Graduate School of Dalian Medical University, Dalian Medical University, Dalian, Liaoning, China.
Department of Hepatobiliary Surgery, The Affiliated Changzhou No. 2 People's Hospital of Nanjing Medical University, Changzhou, Jiangsu, China.
Bioengineered. 2022 Apr;13(4):10617-10628. doi: 10.1080/21655979.2022.2064206.
Pancreatic cancer (PC) is a life-threatening cancer with increasing incidence in developed countries. Reports indicate that tRNA-derived fragments (tRFs) are possible therapeutic targets and biomarkers for cancer treatment. Nonetheless, the effect of tRF-Leu-AAG on PC is unclear. This study aims to explore the role of tRF-Leu-AAG and upstream frameshift mutant 1 (UPF1) in the development of PC and its potential underlying mechanisms. High-throughput second-generation sequencing techniques were used to detect the expression of tRFs in cancerous and adjacent normal tissues from PC patients. The role of tRF-Leu-AAG proliferation in PC cells was investigated via the Cell Counting Kit-8 (CCK8) assay. The effect of tRF-Leu-AAG on the invasion and migration ability of PC cells was also determined by the transwell assay. Thereafter, the downstream target genes of tRF-Leu-AAG were comprehensively predicted using bioinformatics analysis databases. We also used the Dual-Luciferase Reporter assay to assess the nexus between tRF-Leu-AAG and UPF1. Eventually, Western Blot was used to validate the expression of UPF1 in PC cells. A total of 33 tRF expressions significantly varied from PC patients. RT-qPCR confirmed that the expression of tRF-Leu-AAG was observably up-regulated in PC cells as compared to the control cells. Importantly, knockdown of tRF-Leu-AAG observably inhibited cell proliferation, migration, and invasion. Furthermore, according to the predicted frameshift database results, the UPF1 acted as downstream target genes for tRF-Leu-AAG and significantly down-regulated UPF1 expression.
胰腺癌（PC）是一种危及生命的癌症，在发达国家的发病率不断上升。有报道称，tRNA 衍生片段（tRFs）可能是癌症治疗的治疗靶点和生物标志物。然而，tRF-Leu-AAG 对 PC 的影响尚不清楚。本研究旨在探讨 tRF-Leu-AAG 和上游移码突变 1（UPF1）在 PC 发生发展中的作用及其潜在机制。采用高通量第二代测序技术检测 PC 患者癌组织和相邻正常组织中 tRFs 的表达。通过细胞计数试剂盒-8（CCK8）检测 tRF-Leu-AAG 对 PC 细胞增殖的作用。通过 Transwell 测定法测定 tRF-Leu-AAG 对 PC 细胞侵袭和迁移能力的影响。此后，利用生物信息学分析数据库综合预测 tRF-Leu-AAG 的下游靶基因。我们还使用双荧光素酶报告基因检测评估 tRF-Leu-AAG 与 UPF1 之间的联系。最后，采用 Western blot 验证 PC 细胞中 UPF1 的表达。共有 33 个 tRF 表达水平在 PC 患者中明显不同。实时荧光定量 PCR 结果证实，与对照组细胞相比，PC 细胞中 tRF-Leu-AAG 的表达明显上调。重要的是，敲低 tRF-Leu-AAG 可明显抑制细胞增殖、迁移和侵袭。此外，根据预测的移码数据库结果，UPF1 作为 tRF-Leu-AAG 的下游靶基因，并显著下调 UPF1 的表达。
