Zhengzhou University, No. 100 Science Ave, Gaoxin District, Zhengzhou 450001, China.
Medical Department, Affiliated Hospital of Shandong University of Traditional Chinese Medicine, No. 16369 Jingshi Road, Jinan 250011, China.
J Biochem. 2020 Nov 1;168(5):535-546. doi: 10.1093/jb/mvaa073.
Long non-coding RNA (lnc) HCG18 has been reported to contribute progression of a variety of tumours. However, its roles in hepatocellular carcinoma (HCC) remains unknown. In the current study, we intended to uncover the biological functions of HCG18 in HCC. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to detect the expression of HCG18, microRNA-214-3p (miR-214-3p) and centromere protein M (CENPM) messenger RNA (mRNA). The role of HCG18 in the growth and migration were assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, colony formation assay, wound healing assay and flow cytometry in vitro and animal experiments in vivo. The results showed that HCG18 was highly expressed in HCC tissues. HCG18 silencing inhibited the proliferation and migration while induced the apoptosis of HCC cells. Besides, miR-214-3p was down-regulated in HCC cells. Further experiments revealed that miR-214-3p could directly bind to HCG18 and exerted an anti-tumour role to counteracted siHCG18-1-mediated influence in HCC cells. Moreover, miR-214-3p could directly interact with CENPM mRNA and down-regulating the expression of CENPM. While HCG18 could up-regulate the expression of CENPM through acting as a sponge of miR-214-3p. Therefore, those results suggested HCG18 functioned as an oncogene to promote the proliferation and migration of HCC cells via miR-214-3p/CENPM axis.
长链非编码 RNA (lnc)HCG18 已被报道有助于多种肿瘤的进展。然而,其在肝细胞癌 (HCC) 中的作用尚不清楚。在本研究中,我们旨在揭示 HCG18 在 HCC 中的生物学功能。采用实时定量聚合酶链反应 (qRT-PCR) 检测 HCG18、微小 RNA-214-3p (miR-214-3p) 和着丝粒蛋白 M (CENPM) 信使 RNA (mRNA) 的表达。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法、集落形成实验、划痕愈合实验和流式细胞术体外以及动物实验体内评估 HCG18 在 HCC 细胞生长和迁移中的作用。结果显示,HCG18 在 HCC 组织中高表达。HCG18 沉默抑制 HCC 细胞的增殖和迁移,同时诱导细胞凋亡。此外,miR-214-3p 在 HCC 细胞中下调。进一步的实验表明,miR-214-3p 可以直接与 HCG18 结合,发挥抑癌作用,抵消 siHCG18-1 介导的 HCC 细胞的影响。此外,miR-214-3p 可以直接与 CENPM mRNA 相互作用并下调 CENPM 的表达。而 HCG18 可以通过作为 miR-214-3p 的海绵体来上调 CENPM 的表达。因此,这些结果表明 HCG18 通过 miR-214-3p/CENPM 轴作为癌基因促进 HCC 细胞的增殖和迁移。