Xing Shujian, Zhang Yaqi, Zhang Jing
Department of Gynaecology and Obstetrics, Zouping People's Hospital, Zouping, Shandong 256200, People's Republic of China.
Department of Gynaecology and Obstetrics, Weifang Yidu Central Hospital, Weifang, Shandong 262500, People's Republic of China.
Onco Targets Ther. 2020 Jun 16;13:5643-5655. doi: 10.2147/OTT.S254662. eCollection 2020.
Long intergenic non-protein coding RNA 1224 (LINC01224) plays vital roles in the tumorigenesis and progression of hepatocellular carcinoma. Here, we determined LINC01224 expression in epithelial ovarian cancer (EOC) tissues and cells. We also assessed the effects of LINC01224 knockdown on the malignant phenotype of EOC cells both in vitro and in vivo. Furthermore, the detailed molecular mechanisms underlying the oncogenic actions of LINC01224 in EOC cells were elucidated.
Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect LINC01224 expression in EOC tissues and cells. EOC cells were transfected with small interfering RNAs, and cell proliferation, apoptosis, migration, and invasion were assessed using Cell Counting Kit-8 assay, flow cytometry, cell migration assays, and cell invasion assays, respectively. Using tumor xenografts, the effects of LINC01224 silencing on EOC tumor growth were analyzed in vivo. The mechanism underlying LINC01224 regulation of malignant processes in EOC cells was explored using bioinformatics, RNA immunoprecipitation assay, qRT-PCR, Western blotting, and rescue experiments.
LINC01224 expression was upregulated in EOC tissues and cells. LINC01224 upregulation was correlated to tumor size, the International Federation of Gynecology and Obstetrics stage, and lymph node metastasis. LINC01224 depletion in EOC cells suppressed cell proliferation, migration, and invasion and facilitated cell apoptosis in vitro. LINC01224 downregulation also hindered EOC tumor growth in vivo. Mechanistically, LINC01224 served as a competing endogenous RNA for microRNA-485-5p (miR-485-5p) and consequently increased p21-activated kinase 4 (PAK4) expression in EOC cells. Furthermore, miR-485-5p inhibition or PAK4 upregulation significantly abrogated the effects of LINC01224 depletion in EOC cells.
LINC01224/miR-485-5p/PAK4 formed a competing endogenous RNA network regulating the aggressive behavior of EOC. Therefore, targeting this pathway may be an attractive therapeutic strategy for EOC.
长链基因间非编码RNA 1224（LINC01224）在肝细胞癌的肿瘤发生和进展中起着至关重要的作用。在此，我们测定了LINC01224在上皮性卵巢癌（EOC）组织和细胞中的表达。我们还评估了敲低LINC01224对EOC细胞体外和体内恶性表型的影响。此外，还阐明了LINC01224在EOC细胞中致癌作用的详细分子机制。
采用定量实时聚合酶链反应（qRT-PCR）检测EOC组织和细胞中LINC01224的表达。用小干扰RNA转染EOC细胞，分别使用细胞计数试剂盒-8法、流式细胞术、细胞迁移试验和细胞侵袭试验评估细胞增殖、凋亡、迁移和侵袭。利用肿瘤异种移植模型，在体内分析LINC01224沉默对EOC肿瘤生长的影响。利用生物信息学、RNA免疫沉淀试验、qRT-PCR、蛋白质免疫印迹法和挽救实验探索LINC01224调控EOC细胞恶性进程的机制。
LINC01224在EOC组织和细胞中表达上调。LINC01224上调与肿瘤大小、国际妇产科联盟分期和淋巴结转移相关。EOC细胞中LINC01224的缺失抑制了细胞增殖、迁移和侵袭，并促进了体外细胞凋亡。LINC01224下调也阻碍了EOC肿瘤在体内的生长。机制上，LINC01224作为微小RNA-485-5p（miR-485-5p）的竞争性内源性RNA，从而增加了EOC细胞中p21激活激酶4（PAK4）的表达。此外，miR-485-5p抑制或PAK4上调显著消除了LINC01224缺失对EOC细胞的影响。
LINC01224/miR-485-5p/PAK4形成了一个竞争性内源性RNA网络，调节EOC的侵袭行为。因此，靶向该通路可能是一种有吸引力的EOC治疗策略。
