Department of Pathology, The Central Hospital of Wuhan, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, PR China.
Laboratory of Clinical Immunology, Wuhan No.1 Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, PR China.
Life Sci. 2021 Feb 15;267:118928. doi: 10.1016/j.lfs.2020.118928. Epub 2020 Dec 31.
Recently, long noncoding RNAs (lncRNAs) have been reported to play important role in the pathogenesis of various cancers. However, the functions of RNF185-AS1 in hepatocellular carcinoma (HCC) remain unknown.
The RNF185-AS1 expression in HCC cells and tissues was measured by quantitative real-time polymerase chain reaction (qRT-PCR). The effects of RNF185-AS1 on tumor cell proliferation, migration and invasion were assessed by Cell Counting Kit-8 (CCK8) assay, colony formation assay, transwell assay. The luciferase reporter assay, RNA-binding protein immunoprecipitation assay, qRT-PCR and Western blot were performed to explore and confirm the interaction between RNF185-AS1 and miR-221-5p and integrin β5. The role of RNF185-AS1 in tumor progression was explored through in vivo experiments.
RNF185-AS1 was highly expressed in HCC tissues and cell lines. High levels of RNF185-AS1 were correlated with advanced TNM stage, distant metastasis and a poorer overall survival rate. RNF185-AS1 knockdown inhibited cell proliferation, migration and invasion. Additionally, RNF185-AS1 acted as a sponge for miR-221-5p and integrin β5 was identified as a target gene of miR-221-5p. Rescue assays showed that miR-221-5p inhibitor or integrin β5 overexpression rescued the function of RNF185-AS1 knockdown on cell proliferation, migration, and invasion. Moreover, we found that RNF185-AS1 knockdown inhibited tumor growth and metastasis.
Our study demonstrates that RNF185-AS1 is a new oncogenic lncRNA in HCC and suggests that the RNF185-AS1/miR-221-5p/integrin β5 axis might be a potential therapeutic target for HCC.
最近，长链非编码 RNA（lncRNA）被报道在各种癌症的发病机制中发挥重要作用。然而，RNF185-AS1 在肝细胞癌（HCC）中的功能尚不清楚。
通过实时定量聚合酶链反应（qRT-PCR）测量 HCC 细胞和组织中的 RNF185-AS1 表达。通过细胞计数试剂盒-8（CCK8）测定、集落形成测定、transwell 测定评估 RNF185-AS1 对肿瘤细胞增殖、迁移和侵袭的影响。进行荧光素酶报告基因测定、RNA 结合蛋白免疫沉淀测定、qRT-PCR 和 Western blot 以探索和确认 RNF185-AS1 与 miR-221-5p 和整合素β5 之间的相互作用。通过体内实验探索 RNF185-AS1 在肿瘤进展中的作用。
RNF185-AS1 在 HCC 组织和细胞系中高表达。高水平的 RNF185-AS1 与较晚的 TNM 分期、远处转移和较差的总生存率相关。RNF185-AS1 敲低抑制细胞增殖、迁移和侵袭。此外，RNF185-AS1 作为 miR-221-5p 的海绵，并且鉴定出整合素β5 是 miR-221-5p 的靶基因。挽救实验表明，miR-221-5p 抑制剂或整合素β5 过表达挽救了 RNF185-AS1 敲低对细胞增殖、迁移和侵袭的功能。此外，我们发现 RNF185-AS1 敲低抑制肿瘤生长和转移。
本研究表明 RNF185-AS1 是 HCC 中的一种新的致癌 lncRNA，并表明 RNF185-AS1/miR-221-5p/整合素β5 轴可能是 HCC 的潜在治疗靶点。
