Department of Neurosurgery, The First Affiliated Hospital of Nanchang University, Nanchang, China.
Eur Rev Med Pharmacol Sci. 2019 Jan;23(1):241-252. doi: 10.26355/eurrev_201901_16770.
Long noncoding RNAs (lncRNAs) serve as important regulators of diverse types of cancer, including glioma. Nevertheless, their precise roles in cancers remain sufficiently unexplored.
Quantitative Reverse Transcription-Polymerase Chain Reaction (qRT-PCR) was used to determine the levels of HOMEOBOX A11 antisense RNA (HOXA11-AS) and miR-130a-5p in glioma tissues and cell lines. Short hairpin RNAs (shRNAs) targeting HOXA11-AS or pcDNA3.1 were transfected into cells via a vector encoding HOXA11-AS to decrease or increase the level of HOXA11-AS. Cell Counting Kit-8 (CCK-8), colony formation, wound healing, flow cytometry and transwell assays were applied to assess the role of HOXA11-AS in glioblastoma cell growth, apoptosis and aggressiveness. The expression of N-cadherin and E-cadherin was determined using immunofluorescence staining. The expression of high-mobility group protein B2 (HMGB2) was determined using Western blot analysis in vitro and immunohistochemistry (IHC) staining in vivo. The direct target of HOXA11-AS and miR-130a-5p was confirmed using the Luciferase reporter assay. Glioblastoma cells were subcutaneously implanted into nude mice to determine the role of HOXA11-AS in tumor growth in vivo.
In the current study, we demonstrated that the lncRNA HOXA11-AS was overexpressed in glioma. The overexpression of HOXA11-AS was correlated with advanced stages of glioma and poor prognosis. Downregulating HOXA11-AS expression significantly suppressed the proliferation, migration and invasion of glioma cells and increased their apoptosis. The growth of glioma cells in vitro was also suppressed by the downregulation of HOXA11-AS. Finally, we revealed that HOXA11-AS exerted its oncogenic effects by binding to miR-130a-5p, thereby neutralizing the suppressive effect of miR-130a-5p on HMGB2.
Our results demonstrate that HOXA11-AS regulates the growth and metastasis of glioma by targeting the miR-130a-5p-HMGB2 signaling axis.
长链非编码 RNA（lncRNA）作为多种类型癌症的重要调节因子，包括神经胶质瘤。然而，它们在癌症中的确切作用仍未得到充分探索。
采用实时定量聚合酶链反应（qRT-PCR）检测神经胶质瘤组织和细胞系中同源盒 A11 反义 RNA（HOXA11-AS）和 miR-130a-5p 的水平。通过载体转染靶向 HOXA11-AS 的短发夹 RNA（shRNA）或 pcDNA3.1 来降低或增加 HOXA11-AS 的水平。细胞计数试剂盒-8（CCK-8）、集落形成、划痕愈合、流式细胞术和 Transwell 分析用于评估 HOXA11-AS 在神经胶质瘤细胞生长、凋亡和侵袭中的作用。用免疫荧光染色法测定 N-钙粘蛋白和 E-钙粘蛋白的表达。用 Western blot 分析体外和免疫组织化学（IHC）染色体内测定高迁移率族蛋白 B2（HMGB2）的表达。用荧光素酶报告基因检测法证实 HOXA11-AS 和 miR-130a-5p 的直接靶标。将神经胶质瘤细胞皮下植入裸鼠体内，以确定 HOXA11-AS 在体内肿瘤生长中的作用。
在本研究中，我们证明了长链非编码 RNA HOXA11-AS 在神经胶质瘤中过度表达。HOXA11-AS 的过表达与神经胶质瘤的晚期和不良预后相关。下调 HOXA11-AS 表达显著抑制神经胶质瘤细胞的增殖、迁移和侵袭，并增加其凋亡。HOXA11-AS 的下调也抑制了神经胶质瘤细胞的体外生长。最后，我们揭示了 HOXA11-AS 通过与 miR-130a-5p 结合发挥致癌作用，从而中和 miR-130a-5p 对 HMGB2 的抑制作用。
我们的研究结果表明，HOXA11-AS 通过靶向 miR-130a-5p-HMGB2 信号通路调节神经胶质瘤的生长和转移。
