Department of General Surgery, Shengjing Hospital China Medical University, No. 36 Sanhao Street, Heping District, Shenyang, 110004, China.
Department of Clinical Pharmacy, Shenyang Pharmaceutical University, Shenyang, 110016, China.
BMC Cancer. 2021 Jan 4;21(1):2. doi: 10.1186/s12885-020-07670-y.
MicroRNA (miR)-200c has been widely reported to be involved in colon cancer progress. However, the mechanisms of miR-200c in regulating tumor metastasis and growth remain to be fully elucidated. This study aimed to investigate the mechanism of miR-200c targets fucosyltransferase 4 (FUT4) on the proliferation of colon cancer.
The miR-200c and FUT4 mRNA levels in LoVo and SW480 cells were measured by real-time quantitative polymerase chain reaction. Further, miR-200c mimic, FUT4 siRNA and FUT4 mimic were transfected into cells, separately. Cell counting kit-8, plate colony formation and transwell assays were used to analyse the cells biological behaviour.. Immunofluorescence was used to analyse the Ki-67 expression Moreover, the Wnt/β-catenin pathway-related proteins were detected by western blots. A double luciferase experiment was performed to confirm the relationship between miR-200c and FUT4. In vivo, tumour growth and Wnt/β-catenin pathway-related proteins were also analysed.
In vitro, the expression of miR-200c and FUT4 were negatively correlated in LoVo and SW480 cells (correlation coefficients were - 0.9046 and - 0.9236, respectively). MiR-200c overexpression inhibited the proliferation, migration and invasion of LoVo and SW480 cells by downregulating FUT4. The Ki67-positive cells and Wnt/β-catenin signalling pathway-related proteins were reduced in the miR-200c overexpression and FUT4 silencing groups. A dual luciferase reporting system identified FUT4 as the target of miR-200c. The results in vivo were further confirmed the foundation of cells study.
In summary, miR-200c overexpression inhibits proliferation of colon cancer targeting FUT4 to downregulate the Wnt/β-catenin pathway, which promises molecular targets to inhibit metastasis for colon cancer therapy.
MicroRNA (miR)-200c 已被广泛报道参与结肠癌的进展。然而，miR-200c 调节肿瘤转移和生长的机制仍有待充分阐明。本研究旨在探讨 miR-200c 靶基因岩藻糖基转移酶 4 (FUT4) 对结肠癌增殖的作用机制。
实时定量聚合酶链反应检测 LoVo 和 SW480 细胞中 miR-200c 和 FUT4 mRNA 水平。进一步，分别转染 miR-200c 模拟物、FUT4 siRNA 和 FUT4 模拟物。细胞计数试剂盒-8、平板集落形成和 Transwell 分析用于分析细胞的生物学行为。免疫荧光法分析 Ki-67 的表达。此外，通过 Western blot 检测 Wnt/β-catenin 通路相关蛋白。双荧光素酶实验证实 miR-200c 与 FUT4 的关系。体内，还分析了肿瘤生长和 Wnt/β-catenin 通路相关蛋白。
体外，LoVo 和 SW480 细胞中 miR-200c 和 FUT4 的表达呈负相关（相关系数分别为-0.9046 和-0.9236）。miR-200c 过表达通过下调 FUT4 抑制 LoVo 和 SW480 细胞的增殖、迁移和侵袭。Ki67 阳性细胞和 Wnt/β-catenin 信号通路相关蛋白在 miR-200c 过表达和 FUT4 沉默组中减少。双荧光素酶报告系统鉴定 FUT4 为 miR-200c 的靶基因。体内的结果进一步证实了细胞研究的基础。
综上所述，miR-200c 过表达通过靶向 FUT4 抑制结肠癌增殖，下调 Wnt/β-catenin 通路，为结肠癌治疗提供了抑制转移的分子靶点。
