Department of Hepatobiliary Sugery, Shengli Oilfield Central Hospital, Dongying, China.
Department of Sales, Shanghai Topgen Biopharm Company Ltd, shanghai, china.
Bioengineered. 2022 Jan;13(1):1346-1358. doi: 10.1080/21655979.2021.2017565.
Long noncoding RNA muskelin 1 antisense RNA (MKLN1-AS) acted as an oncogenic regulator in hepatocellular carcinoma (HCC). This study was performed to investigate the functional mechanism of MKLN1-AS. MKLN1-AS, microRNA-22-3p (miR-22-3p) and ETS Proto-Oncogene 1 (ETS1) levels were examined using reverse transcription-quantitative polymerase-chain reaction. Protein expression was detected by Western blot. The target relation was analyzed by dual-luciferase reporter assay, RNA immunoprecipitation assay and RNA pull-down assay. Cell proliferation ability was determined through cell counting kit-8 assay, colony formation assay and ethylenediurea assay. Angiogenesis was examined by tube formation assay. Cell migration and invasion were assessed via transwell assay. In vivo research was conducted by xenograft tumor model in nude mice. MKLN1-AS was upregulated in HCC tissues and cells. ETS1 promoted the ETS1 expression by binding to the 582-596 sites. Silence of MKLN1-AS suppressed cell growth, angiogenesis, migration, and invasion. MKLN1-AS interacted with miR-22-3p in HCC cells. The function of MKLN1-AS downregulation was relieved by miR-22-3p inhibition in HCC cells. ETS1 was validated as a target of miR-22-3p, and MKLN1-AS upregulated the ETS1 expression by sponging miR-22-3p. Overexpression of miR-22-3p retarded HCC progression by downregulating the level of ETS1. Tumor growth in vivo was also enhanced by MKLN1-AS through the regulation of miR-22-3p/ETS1 axis. These data demonstrated that ETS1-mediated MKLN1-AS contributed to the malignant phenotypes of HCC cells via depending on the miR-22-3p/ETS1 regulatory axis.
长链非编码 RNA 肌球蛋白 1 反义 RNA（MKLN1-AS）在肝细胞癌（HCC）中作为致癌调节剂发挥作用。本研究旨在探讨 MKLN1-AS 的功能机制。采用逆转录定量聚合酶链反应检测 MKLN1-AS、microRNA-22-3p（miR-22-3p）和 ETS 原癌基因 1（ETS1）水平。通过 Western blot 检测蛋白表达。通过双荧光素酶报告基因检测、RNA 免疫沉淀试验和 RNA 下拉试验分析靶关系。通过细胞计数试剂盒-8 试验、集落形成试验和乙二醛试验测定细胞增殖能力。通过管形成试验检测血管生成。通过 Transwell 试验评估细胞迁移和侵袭。通过裸鼠异种移植肿瘤模型进行体内研究。MKLN1-AS 在 HCC 组织和细胞中上调。ETS1 通过结合 582-596 位点促进 ETS1 的表达。沉默 MKLN1-AS 抑制 HCC 细胞的生长、血管生成、迁移和侵袭。MKLN1-AS 在 HCC 细胞中与 miR-22-3p 相互作用。MKLN1-AS 下调在 HCC 细胞中通过抑制 miR-22-3p 缓解了 miR-22-3p 抑制的功能。ETS1 被验证为 miR-22-3p 的靶基因，MKLN1-AS 通过海绵 miR-22-3p 上调 ETS1 的表达。过表达 miR-22-3p 通过下调 ETS1 水平抑制 HCC 进展。体内肿瘤生长也通过 miR-22-3p/ETS1 轴的调节被 MKLN1-AS 增强。这些数据表明，ETS1 介导的 MKLN1-AS 通过依赖于 miR-22-3p/ETS1 调节轴促进 HCC 细胞的恶性表型。
