长链非编码 RNA KCNQ1OT1 通过 miR-145-5p/ARF6 轴促进胃癌的进展。
Long non-coding RNA KCNQ1OT1 promotes the progression of gastric cancer via the miR-145-5p/ARF6 axis.
机构信息
Department of Cardiothoracic Surgery, Zhuhai People's Hospital (Zhuhai hospital affiliated with Jinan University), Zhuhai City, Guangdong Province, China.
出版信息
J Gene Med. 2021 May;23(5):e3330. doi: 10.1002/jgm.3330. Epub 2021 Apr 6.
BACKGROUND
Long non-coding RNA KCNQ1 opposite strand/antisense transcript one gene (KCNQ1OT1) has been reported to be involved in the progression of many types of human cancer, whereas its role in gastric cancer (GC) remains unknown. The present study aimed to investigate the role of KCNQ1OT1 in GC.
METHODS
In total, 25 GC tissues and adjacent normal tissues were collected. The expression of KCNQ1OT1, miR-145-5p and ARF6 in GC tissues and cell lines was detected by quantitative reverse transcriptase-polymerase chain reaction or western blotting. Bioinformatics analysis and a dual luciferase reporter assay were performed to determine the relationship between KCNQ1OT1 and miR-145-5p or miR-145-5p and ARF6. Gain- and loss-of function of KCNQ1OT1 and miR-145-5p were achieved to confirm their roles in GC cells. Cell counting kit-8, colony formation and flow cytometry assays were used to evaluate cell viability, proliferation and apoptosis. A xenograft tumor model was established with BGC803 tumor cells transfected with sh-KCNQ1OT1 or empty vector to determine the role of LINC01089 in vivo.
RESULTS
The expression levels of KCNQ1OT1 were markedly elevated in GC tissues and cells. Knockdown of KCNQ1OT1 inhibited GC tumor growth, reduced GC cell viability and colony formation, and induced GC cell apoptosis. The expression levels of miR-145-5p were significantly decreased in GC cells and correlated with the expression of KCNQ1OT1 in GC tumors. Moreover, KCNQ1OT1 directly binds with miR-145-5p, which is targeting ARF6. Knockdown of KCNQ1OT1 increased the expression levels of miR-145-5p. Inhibition of miR-145-5p increased the expression levels of KCNQ1OT1 and also attenuated the effects of knockdown of KCNQ1OT1 on the viability, proliferation and apoptosis of GC cells. In addition, overexpression of miR-145-5p reduced GC cell viability and colony formation and induced GC cell apoptosis, whereas overexpression of ARF6 attenuated the effects of overexpression of miR-145-5p on GC cell viability, colony formation and apoptosis.
CONCLUSIONS
KCNQ1OT1 can promote GC progression through the miR-145-5p/ARF6 axis. KCNQ1OT1 may serve as a therapeutic target and a diagnostic biomarker of GC.
背景
长链非编码 RNA KCNQ1 反义链/反义转录本 1 基因（KCNQ1OT1）已被报道参与多种类型的人类癌症的进展，而其在胃癌（GC）中的作用尚不清楚。本研究旨在探讨 KCNQ1OT1 在 GC 中的作用。
方法
共收集 25 例 GC 组织和相邻正常组织。采用实时定量逆转录聚合酶链反应或 Western blot 检测 GC 组织和细胞系中 KCNQ1OT1、miR-145-5p 和 ARF6 的表达。通过生物信息学分析和双荧光素酶报告基因检测确定 KCNQ1OT1 与 miR-145-5p 或 miR-145-5p 与 ARF6 之间的关系。通过 KCNQ1OT1 和 miR-145-5p 的功能获得和缺失实验来证实它们在 GC 细胞中的作用。用转染 sh-KCNQ1OT1 或空载体的 BGC803 肿瘤细胞建立异种移植肿瘤模型，以确定 LINC01089 在体内的作用。
结果
KCNQ1OT1 在 GC 组织和细胞中的表达水平明显升高。敲低 KCNQ1OT1 抑制 GC 肿瘤生长，降低 GC 细胞活力和集落形成，并诱导 GC 细胞凋亡。GC 细胞中 miR-145-5p 的表达水平显著降低，与 GC 肿瘤中 KCNQ1OT1 的表达相关。此外，KCNQ1OT1 可直接与 miR-145-5p 结合，miR-145-5p 靶向 ARF6。敲低 KCNQ1OT1 增加了 miR-145-5p 的表达水平。抑制 miR-145-5p 增加了 KCNQ1OT1 的表达水平，并减弱了敲低 KCNQ1OT1 对 GC 细胞活力、增殖和凋亡的影响。此外，过表达 miR-145-5p 降低 GC 细胞活力和集落形成并诱导 GC 细胞凋亡，而过表达 ARF6 则减弱了过表达 miR-145-5p 对 GC 细胞活力、集落形成和凋亡的影响。
结论
KCNQ1OT1 可通过 miR-145-5p/ARF6 轴促进 GC 进展。KCNQ1OT1 可能作为 GC 的治疗靶点和诊断生物标志物。
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