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长非编码 RNA 长基因间非蛋白编码 RNA 1232 通过调节 miR-370-5p/PIM3 轴促进膀胱癌中细胞的增殖、迁移和侵袭。

Long non-coding RNA long intergenic non-protein coding RNA 1232 promotes cell proliferation, migration and invasion in bladder cancer via modulating miR-370-5p/PIM3 axis.

机构信息

Department of Urology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong, China.

Department of Central Sterile Supply, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, Shandong, China.

出版信息

J Tissue Eng Regen Med. 2022 Jun;16(6):575-585. doi: 10.1002/term.3291. Epub 2022 Mar 26.

Abstract

Increasing evidences have suggested that long non-coding RNAs are critical regulators in the progression of tumor growth. Long intergenic non-protein coding RNA 1232 (LINC01232) was verified as an oncogene in multiple cancers. Nevertheless, its function in bladder cancer (BC) remains to be uncovered. In the current study, we detected LINC01232 expression utilizing quantitative real-time polymerase chain reaction (RT-qPCR) and discovered that LINC01232 was overexpressed in BC cell lines versus normal cell line. Besides, the effect of LINC01232 on BC cell behaviors was measured by colony formation, Cell Counting Kit-8 (CCK-8), transwell, TdT-mediated dUTP Nick-End Labeling and caspase-3/8 activity assays. Functionally, LINC01232 deficiency suppressed cell proliferation, migration and invasion. Next, miR-370-5p was proved to bind with LINC01232 by RNA pull down, RNA-binding protein immunoprecipitation (RIP) and luciferase reporter assays. Furthermore, PIM3 expression was negatively modulated by miR-370-5p and markedly increased in BC cell lines. Moreover, PIM3 silence repressed proliferation, migration and invasion but triggered apoptosis of BC cells. The rescue assays validated that upregulation of PIM3 recovered the effects of LINC01232 silence on the growth of BC cells. To summarize, our study manifested that LINC01232 accelerates BC progression by targeting miR-370-5p/PIM3 axis. Targeting LINC01232 might offer novel insight into BC treatment.

摘要

越来越多的证据表明,长非编码 RNA 是肿瘤生长进展的关键调节因子。长链非编码 RNA 1232(LINC01232)已被证实为多种癌症的癌基因。然而,其在膀胱癌(BC)中的作用仍有待揭示。在本研究中,我们利用实时定量聚合酶链反应(RT-qPCR)检测了 LINC01232 的表达,并发现 LINC01232 在 BC 细胞系中表达上调,而在正常细胞系中表达下调。此外,通过集落形成、Cell Counting Kit-8(CCK-8)、Transwell、TdT 介导的 dUTP 缺口末端标记和 caspase-3/8 活性测定来测量 LINC01232 对 BC 细胞行为的影响。功能上,LINC01232 缺失抑制了细胞增殖、迁移和侵袭。接下来,通过 RNA 下拉、RNA 结合蛋白免疫沉淀(RIP)和荧光素酶报告基因实验证实 miR-370-5p 与 LINC01232 结合。此外,PIM3 表达受 miR-370-5p 负调控,在 BC 细胞系中明显增加。此外,PIM3 沉默抑制了 BC 细胞的增殖、迁移和侵袭,但触发了细胞凋亡。挽救实验验证了上调 PIM3 恢复了 LINC01232 沉默对 BC 细胞生长的影响。总之,我们的研究表明,LINC01232 通过靶向 miR-370-5p/PIM3 轴加速 BC 的进展。靶向 LINC01232 可能为 BC 的治疗提供新的思路。

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