Department of Gastrointestinal Gland Surgery, the First Affiliated Hospital of Guangxi Medical University, Guangxi Medical University, Nanning, Guangxi, China.
Guangxi Key Laboratory of Enhanced Recovery after Surgery for Gastrointestinal Cancer, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi, China.
Bioengineered. 2022 May;13(5):13970-13985. doi: 10.1080/21655979.2022.2084235.
DEAD-box helicase (DDX) family exerts a critical effect on cancer initiation and progression through alternative splicing, transcription and ribosome biogenesis. Increasing evidence has demonstrated that DEAD-box helicase 56 (DDX56) is over-expressed in several cancers, which plays an oncogenic role. Till the present, the impact of DDX56 on gastric cancer (GC) remains unclear. We conducted high-throughput sequencing (RNA-seq) to demonstrate aberrant DDX56 levels within 10 GC and matched non-carcinoma tissue samples. DDX56 levels were detected through qRT-PCR, western blotting (WB) and immunochemical staining in GC patients. We conducted gain- and loss-of-function studies to examine DDX56's biological role in GC development. In vitro, we carried out 5‑Ethynyl‑2‑deoxyuridine (EdU), scratch, Transwell, and flow cytometry (FCM) assays for detecting GC cell growth, invasion, migration and apoptosis. Additionally, gene set enrichment analysis (GSEA), WB assay, and Encyclopedia of RNA Interactomes (ENCORI) were carried out for analyzing DDX56-regulated downstream genes and signaling pathways. In vivo, tumor xenograft experiment was performed for investigating how DDX56 affected GC development within BALB/c nude mice. Functionally, DDX56 knockdown arrested cell cycle at G1 phase, invasion and migration of AGS and MKN28 cells, and enhanced their apoptosis. Ectopic DDX56 expression enhanced the cell growth, migration and invasion, and inhibited apoptosis. Knockdown of DDX56 suppressed GC growth in the tumor models of BALB/c nude mice. Mechanistically, DDX56 post-transcriptionally suppressed FOXO1/p21 Cip1 protein expression, which could activate its downstream cyclin E1/CDK2/c-Myc signaling pathways. This sheds lights on the GC pathogenic mechanism and offers a potential anti-cancer therapeutic target.
DEAD-box 解旋酶 (DDX) 家族通过选择性剪接、转录和核糖体生物发生对癌症的发生和发展发挥着关键作用。越来越多的证据表明,DEAD-box 解旋酶 56 (DDX56) 在几种癌症中过表达,发挥致癌作用。直到现在,DDX56 对胃癌 (GC) 的影响仍不清楚。我们进行了高通量测序 (RNA-seq),以证明 10 个 GC 和匹配的非癌组织样本中存在异常的 DDX56 水平。通过 qRT-PCR、western blot (WB) 和免疫化学染色检测 GC 患者的 DDX56 水平。我们进行了增益和缺失功能研究,以研究 DDX56 在 GC 发展中的生物学作用。在体外,我们进行了 5-乙炔基-2-脱氧尿苷 (EdU)、划痕、Transwell 和流式细胞术 (FCM) 测定,以检测 GC 细胞的生长、侵袭、迁移和凋亡。此外,进行了基因集富集分析 (GSEA)、WB 测定和 RNA 相互作用组百科全书 (ENCORI),以分析 DDX56 调节的下游基因和信号通路。在体内,进行了肿瘤异种移植实验,以研究 DDX56 如何影响 BALB/c 裸鼠中的 GC 发展。功能上,DDX56 敲低使 AGS 和 MKN28 细胞的细胞周期停滞在 G1 期,侵袭和迁移,并增强其凋亡。DDX56 的异位表达增强了细胞的生长、迁移和侵袭,并抑制了凋亡。DDX56 的敲低抑制了 BALB/c 裸鼠肿瘤模型中的 GC 生长。机制上,DDX56 在后转录水平上抑制 FOXO1/p21 Cip1 蛋白表达,从而激活其下游细胞周期蛋白 E1/CDK2/c-Myc 信号通路。这揭示了 GC 的发病机制,并为潜在的抗癌治疗靶点提供了依据。