC cells and samples by qRT-PCR. The functions of circ-ASH2L were identified by transwell, EdU, cell cycle or Tube formation assays. The regulatory mechanisms of circ-ASH2L were explored by WB, RIP, FISH, dual-luciferase assays, RNA pulldown or other assays. Results We identified a circRNA (circ-ASH2L) based on our previous studies, detected its expression in different malignant cells and found that circ-ASH2L was highly expressed in pancreatic cells or tumor tissues and correlated with tumor malignancy. Further studies revealed that circ-ASH2L promoted tumor invasion, proliferation and angiogenesis by regulating miR-34a, thus regulate Notch 1 expression. Circ-ASH2L served as a miRNA sponge for miR-34a and promoted tumor progression in vivo. Finally, we analyzed circ-ASH2L expression in clinical tissues and found that high circ-ASH2L expression was correlated with lymphatic invasion and TNM stage and was an independent risk factor for pancreatic patient survival. Conclusions circ-ASH2L play an important role in tumor invasion, and high circ-ASH2L may be a useful marker of PDAC diagnosis or progression.