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LINC00839/miR-144-3p/WTAP(WT1 相关蛋白)轴参与调控肝细胞癌进展。

LINC00839/miR-144-3p/WTAP (WT1 Associated protein) axis is involved in regulating hepatocellular carcinoma progression.

机构信息

Department of Radiotherapy, Affiliated Hospital of Nantong University, Nantong, China.

Medical College of Southeast University, Nanjing, China.

出版信息

Bioengineered. 2021 Dec;12(2):10849-10861. doi: 10.1080/21655979.2021.1990578.

Abstract

The present work aimed to explore LINC00839 expression level and its function in hepatocellular carcinoma (HCC), and identify the downstream molecular mechanisms. qRT-PCR (Real-Time Quantitative Reverse Transcription PCR) and western blot were employed to detect mRNA and protein levels. Functional investigations were performed by flow cytometric-based apoptosis assay, CCK8 (Cell Counting Kit-8) assay, clone formation assay, Transwell migration and invasion assay. Functional interactions between LINC00839 and miR-144-3p or miR-144-3p and WTAP were validated by dual luciferase reporter assay. siRNA (small interfering RNA) was used for LINC00839 silencing, and microRNA mimic or inhibitor were employed to modulate miR-144-3p activity. LINC00839 was upregulated in HCC cells and tissues. Silencing LINC00839 suppressed the proliferation, invasion, migration of HCC cells and induced apoptosis. Additionally, LINC00839 served as a sponge to negatively impact on miR-144-3p activity, which contributed to the high expression of WTAP (WT1 Associated Protein) and the malignant phenotype of HCC cells. Our study revealed an oncogenic role of LINC00839 in HCC, and identified miR-144-3p/WTAP axis as downstream effectors mediating the oncogenic function of LINC00839. LINC00839 might serve as a potential therapeutic target and prognostic marker for HCC.

摘要

本研究旨在探索 LINC00839 在肝细胞癌(HCC)中的表达水平及其功能,并鉴定其下游分子机制。采用 qRT-PCR(实时定量逆转录 PCR)和 Western blot 检测 mRNA 和蛋白水平。通过流式细胞术检测细胞凋亡、CCK8 法检测细胞增殖、克隆形成实验、Transwell 迁移和侵袭实验进行功能研究。通过双荧光素酶报告实验验证 LINC00839 与 miR-144-3p 或 miR-144-3p 与 WTAP 之间的功能相互作用。采用 siRNA(小干扰 RNA)沉默 LINC00839,利用 microRNA 模拟物或抑制剂调节 miR-144-3p 的活性。LINC00839 在 HCC 细胞和组织中呈上调表达。沉默 LINC00839 抑制 HCC 细胞的增殖、侵袭和迁移,并诱导细胞凋亡。此外,LINC00839 作为 miR-144-3p 的海绵,负调控 miR-144-3p 的活性,导致 WTAP(WT1 相关蛋白)表达上调,进而促进 HCC 细胞的恶性表型。本研究揭示了 LINC00839 在 HCC 中的致癌作用,并确定了 miR-144-3p/WTAP 轴作为介导 LINC00839 致癌功能的下游效应子。LINC00839 可能成为 HCC 的潜在治疗靶点和预后标志物。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5afd/8809969/9fc174c42819/KBIE_A_1990578_F0001_OC.jpg

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