Key Laboratory of Surgery, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China.
Department of Surgery, The First Affiliated Hospital, Wenzhou Medical University, Wenzhou, Zhejiang, China.
J Cell Physiol. 2018 May;233(5):4044-4055. doi: 10.1002/jcp.26072. Epub 2017 Nov 30.
The identification and characterization of long non-coding RNAs (lncRNAs) in diverse biological process has currently developed rapidly. LncRNA-PVT1, located adjacent to the MYC locus on chromosomal region 8q24, has been reported to be associated with many biological processes. However, the function and mechanism of PVT1 in pancreatic carcinoma (PC) is poorly understood. In this present study, we first measured the level of PVT1 in the PC cell lines and tissues by quantitative real-time PCR (qRT-PCR), and then employed loss-of-function and gain-of-function approaches to explore the association between PVT1 expression levels and PC cell proliferation/migration ability. Furthermore, bioinformatics analysis was utilized to show that PVT1 contains binding site for miR-448 and an inverse correlation between PVT1 and miR-448 was obtained in PC specimens. Additionally, dual luciferase reporter assay, RNA-binding protein immunoprecipitation (RIP) and applied biotin-avidin pulldown system were applied to further confirm that PVT1 directly bind with microRNA binding site harboring in the PVT1 sequence. Then, SERBP1 was identified as a target of miR-448 according to the gene expression array analysis of PC clinical samples. Together, we revealed that PVT1 functions as an endogenous "sponge" by competing for miR-448 binding to regulate the miRNA target SERBP1 and, therefore, promotes the proliferation and migration of PC cells.
长链非编码 RNA（lncRNA）在各种生物过程中的鉴定和特征目前发展迅速。位于染色体 8q24 区域上 MYC 基因座附近的 lncRNA-PVT1 已被报道与许多生物过程有关。然而，PVT1 在胰腺癌（PC）中的功能和机制仍知之甚少。在本研究中，我们首先通过实时定量 PCR（qRT-PCR）测量了 PC 细胞系和组织中 PVT1 的水平，然后采用功能丧失和功能获得方法来探讨 PVT1 表达水平与 PC 细胞增殖/迁移能力之间的关系。此外，生物信息学分析表明 PVT1 包含 miR-448 的结合位点，并且在 PC 标本中获得了 PVT1 和 miR-448 之间的负相关。此外，双荧光素酶报告基因检测、RNA 结合蛋白免疫沉淀（RIP）和应用生物素-亲和素下拉系统进一步证实 PVT1 直接与 PVT1 序列中包含的 microRNA 结合位点结合。然后，根据 PC 临床样本的基因表达谱分析，鉴定出 SERBP1 是 miR-448 的靶基因。综上所述，我们揭示了 PVT1 通过竞争 miR-448 的结合作为内源性“海绵”发挥作用，从而调节 miRNA 靶标 SERBP1，进而促进 PC 细胞的增殖和迁移。
