Suzhou Medical College of Soochow University, Suzhou, China.
Department of General Surgery (Hepatobiliary and Pancreatic Surgery), The First Affiliated Hospital of Gannan Medical University, Ganzhou, China.
Neoplasma. 2023 Jun;70(3):361-374. doi: 10.4149/neo_2023_230323N162.
Hepatocellular carcinoma (HCC) is a malignant tumor, which seriously threatens the life of patients. LncRNA SLC7A11-AS1 was reported to be abnormally expressed in HCC. Here, the functions and relative molecular regulatory mechanism of SLC7A11-AS1 in HCC were investigated. Nude mice and HCC cells were used as the experimental subjects. Knockdown or overexpression of exogenous genes was conducted in HCC cells. RT-qPCR, IHC, and western blot were employed to evaluate the abundance of genes and proteins. The malignant behaviors were evaluated using CCK-8, clone formation, wound-healing, and Transwell. The locations of SLC7A11-AS1 and KLF9 in cells were determined by FISH and IF assays. The total m6A level was evaluated by dot-blot assay. m6A modification of SLC7A11-AS1 was detected using RNA MeRIP. The interactions among molecules were validated by RIP, ChIP, dual luciferase reporter assay, and co-IP. SLC7A11-AS1 was elevated apparently in HCC cells and HCC tissues from mice. SLC7A11-AS1 silencing could suppress HCC progression, which was validated in in vivo and in vitro experiments. Furthermore, METTL3 mediated m6A modification of SLC7A11-AS1 to elevate its expression. In addition, SLC7A11-AS1 downregulated KLF9 expression by affecting STUB1-mediated ubiquitination degradation and KLF9 enhanced PHLPP2 expression to inactivate the AKT pathway. Eventually, rescue experiments revealed that KLF9 knockdown abolished SLC7A11-AS1 silencing-mediated suppression of HCC progression in vivo and in vitro. Our results unveiled that m6A-modified SLC7A11-AS1 promoted HCC progression by regulating the STUB1/KLF9/PHLPP2/AKT axis, indicating that targeting SLC7A11-AS1 might alleviate HCC progression.
肝细胞癌（HCC）是一种恶性肿瘤，严重威胁着患者的生命。有研究报道长链非编码 RNA SLC7A11-AS1 在 HCC 中异常表达。本研究旨在探讨 SLC7A11-AS1 在 HCC 中的功能及其相关分子调控机制。裸鼠和 HCC 细胞被用作实验对象。在 HCC 细胞中外源基因进行敲低或过表达。采用 RT-qPCR、免疫组化和 Western blot 检测基因和蛋白的丰度。使用 CCK-8、克隆形成、划痕愈合和 Transwell 实验评估恶性行为。通过 FISH 和 IF 实验确定 SLC7A11-AS1 和 KLF9 在细胞中的位置。采用 dot-blot assay 评估总 m6A 水平。通过 RNA MeRIP 检测 SLC7A11-AS1 的 m6A 修饰。通过 RIP、ChIP、双荧光素酶报告基因检测和 co-IP 验证分子间的相互作用。结果显示，SLC7A11-AS1 在 HCC 细胞和小鼠 HCC 组织中明显上调。SLC7A11-AS1 沉默可抑制 HCC 的进展，在体内和体外实验中得到验证。此外，METTL3 通过 m6A 修饰 SLC7A11-AS1 来提高其表达。此外，SLC7A11-AS1 通过影响 STUB1 介导的泛素化降解来下调 KLF9 表达，KLF9 增强 PHLPP2 表达以激活 AKT 通路。最终，挽救实验表明，体内和体外 KLF9 敲低消除了 SLC7A11-AS1 沉默对 HCC 进展的抑制作用。综上所述，m6A 修饰的 SLC7A11-AS1 通过调节 STUB1/KLF9/PHLPP2/AKT 轴促进 HCC 进展，表明靶向 SLC7A11-AS1 可能减轻 HCC 进展。
