Department of Clinical Laboratory, Affiliated Hospital of Beihua University, Jilin, China.
Department of Diabetes, Endocrinology and Nutrition, Graduate School of Medicine, Kyoto University, Kyoto, Japan.
Kaohsiung J Med Sci. 2023 Nov;39(11):1062-1076. doi: 10.1002/kjm2.12749. Epub 2023 Sep 12.
To investigate the biological role and mechanism of circ_0084188 in colorectal cancer (CRC). Real-time quantitative polymerase chain reaction and western blot assay were used to detect RNA levels and protein levels in CRC cell lines (HCT116 and SW480), respectively. Cell proliferation was evaluated by Cell Counting Kit-8 assay, 5-ethynyl-2'-deoxyuridine assay, and colony formation assays. Cell apoptosis was determined using flow cytometry. Cell migration and invasion were measured by transwell assay. Sphere formation efficiency was determined by sphere formation assay. The interaction between microRNA-654-3p (miR-654-3p) and circ_0084188 or Kruppel-like factor 12 (KLF12) was confirmed by a dual-luciferase reporter, RNA immunoprecipitation and RNA pull-down assays. Xenograft in CRC mice model was utilized for exploring the role of circ_0084188 in vivo.Circ_0084188 was overexpressed in CRC tissues and cells. Circ_0084188 silencing suppressed cell proliferation, migration, invasion, and stemness and induced apoptosis in CRC cells. Circ_0084188 acted as a sponge for miR-654-3p, and circ_0084188 regulated CRC cell behaviors via sponging miR-654-3p. Moreover, KLF12 was a target of miR-654-3p, and miR-654-3p overexpression inhibited the malignant behaviors of CRC cells by downregulating KLF12. Mechanically, circ_0084188 sponged miR-654-3p to regulate KLF12 expression in CRC cells. In addition, circ_0084188 downregulation inhibited tumor growth in vivo.Circ_0084188 knockdown might repress CRC progression partially via regulating the miR-654-3p/KLF12 axis, providing a novel insight into the pathogenesis of CRC.
为了研究 circ_0084188 在结直肠癌(CRC)中的生物学作用和机制。分别使用实时定量聚合酶链反应和 Western blot 检测 CRC 细胞系(HCT116 和 SW480)中的 RNA 水平和蛋白水平。通过细胞计数试剂盒-8 测定、5-乙炔基-2'-脱氧尿苷测定和集落形成测定评估细胞增殖。通过流式细胞术测定细胞凋亡。通过 Transwell 测定法测定细胞迁移和侵袭。通过球体形成测定法测定球体形成效率。通过双荧光素酶报告、RNA 免疫沉淀和 RNA 下拉测定证实 microRNA-654-3p(miR-654-3p)与 circ_0084188 或 Kruppel 样因子 12(KLF12)之间的相互作用。利用 CRC 小鼠模型进行体内研究探索 circ_0084188 的作用。circ_0084188 在 CRC 组织和细胞中过表达。circ_0084188 沉默抑制 CRC 细胞的增殖、迁移、侵袭和干性,并诱导细胞凋亡。circ_0084188 作为 miR-654-3p 的海绵,通过海绵 miR-654-3p 调节 CRC 细胞行为。此外,KLF12 是 miR-654-3p 的靶标,miR-654-3p 通过下调 KLF12 抑制 CRC 细胞的恶性行为。机械上,circ_0084188 海绵 miR-654-3p 调节 CRC 细胞中的 KLF12 表达。此外,circ_0084188 下调抑制体内肿瘤生长。circ_0084188 敲低可能通过调节 miR-654-3p/KLF12 轴部分抑制 CRC 进展,为 CRC 的发病机制提供了新的见解。