Department of Oncology, Ruijin Hospital, Shanghai Jiaotong University School of Medicine , Shanghai, China.
State Key Laboratory for Medical Genomics, Shanghai Institute of Hematology and Collaborative Innovation Center of Hematology, National Research Center for Translational Medicine, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine and School of Life Sciences and Biotechnology , Shanghai, China.
Cell Cycle. 2020 Nov;19(21):2939-2954. doi: 10.1080/15384101.2020.1829824. Epub 2020 Oct 12.
MicroRNA functions as an oncogenic regulator or tumor suppressor in various human tumors. Although bioinformatics analysis suggested that miRNA-20b-5p may be associated with the tumorigenesis, its role in colon cancer remains elusive. To investigate the role of miRNA-20b-5p, HCT116 cell, a human colon cancer cell line used in therapeutic research and drug screenings, was chosen as a model system for our in vitro studies. We first carried out bioinformatics and microarray analysis. To gain further mechanism insight, flow cytometry was performed to determine cell apoptosis and cell cycle, and western blot or immunohistochemistry were employed to check the expression of CCND1/CDK/FOXM1 axis in HCT116 cells. In addition, wound-healing migration assay and transwell assay were conducted to uncover the effect of miR-20b-5p on tumor migration and invasion. Finally, we examined the role of miR-20b-5p by subcutaneous xenograft mouse models. Our data have shown that miRNA-20b-5p inhibited the cell cycle, migration, and invasion in HCT116 cells, but had no effect on cell apoptosis. CyclinD1 (CCND1) was identified as a direct target of miR-20b-5p. Overexpression of miRNA-20b-5p downregulated level in HCT-116 cells. Mechanically, the inhibition of cell cycle, migration, and invasion of CC cells mediated by miRNA-20b-5p are through regulating the CCND1/CDK4/FOXM1 axis. Furthermore, miRNA-20b-5p inhibited the tumorigenesis in Balb/c nude mice CC xenograft models. Our data demonstrated that miR-20b-5p may serve as a tumor suppressor in colon cancer by negatively regulating , implying that miR-20b-5p could be a potential therapeutic target for the treatment of colon cancer.
microRNA 在各种人类肿瘤中作为致癌调节剂或肿瘤抑制因子发挥作用。尽管生物信息学分析表明 miRNA-20b-5p 可能与肿瘤发生有关，但它在结肠癌中的作用仍不清楚。为了研究 miRNA-20b-5p 的作用，选择 HCT116 细胞（一种用于治疗研究和药物筛选的人结肠癌细胞系）作为我们体外研究的模型系统。我们首先进行了生物信息学和微阵列分析。为了获得进一步的机制见解，我们进行了流式细胞术来确定细胞凋亡和细胞周期，并用 Western blot 或免疫组化来检查 HCT116 细胞中 CCND1/CDK/FOXM1 轴的表达。此外，进行了划痕愈合迁移实验和 Transwell 实验以揭示 miR-20b-5p 对肿瘤迁移和侵袭的影响。最后，我们通过皮下异种移植小鼠模型检验了 miR-20b-5p 的作用。我们的数据表明，miRNA-20b-5p 抑制了 HCT116 细胞的细胞周期、迁移和侵袭，但对细胞凋亡没有影响。细胞周期蛋白 D1（CCND1）被鉴定为 miR-20b-5p 的直接靶标。miR-20b-5p 的过表达下调了 HCT-116 细胞中的水平。机制上，miRNA-20b-5p 通过调节 CCND1/CDK4/FOXM1 轴抑制 CC 细胞的细胞周期、迁移和侵袭。此外，miRNA-20b-5p 抑制了 Balb/c 裸鼠 CC 异种移植模型中的肿瘤发生。我们的数据表明，miR-20b-5p 可能通过负调控在结肠癌中作为肿瘤抑制因子发挥作用，暗示 miR-20b-5p 可能成为结肠癌治疗的潜在治疗靶点。
