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Abstract：
Long non‑coding RNAs (lncRNAs) are common in the human body. Misregulated lncRNA expression can cause a variety of diseases in the human body. The present study aimed to investigate the effect of lncRNA differentiation antagonizing non‑protein‑coding RNA (DANCR) on glioma proliferation and autophagy through the microRNA (miR)‑33b/distal‑less homeobox 6 (DLX6)/autophagy‑related 7 (ATG7) axis. Reverse transcription‑quantitative PCR was used to detect DANCR and miR‑33b expression. Cell Counting Kit‑8 assay and flow cytometry were used to detect cell proliferation and apoptosis, respectively. Transmission electron microscopy was used to determine the autophagy level by observing intracellular autophagosomes. A western blot assay was used to detect protein expression levels and determine the level of autophagy in different cells. The binding sites of miR‑33b and DANCR or DLX6 were detected using a dual‑luciferase reporter assay. A chromatin immunoprecipitation assay confirmed DLX6 as a transcript of ATG7. In vivo tumorigenesis of glioma cells was validated in nude mice. DANCR and DLX6 were highly expressed in glioma cells, while miR‑33b showed low expression in glioma cells. DANCR reduced the targeted binding of miR‑33b to DLX6 by sponging miR‑33b. The result verified that DANCR could promote ATG7 protein expression through miR‑33b/DLX6, promote intracellular autophagy and proliferation and reduce apoptosis. The present study identified the role of the DANCR/miR‑33b/DLX6/ATG7 axis in regulating autophagy, proliferation, and apoptosis in glioma cells, providing new ideas for glioma treatment.
摘要：
长链非编码RNA（lncRNA）在人体中很常见。lncRNA表达失调可引起人体多种疾病。本研究旨在研究lncRNA分化拮抗非蛋白编码RNA（DANCR）通过microRNA（miR）-33b/非远端同源盒6（DLX6）/自噬相关7（ATG7）轴对神经胶质瘤增殖和自噬的影响。逆转录-定量PCR用于检测DANCR和miR-33b的表达。细胞计数试剂盒-8测定和流式细胞术用于检测细胞增殖和凋亡，分别。采用透射电镜观察细胞内自噬体的自噬水平。采用westernblot法检测不同细胞的蛋白表达水平并测定自噬水平。miR-33b和DANCR或DLX6的结合位点使用双荧光素酶报告基因测定进行检测。染色质免疫沉淀测定证实DLX6为ATG7的转录物。在裸鼠中验证了神经胶质瘤细胞的体内肿瘤发生。DANCR和DLX6在胶质瘤细胞中高表达，而miR-33b在神经胶质瘤细胞中显示低表达。DANCR通过使用miR‑33b减少miR‑33b与DLX6的靶向结合。结果证实DANCR可通过miR‑33b/DLX6促进ATG7蛋白表达，促进细胞内自噬和增殖，减少细胞凋亡。本研究确定了DANCR/miR-33b/DLX6/ATG7轴在调节自噬中的作用，扩散,和神经胶质瘤细胞的凋亡，为脑胶质瘤的治疗提供新思路。
