Department of Anesthesiology, Affiliated Hospital of Hebei University, Baoding City, Hebei Province, China.
Histol Histopathol. 2022 Dec;37(12):1213-1226. doi: 10.14670/HH-18-465. Epub 2022 May 6.
Recently, circular RNAs (circRNAs) have been emerging as new regulators in the propofol-induced tumor-suppressive role. Here, we intended to investigate the involvement of circRNA-Mediator of cell motility 1 (circ-MEMO1; hsa_circ_0007385) in propofol role in cancer hallmarks of lung adenocarcinoma (LUAD).
Real-time quantitative PCR and western blotting examined transcriptional and translational levels of circ-MEMO1, microRNA (miR)-485-3p, and NIMA-related kinase-4 (NEK4), and markers of growth and metastasis including E-cadherin, CyclinD1, and Vimentin. Cancer hallmarks were measured by 3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, flow cytometry, 5-ethynyl-2-deoxyuridine assay, and transwell assay. The interaction among circ-MEMO1, miR-485-3p, NEK4 was determined by dual-luciferase reporter assay and Pearson's correlation analysis.
Circ-MEMO1 and NEK4 were high-expressed, and miR-485-3p was low-expressed in LUAD patients and cells; moreover, circ-MEMO1 and NEK4 expression in LUAD cells could be suppressed, whereas miR-485-3p could be elevated with propofol anesthesia. Functionally, propofol restrained cell viability, cell cycle entrance, cell proliferation, migration, and invasion of LUAD cells, accompanied by promoted E-cadherin and depressed CyclinD1 and Vimentin. Coincidently, high circ-MEMO1 was associated with low overall survival of LUAD patients, and overexpressing circ-MEMO1 could overall attenuate propofol effects in LUAD cells. Of note, upregulating miR-485-3p and/or interfering NEK4 could partially countermand the adverse impacts of circ-MEMO1 on propofol's role in LUAD cells. Importantly, circ-MEMO1 acted as a sponge for miR-485-3p to modulate the expression of miR-485-3p-targeted oncogene NEK4.
Promoting the circ-MEMO1-miR-485-3p-NEK4 axis might halt the tumor-inhibiting role of propofol in LUAD cells in vitro, suggesting a potential epigenetic pathway of propofol.
最近，环状 RNA（circRNA）作为一种新的抑瘤因子，在异丙酚诱导的抑瘤作用中逐渐显现出来。在这里，我们旨在研究环状 RNA-细胞运动调节剂 1（circ-MEMO1；hsa_circ_0007385）在肺腺癌（LUAD）中异丙酚作用的癌症特征中的作用。
实时定量 PCR 和 Western blot 检测环状 RNA-MEMO1、微小 RNA（miR）-485-3p 和 NIMA 相关激酶-4（NEK4）的转录和翻译水平，以及生长和转移标志物，包括 E-钙粘蛋白、细胞周期蛋白 D1 和波形蛋白。通过 3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四唑溴盐（MTT）测定法、流式细胞术、5-乙炔基-2-脱氧尿苷（EdU）测定法和 Transwell 测定法测量癌症特征。通过双荧光素酶报告基因测定和 Pearson 相关性分析确定 circ-MEMO1、miR-485-3p 和 NEK4 之间的相互作用。
circ-MEMO1 和 NEK4 在 LUAD 患者和细胞中高表达，miR-485-3p 低表达；此外，异丙酚麻醉可抑制 LUAD 细胞中 circ-MEMO1 和 NEK4 的表达，而 miR-485-3p 的表达可升高。功能上，异丙酚抑制 LUAD 细胞的活力、细胞周期进入、细胞增殖、迁移和侵袭，同时促进 E-钙粘蛋白的表达，抑制细胞周期蛋白 D1 和波形蛋白的表达。同时，circ-MEMO1 与 LUAD 患者的总生存率呈负相关，过表达 circ-MEMO1 可整体减弱异丙酚在 LUAD 细胞中的作用。值得注意的是，上调 miR-485-3p 和/或干扰 NEK4 可部分抵消 circ-MEMO1 对异丙酚在 LUAD 细胞中作用的不利影响。重要的是，circ-MEMO1 作为 miR-485-3p 的海绵，调节 miR-485-3p 靶向癌基因 NEK4 的表达。
促进 circ-MEMO1-miR-485-3p-NEK4 轴可能会阻止异丙酚在体外抑制 LUAD 细胞中的肿瘤作用，提示异丙酚存在潜在的表观遗传途径。
