Department of Hepatobiliary Surgery, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710061, People's Republic of China.
World J Surg Oncol. 2023 Mar 28;21(1):111. doi: 10.1186/s12957-023-02993-z.
Long non-coding RNAs (lncRNAs) perform a vital role during the progression of hepatocellular carcinoma (HCC). Here, we aimed to identify a novel lncRNA involved in HCC development and elucidate the underlying molecular mechanism.
The RT-qPCR and TCGA dataset analysis were applied to explore the expressions of MRVI1-AS1 in HCC tissues and cell lines. Statistical analysis was applied to analyze the clinical significance of MRVI1-AS1 in HCC. The functions of MRVI1-AS1 in HCC cells metastasis and growth were explored by transwell assays, wound healing assay, MTT assay, EdU assay, the intravenous transplantation tumor model, and the subcutaneous xenograft tumor model. Microarray mRNA expression analysis, dual luciferase assays, and actinomycin D treatment were used to explore the downstream target of MRVI1-AS1 in HCC cells. RIP assay was applied to assess the direct interactions between CELF2 and MRVI1-AS1 or SKA1 mRNA. Rescue experiments were employed to validate the functional effects of MRVI1-AS1, CELF2, and SKA1 on HCC cells.
MRVI1-AS1 was found to be dramatically upregulated in HCC and the expression was strongly linked to tumor size, venous infiltration, TNM stage, as well as HCC patients' outcome. Cytological experiments and animal experiments showed that MRVI1-AS1 promoted HCC cells metastasis and growth. Furthermore, SKA1 was identified as the downstream targeted mRNA of MRVI1-AS1 in HCC cells, and MRVI1-AS1 increased SKA1 expression by recruiting CELF2 protein to stabilize SKA1 mRNA. In addition, we found that MRVI1-AS1 expression was stimulated by hypoxia through a HIF-1-dependent manner, which meant that MRVI1-AS was a direct downstream target gene of HIF-1 in HCC.
In a word, our findings elucidated that hypoxia-induced MRVI1-AS1 promotes metastasis and growth of HCC cells via recruiting CELF2 protein to stabilize SKA1 mRNA, pointing to MRVI1-AS1 as a promising clinical application target for HCC therapy.
长非编码 RNA（lncRNA）在肝细胞癌（HCC）的进展中发挥着重要作用。在这里，我们旨在鉴定一种新的参与 HCC 发生发展的 lncRNA，并阐明其潜在的分子机制。
应用 RT-qPCR 和 TCGA 数据集分析来研究 HCC 组织和细胞系中 MRVI1-AS1 的表达。应用统计学分析来分析 MRVI1-AS1 在 HCC 中的临床意义。通过 Transwell 测定、划痕愈合测定、MTT 测定、EdU 测定、静脉移植肿瘤模型和皮下异种移植肿瘤模型来研究 MRVI1-AS1 在 HCC 细胞转移和生长中的作用。微阵列 mRNA 表达分析、双荧光素酶测定和 Actinomycin D 处理用于研究 HCC 细胞中 MRVI1-AS1 的下游靶标。RIP 测定用于评估 CELF2 和 MRVI1-AS1 或 SKA1 mRNA 之间的直接相互作用。进行挽救实验来验证 MRVI1-AS1、CELF2 和 SKA1 对 HCC 细胞的功能影响。
MRVI1-AS1 在 HCC 中显著上调，其表达与肿瘤大小、静脉浸润、TNM 分期以及 HCC 患者的预后强烈相关。细胞实验和动物实验表明，MRVI1-AS1 促进 HCC 细胞转移和生长。此外，在 HCC 细胞中鉴定出 SKA1 是 MRVI1-AS1 的下游靶标 mRNA，MRVI1-AS1 通过募集 CELF2 蛋白稳定 SKA1 mRNA 来增加 SKA1 的表达。此外，我们发现，MRVI1-AS1 的表达通过 HIF-1 依赖性方式受到缺氧的刺激，这意味着 MRVI1-AS1 是 HCC 中 HIF-1 的直接下游靶基因。
总之，我们的研究结果表明，缺氧诱导的 MRVI1-AS1 通过募集 CELF2 蛋白稳定 SKA1 mRNA 促进 HCC 细胞的转移和生长，提示 MRVI1-AS1 作为 HCC 治疗的一个有前途的临床应用靶点。
