Department of Radiology, Xiangya Hospital, Central South University, Changsha, China.
Department of Infectious Disease, The Third Xiangya Hospital, Central South University, Changsha, China.
Liver Int. 2022 Mar;42(3):682-695. doi: 10.1111/liv.15097. Epub 2021 Nov 29.
BACKGROUND & AIMS: Hepatocellular carcinoma (HCC) is one of the leading causes of tumour-related death. Here, we investigated the molecular mechanism of HCC by studying the function of circ_GLIS2.
Human HCC specimens and cell lines were used. Sanger sequencing, actinomycin D and RNase R treatment were performed to validate circular RNA features of circ_GLIS2. qRT-PCR, western blotting, immunostaining, and IHC were employed to examine levels of circ_GLIS2, GLIS2 mRNA, and EMT-related markers. CCK-8, colony formation, flow cytometry, wound healing assay, and transwell assays were performed to evaluate cancer cell proliferation, apoptosis, migration, and invasion. RIP and RNA pull-down assay were used to validate EIF4A3/GLIS2 mRNA interaction. MSP was performed to measure the methylation status of GLIS2 promoter. Nude mouse xenograft model was used to examine tumour growth and metastasis in vivo.
Circ_GLIS2 and linear GLIS2 mRNA were reduced in human HCC tissues and cells. Their low levels correlated with a poor survival rate of HCC patients. Overexpression of circ_GLIS2 and GLIS2 suppressed HCC cell proliferation, migration, and invasion but promoted cell apoptosis. GLIS2 promoter region was hypermethylated in HCC cells. EIF4A3 was directly bound with GLIS2 mRNA and promoted circ_GLIS2/GLIS2 expression. Moreover, overexpression of circ_GLIS2 restrained HCC tumour growth and metastasis in vivo.
Circ_GLIS2 suppresses HCC growth and metastasis by inhibiting cell proliferation, migration, and invasion, but promoting cell apoptosis. These findings provide molecular insights into the mechanism of HCC and indicate that circ_GLIS2 could serve as a diagnosis marker or therapeutic target for HCC.
肝细胞癌（HCC）是肿瘤相关死亡的主要原因之一。在此，我们通过研究 circ_GLIS2 的功能来研究 HCC 的分子机制。
使用人 HCC 标本和细胞系。进行 Sanger 测序、放线菌素 D 和 RNase R 处理以验证 circ_GLIS2 的环状 RNA 特征。使用 qRT-PCR、western blot、免疫染色和 IHC 检测 circ_GLIS2、GLIS2 mRNA 和 EMT 相关标记物的水平。进行 CCK-8、集落形成、流式细胞术、划痕愈合实验和 Transwell 实验以评估癌细胞增殖、凋亡、迁移和侵袭。使用 RIP 和 RNA 下拉实验验证 EIF4A3/GLIS2 mRNA 相互作用。MSP 用于测量 GLIS2 启动子的甲基化状态。裸鼠异种移植模型用于体内检测肿瘤生长和转移。
circ_GLIS2 和线性 GLIS2 mRNA 在人 HCC 组织和细胞中减少。它们的低水平与 HCC 患者的生存率差相关。circ_GLIS2 和 GLIS2 的过表达抑制 HCC 细胞增殖、迁移和侵袭，但促进细胞凋亡。HCC 细胞中 GLIS2 启动子区域发生高甲基化。EIF4A3 直接与 GLIS2 mRNA 结合并促进 circ_GLIS2/GLIS2 表达。此外，circ_GLIS2 的过表达抑制体内 HCC 肿瘤生长和转移。
circ_GLIS2 通过抑制细胞增殖、迁移和侵袭，促进细胞凋亡来抑制 HCC 的生长和转移。这些发现为 HCC 的机制提供了分子见解，并表明 circ_GLIS2 可以作为 HCC 的诊断标志物或治疗靶标。
