Department of Hepatology, The First Hospital of Jilin University, Changchun 130021, China.
Department of Hepatobiliary and Pancreas Surgery, The First Hospital of Jilin University, Changchun 130021, China.
Aging (Albany NY). 2020 Jun 18;12(12):11623-11635. doi: 10.18632/aging.103324.
Hepatocellular carcinoma (HCC) is a primary cause of cancer-related deaths globally. While there have been advancements in HCC treatment and diagnosis, incidence and mortality rates continue to rise. One study found that circular RNAs functioned as competing endogenous RNAs, and constructed a gene-based nomogram to estimate overall survival of HCC patients. Previous studies using high-throughput sequencing suggested that hsa_circ_0101145 is abnormally expressed in HCC, but the underlying mechanism is unknown. We performed RT-qPCR to determine hsa_circ_0101145 and miR-548c-3p expression in HCC tissues. We used fluorescence hybridization (FISH) to detect hsa_circ_0101145 expression and hsa_circ_0101145 subcellular localization in HCC tissues. hsa_circ_0101145 expression in HCC cells was selectively regulated. We determined LAMC2 and EMT mRNA and protein levels by RT-qPCR and western blotting analysis, respectively. We employed flow cytometry, and CCK8, Transwell, and wound healing assays to monitor the cell cycle, cell proliferation, invasion, and migration, respectively. We employed dual-luciferase reporter and RNA pulldown assays to verify the relationship among hsa_circ_0101145, miR-548c-3p, and LAMC2. We examined the effects of hsa_circ_0101145 on HCC cell metastasis and proliferation using a subcutaneous xenograft model as well as intravenous tail injection of nude mice. The data demonstrated that hsa_circ_0101145 was significantly upregulated in both HCC tissues and cell lines. High hsa_circ_0101145 expression was correlated with aggressive HCC phenotypes. Downregulation of hsa_circ_0101145 suppressed HCC proliferation as well as metastasis by targeting the miR-548c-3p/LAMC2 axis, which was examined using luciferase reporter and RNA pulldown assays. Silencing of hsa_circ_0101145 suppressed the epithelial-mesenchymal transition in HCC. Downregulation of miR-548c-3p or overexpression of LAMC2 restored migration and proliferation abilities of HCC cells following hsa_circ_0101145 silencing. LAMC2 overexpression reversed miR-548c-3p-induced cell migration and growth inhibition . In summary, the findings illustrated that hsa_circ_0101145 silencing suppressed HCC progression by functioning as an miR-548c-3p sponge to enhance LAMC2 expression. Therefore, hsa_circ_0101145 could be an HCC treatment target.
肝细胞癌 (HCC) 是全球癌症相关死亡的主要原因。尽管 HCC 的治疗和诊断取得了进展，但发病率和死亡率仍在持续上升。一项研究发现环状 RNA 作为竞争性内源性 RNA 发挥作用，并构建了一个基于基因的列线图来估计 HCC 患者的总生存率。先前使用高通量测序的研究表明 hsa_circ_0101145 在 HCC 中异常表达，但潜在机制尚不清楚。我们通过 RT-qPCR 测定 HCC 组织中 hsa_circ_0101145 和 miR-548c-3p 的表达。我们使用荧光杂交 (FISH) 检测 HCC 组织中 hsa_circ_0101145 的表达和 hsa_circ_0101145 的亚细胞定位。HCC 细胞中 hsa_circ_0101145 的表达受到选择性调控。我们通过 RT-qPCR 和 Western blot 分析分别确定 LAMC2 和 EMT mRNA 和蛋白水平。我们采用流式细胞术、CCK8、Transwell 和划痕愈合实验分别监测细胞周期、细胞增殖、侵袭和迁移。我们采用双荧光素酶报告和 RNA 下拉实验验证 hsa_circ_0101145、miR-548c-3p 和 LAMC2 之间的关系。我们通过皮下异种移植模型和裸鼠静脉尾注射研究 hsa_circ_0101145 对 HCC 细胞转移和增殖的影响。数据表明 hsa_circ_0101145 在 HCC 组织和细胞系中均显著上调。高 hsa_circ_0101145 表达与侵袭性 HCC 表型相关。通过靶向 miR-548c-3p/LAMC2 轴，下调 hsa_circ_0101145 可抑制 HCC 的增殖和转移，这通过荧光素酶报告和 RNA 下拉实验得到证实。沉默 hsa_circ_0101145 可抑制 HCC 中的上皮-间充质转化。沉默 hsa_circ_0101145 后，miR-548c-3p 的下调或 LAMC2 的过表达可恢复 HCC 细胞的迁移和增殖能力。LAMC2 的过表达逆转了 miR-548c-3p 诱导的细胞迁移和生长抑制。综上所述，这些发现表明 hsa_circ_0101145 沉默通过作为 miR-548c-3p 的海绵来增强 LAMC2 的表达，从而抑制 HCC 的进展。因此，hsa_circ_0101145 可能成为 HCC 的治疗靶点。
