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Invest Ophthalmol Vis Sci. 2020 Jun 3;61(6):41. doi: 10.1167/iovs.61.6.41.
The tumor-initiating function of long non-coding RNA (lncRNA), zinc finger protein multitype 2 antisense RNA 1 (ZFPM2-AS1) was reported in lung cancer, yet the relevance of ZFPM2-AS1 in retinoblastoma (RB), a malignancy representing 2.5% to 4% incidence of cancers among children, has not been elucidated. Thus, we attempted to assess the effect of ZFPM2-AS1 and underlying mechanism in RB progression.
First, comparing the differentially expressed lncRNAs in normal retinal tissues as well as RB tissues, the target lncRNA ZFPM2-AS1 was screened out. We then assayed the ZFPM2-AS1 expression in three RB cell lines, and carried out methylthiazol tetrazolium (MTT), transwell assays, and flow cytometric analyses to examine the role of si-ZFPM2-AS1 on cell behaviors. Following online database predication, the correlations between ZFPM2-AS1 and microR-515 (miR-515) or homeobox A1 (HOXA1) were corroborated by dual-luciferase reporter gene assays. Quantitative real-time PCR along with Western blot assays was fulfilled to ascertain the expression of relevant genes.
ZFPM2-AS1 was significantly overexpressed in RB tissues and cell lines, and ZFPM2-AS1 silencing curtailed the growth and metastasis of RB cells both in vitro and in vivo. Bioinformatic websites and dual-luciferase reporter gene assays disclosed that ZFPM2-AS1 might perform as a competing endogenous RNA for miR-515 and positively correlate with HOXA1 to activate the Wnt/β-catenin signaling pathway.
Altogether, these data demonstrated that ZFPM2-AS1 interacted with HOXA1 to promote RB development via mediating miR-515, establishing a promising therapeutic biomarker for RB and prognosis.
长链非编码 RNA（lncRNA）肿瘤起始功能已在肺癌中报道，然而锌指蛋白多型 2 反义 RNA 1（ZFPM2-AS1）在视网膜母细胞瘤（RB）中的相关性尚未阐明，RB 是一种儿童癌症发病率占 2.5%至 4%的恶性肿瘤。因此，我们试图评估 ZFPM2-AS1 在 RB 进展中的作用及其潜在机制。
首先，比较正常视网膜组织和 RB 组织中差异表达的 lncRNA，筛选出靶 lncRNA ZFPM2-AS1。然后在三种 RB 细胞系中检测 ZFPM2-AS1 的表达，并进行甲基噻唑四唑（MTT）、transwell 测定和流式细胞分析，以检查 si-ZFPM2-AS1 对细胞行为的作用。通过在线数据库预测，双荧光素酶报告基因测定证实了 ZFPM2-AS1 与 microR-515（miR-515）或同源盒 A1（HOXA1）之间的相关性。通过定量实时 PCR 和 Western blot 测定确定相关基因的表达。
ZFPM2-AS1 在 RB 组织和细胞系中显著过表达，ZFPM2-AS1 沉默在体外和体内均抑制 RB 细胞的生长和转移。生物信息学网站和双荧光素酶报告基因测定显示，ZFPM2-AS1 可能作为 miR-515 的竞争性内源性 RNA 发挥作用，并与 HOXA1 正相关激活 Wnt/β-catenin 信号通路。
总之，这些数据表明 ZFPM2-AS1 通过介导 miR-515 与 HOXA1 相互作用促进 RB 发展，为 RB 建立了有前途的治疗生物标志物和预后标志物。
