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长链非编码 RNA SLCO4A1-AS1 通过与 miR-150-3p 结合并正向调控 SLCO4A1 来调节结肠癌细胞干性。

LncRNA SLCO4A1-AS1 modulates colon cancer stem cell properties by binding to miR-150-3p and positively regulating SLCO4A1.

机构信息

Department of Gastrointestinal Surgery, The Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University, Huai'an, 223300, PR China.

Department of Hematology, The Affiliated Huai'an No.1 People's Hospital of Nanjing Medical University, Huai'an, 223300, PR China.

出版信息

Lab Invest. 2021 Jul;101(7):908-920. doi: 10.1038/s41374-021-00577-7. Epub 2021 May 6.

DOI:10.1038/s41374-021-00577-7
PMID:33958701
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8214995/
Abstract

Long non-coding RNAs (lncRNAs) play important roles in a range of different human cancers. However, the role of lncRNA solute carrier organic anion transporter family member 4A1-AS1 (SLCO4A1-AS1) in colon cancer remains enigmatic. Hence, we aimed to explore the specific role of SLCO4A1-AS1 in colon cancer stem cells. Colon cancer-related differentially expressed lncRNA and mRNA were screened using microarray-based analysis, and the expression of SLCO4A1-AS1 and SLCO4A1 in colon cancer tissues was determined using reverse transcription quantitative polymerase chain reaction and western blot analysis. The interaction among SLCO4A1-AS1, microRNA-150-3p (miR-150-3p) and SLCO4A1 was verified using dual-luciferase reporter assay, RNA immunoprecipitation and RNA pull-down. Moreover, SLCO4A1-AS1, miR-150-3p and/or SLCO4A1 were overexpressed or depleted in colon cancer cells to detect their effects on migration, invasion, sphere formation, apoptosis and tumorigenesis abilities of colon cancer stem CD133CD44 cells using both in vitro and in vivo assays. SLCO4A1-AS1 and SLCO4A1 were screened as the differentially expressed lncRNA and mRNA in colon cancer tissues. SLCO4A1-AS1 was confirmed to competitively bind to miR-150-3p to elevate SLCO4A1 expression. Moreover, knockdown of SLCO4A1-AS1 decreased SLCO4A1 expression, thus inhibiting cell migration, invasion, sphere formation, and tumorigenesis abilities and enhancing the apoptosis of CD133CD44 cells. Collectively, these findings provide evidence demonstrating that depleting SLCO4A1-AS1 competitively binds to miR-150-3p, which downregulates SLCO4A1 expression, thus hindering colon cancer progression.

摘要

长链非编码 RNA（lncRNA）在多种人类癌症中发挥重要作用。然而，溶质载体有机阴离子转运家族成员 4A1-AS1（SLCO4A1-AS1）在结肠癌中的作用仍然是个谜。因此，我们旨在探讨 SLCO4A1-AS1 在结肠癌干细胞中的特定作用。通过基于微阵列的分析筛选结肠癌相关差异表达的 lncRNA 和 mRNA，并通过逆转录定量聚合酶链反应和 Western blot 分析确定结肠癌组织中 SLCO4A1-AS1 和 SLCO4A1 的表达。通过双荧光素酶报告基因检测、RNA 免疫沉淀和 RNA 下拉实验验证 SLCO4A1-AS1、microRNA-150-3p（miR-150-3p）和 SLCO4A1 之间的相互作用。此外，通过体外和体内实验检测 SLCO4A1-AS1、miR-150-3p 和/或 SLCO4A1 在结肠癌 CD133CD44 细胞中的过表达或耗竭对其迁移、侵袭、球体形成、凋亡和肿瘤发生能力的影响。SLCO4A1-AS1 和 SLCO4A1 被筛选为结肠癌组织中差异表达的 lncRNA 和 mRNA。证实 SLCO4A1-AS1 可与 miR-150-3p 竞争性结合，从而提高 SLCO4A1 的表达。此外，SLCO4A1-AS1 的敲低降低了 SLCO4A1 的表达，从而抑制 CD133CD44 细胞的迁移、侵袭、球体形成和肿瘤发生能力，并增强其凋亡。综上所述，这些发现提供了证据表明，耗竭 SLCO4A1-AS1 竞争性结合 miR-150-3p，下调 SLCO4A1 的表达，从而阻碍结肠癌的进展。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/33d15c1fe22a/41374_2021_577_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/1e0b4797e909/41374_2021_577_Fig1_HTML.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/0bed5c46a23c/41374_2021_577_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/f8d2ecbeb3e4/41374_2021_577_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/526dd0be73f8/41374_2021_577_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/3e3becf660e8/41374_2021_577_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/33d15c1fe22a/41374_2021_577_Fig8_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/1e0b4797e909/41374_2021_577_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/e968583222d2/41374_2021_577_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/6d790e73f139/41374_2021_577_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/0bed5c46a23c/41374_2021_577_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/f8d2ecbeb3e4/41374_2021_577_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/526dd0be73f8/41374_2021_577_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/3e3becf660e8/41374_2021_577_Fig7_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ac8d/8214995/33d15c1fe22a/41374_2021_577_Fig8_HTML.jpg

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