Doctoral Program in Methods and Applications in Life Sciences, Faculty of Biology, Campus Lagoas-Marcosende, Universidade de Vigo, 36310 Vigo, Spain.
Liquid Biopsy Analysis Unit, Translational Medical Oncology (Oncomet), Health Research Institute of Santiago de Compostela (IDIS), CIBERONC, Travesía da Choupana, 15706 Santiago de Compostela, Spain.
Int J Mol Sci. 2022 Jul 30;23(15):8463. doi: 10.3390/ijms23158463.
The present study explored the impact of inhibiting α(1,6)fucosylation (core fucosylation) on the functional phenotype of a cellular model of colorectal cancer (CRC) malignization formed by the syngeneic SW480 and SW620 CRC lines. Expression of the gene encoding α(1,6)fucosyltransferase was inhibited in tumor line SW480 by a combination of shRNA-based antisense knockdown and agglutinin (LCA) selection. LCA-resistant clones were subsequently assayed in vitro for proliferation, migration, and adhesion. The α(1,6)FT-inhibited SW480 cells showed enhanced proliferation in adherent conditions, unlike their α(1,6)FT-depleted SW620 counterparts, which displayed reduced proliferation. Under non-adherent conditions, α(1,6)FT-inhibited SW480 cells also showed greater growth capacity than their respective non-targeted control (NTC) cells. However, cell migration decreased in SW480 after knockdown, while adhesion to EA.hy926 cells was significantly enhanced. The reported results indicate that the knockdown strategy with subsequent selection for LCA-resistant clones was effective in greatly reducing α(1,6)FT expression in SW480 and SW620 CRC lines. In addition, α(1,6)FT impairment affected the proliferation, migration, and adhesion of α(1,6)FT-deficient clones SW480 and SW620 in a tumor stage-dependent manner, suggesting that core fucosylation has a dynamic role in the evolution of CRC.
本研究探讨了抑制 α(1,6)岩藻糖基转移酶（核心岩藻糖基化）对结直肠癌（CRC）恶性转化的细胞模型功能表型的影响，该模型由同源 SW480 和 SW620 CRC 细胞系形成。通过 shRNA 反义敲低和凝集素（LCA）选择的组合，抑制肿瘤系 SW480 中编码 α(1,6)岩藻糖基转移酶的 基因的表达。随后，在体外检测 LCA 抗性克隆的增殖、迁移和粘附。与耗尽 α(1,6)FT 的 SW620 对照相比，α(1,6)FT 抑制的 SW480 细胞在贴壁条件下表现出增强的增殖，而不是增殖减少。在非贴壁条件下，α(1,6)FT 抑制的 SW480 细胞的生长能力也强于各自的非靶向对照（NTC）细胞。然而，在 knockdown 后，SW480 中的细胞迁移减少，而与 EA.hy926 细胞的粘附显著增强。报道的结果表明，用 LCA 抗性克隆进行后续选择的 knockdown 策略在很大程度上降低了 SW480 和 SW620 CRC 细胞系中 α(1,6)FT 的表达。此外，α(1,6)FT 损伤以肿瘤阶段依赖的方式影响 α(1,6)FT 缺陷克隆 SW480 和 SW620 的增殖、迁移和粘附，表明核心岩藻糖基化在 CRC 的进化中具有动态作用。
