敲低长链非编码 RNA ANRIL 通过下调 miR-191 抑制 HepG2 细胞的增殖、迁移和侵袭。
Knockdown long non-coding RNA ANRIL inhibits proliferation, migration and invasion of HepG2 cells by down-regulation of miR-191.
机构信息
Department of Infectious Disease, The Affiliated Hospital of Qingdao University, Qingdao, 266003, Shandong, China.
Department of Gastroenterology, The Affiliated Hospital of Qingdao University, No.16, Jiangsu Road, Qingdao, 266003, Shandong, China.
出版信息
BMC Cancer. 2018 Sep 24;18(1):919. doi: 10.1186/s12885-018-4831-6.
BACKGROUND
Hepatocellular carcinoma (HCC) is a common malignant tumor with high fatality rate. Recent studies reported that up-regulation of long non-coding RNA antisense non-coding RNA in the INK4 locus (lncRNA ANRIL) was found in HCC tissues, and which could affect HCC cells biological processes. However, the potential molecular mechanism of ANRIL in HCC is still unclear. The study aimed to uncover the effect of ANRIL on HepG2 cells growth, migration and invasion.
METHODS
The knockdown expression vectors of ANRIL were transfected into HepG2 cells, and qRT-PCR, CCK-8, flow cytometry, Transwell and western blot assays were performed to analyze the effect of ANRIL on cell proliferation, apoptosis, migration and invasion. The relative expression of miR-191 was then examined in ANRIL knockdown vector transfected cells. These experiments were repeated again for exploring the effect of miR-191 on HepG2 cells. NF-κB and Wnt/β-catenin signaling pathways were examined by using western blot assay.
RESULTS
Knockdown of ANRIL inhibited proliferation, induced apoptosis, meanwhile suppressed migration and invasion of HepG2 cells. Additionally, the results showed that the expression level of miR-191 was down-regulated by ANRIL knockdown in HepG2 cells. Importantly, overexpression of miR-191 reversed the anti-tumor effect of ANRIL on cell proliferation, apoptosis, migration and invasion in HepG2 cells. Besides, we found that ANRIL knockdown inactivated NF-κB and Wnt/β-catenin pathways by regulating miR-191.
CONCLUSIONS
These data demonstrated that ANRIL knockdown suppressed proliferation, migration, invasion, and promoted apoptosis in HepG2 cells by down-regulating miR-191 and inactivating NF-κB and Wnt/β-catenin signaling pathways.
背景
肝细胞癌（HCC）是一种常见的恶性肿瘤，死亡率很高。最近的研究报道，在 HCC 组织中发现长非编码 RNA 反义非编码 RNA 位于 INK4 基因座（lncRNA ANRIL）的上调，它可以影响 HCC 细胞的生物学过程。然而，ANRIL 在 HCC 中的潜在分子机制尚不清楚。本研究旨在揭示 ANRIL 对 HepG2 细胞生长、迁移和侵袭的影响。
方法
将 ANRIL 的敲低表达载体转染到 HepG2 细胞中，然后通过 qRT-PCR、CCK-8、流式细胞术、Transwell 和 Western blot 分析检测 ANRIL 对细胞增殖、凋亡、迁移和侵袭的影响。然后检查 ANRIL 敲低载体转染细胞中 miR-191 的相对表达。再次重复这些实验以探索 miR-191 对 HepG2 细胞的影响。通过 Western blot 分析检测 NF-κB 和 Wnt/β-catenin 信号通路。
结果
下调 ANRIL 抑制 HepG2 细胞的增殖，诱导细胞凋亡，同时抑制细胞迁移和侵袭。此外，结果表明，在 HepG2 细胞中，ANRIL 敲低会下调 miR-191 的表达水平。重要的是，miR-191 的过表达逆转了 ANRIL 对 HepG2 细胞增殖、凋亡、迁移和侵袭的抗肿瘤作用。此外，我们发现 ANRIL 敲低通过调节 miR-191 来抑制 NF-κB 和 Wnt/β-catenin 通路的活性。
结论
这些数据表明，ANRIL 敲低通过下调 miR-191 并抑制 NF-κB 和 Wnt/β-catenin 信号通路来抑制 HepG2 细胞的增殖、迁移、侵袭，并促进细胞凋亡。
Zotero 插件