LINC02154 通过增强 SPC24 启动子活性和激活 PI3K-AKT 信号通路促进肝癌的增殖和转移。

LINC02154 promotes the proliferation and metastasis of hepatocellular carcinoma by enhancing SPC24 promoter activity and activating the PI3K-AKT signaling pathway.

机构信息

Key Laboratory of Molecular Biology for Infectious Diseases (Ministry of Education), Institute for Viral Hepatitis, Department of Infectious Diseases, The Second Affiliated Hospital, Chongqing Medical University, Yi Xue Yuan Road, Chongqing, 400016, China.

Department of Laboratory Medicine, The First People's Hospital of Zunyi City (The Third Affiliated Hospital of Zunyi Medical University), Zunyi, 563000, Guizhou, China.

出版信息

Cell Oncol (Dordr). 2022 Jun;45(3):447-462. doi: 10.1007/s13402-022-00676-7. Epub 2022 May 11.

DOI:10.1007/s13402-022-00676-7

PMID:35543858

Abstract

BACKGROUND

Abnormal expression of long non-coding RNAs (lncRNAs) has been associated with the initiation and progression of hepatocellular carcinoma but, as yet, the clinicopathologic significance and potential role of Linc02154 in HCC remains to be determined. Here, we aimed to investigate the potential role and mode of action of Linc02154 in HCC.

METHODS

The expression of Linc02154 in 20 pairs of HCC/normal tissues and 7 HCC cell lines was detected by qRT-PCR. The localization of Linc02154 in HCC cells was detected using fluorescence in situ hybridization and nuclear-plasma separation assays. MTS, EdU incorporation, colony formation, flow cytometry, scratch wound-healing and transwell assays were performed to assess the role of Linc02154 in HCC cell proliferation, migration and invasion in vitro, and BALB/c nude mice xenografts were used to evaluate its role in vivo. RNA sequencing and Western blotting were used to evaluate the regulatory effect of Linc02154 on SPC24 gene expression. A dual-luciferase reporter assay was used to assess a putative interaction of Linc02154 with the SPC24 promoter.

RESULTS

We identified a new lncRNA, Linc02154, that is highly expressed in HCC cells and tissues of patients with a poor overall survival. Functional experiments revealed that exogenous Linc02154 expression in MHCC-97H and SK-Hep1 cells promoted their proliferation, migration and invasion in vitro and their tumorigenesis in vivo. Using a dual luciferase reporter assay we found that Linc02154 can enhance SPC24 promoter (-500 bp ~ -1000 region) activity. Exogenous over-expression of Linc02154 led to up-regulation of SPC24 by activating PI3K/AKT and its downstream signals, including cell cycle progression and EMT-associated gene expression.

CONCLUSION

Our data suggest that Linc02154 may serve as a valuable biomarker of HCC and as a potential therapeutic target.

摘要

背景

长链非编码 RNA（lncRNAs）的异常表达与肝细胞癌的发生和发展有关，但 Linc02154 在 HCC 中的临床病理意义和潜在作用尚未确定。在这里，我们旨在研究 Linc02154 在 HCC 中的潜在作用和作用模式。

方法

通过 qRT-PCR 检测 20 对 HCC/正常组织和 7 种 HCC 细胞系中 Linc02154 的表达。通过荧光原位杂交和核质分离试验检测 Linc02154 在 HCC 细胞中的定位。MTS、EdU 掺入、集落形成、流式细胞术、划痕愈合和 Transwell 测定用于评估 Linc02154 在 HCC 细胞体外增殖、迁移和侵袭中的作用，BALB/c 裸鼠异种移植用于评估其体内作用。RNA 测序和 Western blot 用于评估 Linc02154 对 SPC24 基因表达的调节作用。双荧光素酶报告基因检测用于评估 Linc02154 与 SPC24 启动子的潜在相互作用。

结果

我们鉴定了一种新的 lncRNA，Linc02154，其在 HCC 细胞和患者的组织中表达水平较高，且总体生存较差。功能实验表明，外源性 Linc02154 在 MHCC-97H 和 SK-Hep1 细胞中的表达促进了它们在体外的增殖、迁移和侵袭以及体内的肿瘤发生。通过双荧光素酶报告基因检测，我们发现 Linc02154 可以增强 SPC24 启动子（-500 bp~-1000 区）的活性。外源性过表达 Linc02154 通过激活 PI3K/AKT 及其下游信号，包括细胞周期进程和 EMT 相关基因表达，导致 SPC24 的上调。