Department of Respiratory and Critical Care Medicine, Puren Hospital Affiliated to Wuhan University of Science and Technology, Wuhan, China.
Department of Respiratory and Critical Care Medicine, Renmin Hospital of Wuhan University, Wuhan, China.
Bioengineered. 2022 Apr;13(4):10679-10690. doi: 10.1080/21655979.2022.2065946.
Lung adenocarcinoma (LUAD) is a subtype of lung cancer, and therapy remains a great challenge. A growing body of evidence shows that long-chain non-coding RNAs (lncRNAs) play an important role in the occurrence and development of LUAD. This study investigated the roles and mechanisms of action of EBLN3P in LUAD. The bioinformatics software starBase and TargetScan were used to predict the binding sites of the lncRNA endogenous born avirus-like nucleoprotein (EBLN3P) and microRNA (miR)-655-3p in LUAD. The regulatory role of EBLN3P and miR-655-3p in cell proliferation was verified through the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2 H-tetrazolium bromide (MTT) assay. The binding sites between EBLN3P, miR-655-3p, and B-cell lymphoma-2 (Bcl-2) were assessed using dual-luciferase reporter assay, western blotting, and quantitative reverse transcription polymerase chain reaction (qRT-PCR). Flow cytometry (FCM) was performed to analyze the apoptotic rates of A549 cells after transfection. The results revealed that EBLN3P was upregulated, whereas miR-655-3p was downregulated in LUAD cell lines (A549 and NCI-H23). Bioinformatics analysis and dual-luciferase reporter assays indicated that EBLN3P interacted with miR-655-3p. Knockdown of EBLN3P notably inhibited the bioactivity and induced apoptosis in A549 cells by upregulating miR-655-3p. Mechanistically, miR-655-3p inhibits cell viability and induces apoptosis by inhibiting Bcl-2 expression. The high expression of Bcl-2 reversed the impact of miR-655-3p on the inhibition of cell bioactivity and induction of apoptosis in A549 cells. In conclusion, this study demonstrated that EBLN3P silencing inhibits bioactivity and induces apoptosis via the miR-655-3p/Bcl-2 axis, providing a potential therapeutic target for lung adenocarcinoma.
肺腺癌（LUAD）是肺癌的一种亚型，治疗仍然是一个巨大的挑战。越来越多的证据表明，长链非编码 RNA（lncRNA）在 LUAD 的发生和发展中起着重要作用。本研究探讨了 EBLN3P 在 LUAD 中的作用和作用机制。生物信息学软件 starBase 和 TargetScan 用于预测 lncRNA 内源性 born 类病毒核蛋白（EBLN3P）和 microRNA（miR）-655-3p 在 LUAD 中的结合位点。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐（MTT）测定法验证 EBLN3P 和 miR-655-3p 对细胞增殖的调节作用。通过双荧光素酶报告基因检测、Western blot 和实时定量逆转录聚合酶链反应（qRT-PCR）评估 EBLN3P、miR-655-3p 和 B 细胞淋巴瘤-2（Bcl-2）之间的结合位点。流式细胞术（FCM）用于分析转染后 A549 细胞的凋亡率。结果表明，EBLN3P 在 LUAD 细胞系（A549 和 NCI-H23）中上调，而 miR-655-3p 下调。生物信息学分析和双荧光素酶报告基因检测表明，EBLN3P 与 miR-655-3p 相互作用。EBLN3P 敲低通过上调 miR-655-3p 显著抑制 A549 细胞的生物活性并诱导细胞凋亡。机制上，miR-655-3p 通过抑制 Bcl-2 表达抑制细胞活力并诱导细胞凋亡。Bcl-2 的高表达逆转了 miR-655-3p 对 A549 细胞生物活性抑制和诱导细胞凋亡的影响。总之，本研究表明，EBLN3P 沉默通过 miR-655-3p/Bcl-2 轴抑制生物活性并诱导细胞凋亡，为肺腺癌提供了一个潜在的治疗靶点。
