Cancer Inflammation Laboratory, Olivia Newton-John Cancer Research Institute, Heidelberg, Victoria, Australia.
School of Cancer Medicine, La Trobe University, Bundoora, Victoria, Australia.
Proteomics. 2021 Jul;21(13-14):e2000098. doi: 10.1002/pmic.202000098. Epub 2021 May 28.
Doublecortin-like kinase 1 (DCLK1) is a putative cancer stem cell marker, a promising diagnostic and prognostic maker for malignant tumors and a proposed driver gene for gastric cancer (GC). DCLK1 overexpression in a majority of solid cancers correlates with lymph node metastases, advanced disease and overall poor-prognosis. In cancer cells, DCLK1 expression has been shown to promote epithelial-to-mesenchymal transition (EMT), driving disruption of cell-cell adhesion, cell migration and invasion. Here, we report that DCLK1 influences small extracellular vesicle (sEV/exosome) biogenesis in a kinase-dependent manner. sEVs isolated from DCLK1 overexpressing human GC cell line MKN1 (MKN1 -sEVs), promote the migration of parental (non-transfected) MKN1 cells (MKN1 ). Quantitative proteome analysis of MKN1 -sEVs revealed enrichment in migratory and adhesion regulators (STRAP, CORO1B, BCAM, COL3A, CCN1) in comparison to MKN1 -sEVs. Moreover, using DCLK1-IN-1, a specific small molecule inhibitor of DCLK1, we reversed the increase in sEV size and concentration in contrast to other EV subtypes, as well as kinase-dependent cargo selection of proteins involved in EV biogenesis (KTN1, CHMP1A, MYO1G) and migration and adhesion processes (STRAP, CCN1). Our findings highlight a specific role of DCLK1-kinase dependent cargo selection for sEVs and shed new light on its role as a regulator of signaling in gastric tumorigenesis.
双皮质醇激酶 1（DCLK1）是一种假定的癌症干细胞标志物，是恶性肿瘤有前途的诊断和预后标志物，也是胃癌（GC）的候选驱动基因。大多数实体瘤中 DCLK1 的过表达与淋巴结转移、晚期疾病和整体预后不良相关。在癌细胞中，DCLK1 的表达已被证明可促进上皮-间充质转化（EMT），从而破坏细胞-细胞黏附、细胞迁移和侵袭。在这里，我们报告 DCLK1 以激酶依赖的方式影响小细胞外囊泡（sEV/exosome）的生物发生。来自 DCLK1 过表达的人 GC 细胞系 MKN1（MKN1-sEVs）的 sEVs 促进亲本（未转染）MKN1 细胞（MKN1）的迁移。与 MKN1-sEVs 相比，MKN1-sEVs 的定量蛋白质组分析显示迁移和粘附调节剂（STRAP、CORO1B、BCAM、COL3A、CCN1）富集。此外，使用 DCLK1-IN-1，一种 DCLK1 的特异性小分子抑制剂，我们逆转了 sEV 大小和浓度的增加，与其他 EV 亚型相比，以及与 EV 生物发生（KTN1、CHMP1A、MYO1G）和迁移和粘附过程（STRAP、CCN1）相关的蛋白的激酶依赖性货物选择。我们的研究结果强调了 DCLK1-激酶依赖性货物选择对 sEV 的特定作用，并为其作为胃癌发生中信号转导调节剂的作用提供了新的认识。
