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长链非编码 RNA TTN-AS1 作为 miR-15b-5p 的海绵体调节卵巢癌中 FBXW7 的表达。

LncRNA TTN-AS1 acts as sponge for miR-15b-5p to regulate FBXW7 expression in ovarian cancer.

机构信息

Department of Gynaecology and Obstetrics, China-Japan Union Hospital of Jilin University, Changchun, China.

出版信息

Biofactors. 2020 Jul;46(4):600-607. doi: 10.1002/biof.1622. Epub 2020 Feb 12.

Abstract

Emerging evidence showed that long noncoding RNA (lncRNA) plays crucial roles in regulating various cancer biological behaviors. Titin-antisense RNA1 (TTN-AS1) has been reported to have crucial roles in cancers but its role in ovarian cancer remains unknown. The levels of TTN-AS1, microNRA-15b-5p (miR-15b-5p), and F-box and WD repeat domain containing 7 (FBXW7) in ovarian cancer cells were measured by quantitative reverse-transcription PCR. Targets for TTN-AS1 and miR-15b-5p were predicted by bioinformatic tools, and validated by luciferase activity reporter assay. Cell proliferation, colony formation, and cell apoptosis were analyzed with cell counting kit-8 assay, colony formation assay, and flow cytometry. Correlation of TTN-AS1 and FBXW7 was analyzed at gene expression profiling interactive analysis. TTN-AS1 was found decreased expression in ovarian cancer tissues and cells. Dual-luciferase activity validated TTN-AS1 and FBXW7 shared binding site in miR-15b-5p. Functional assays showed TTN-AS1 overexpression inhibits ovarian cancer cell proliferation, colony formation but promotes apoptosis. Rescue experiments showed that knockdown of FBXW7 could partially counteracted the effects of TTN-AS1 overexpression on ovarian cancer cell behaviors. Our results indicated that the TTN-AS1/miR-15b-5p/FBXW7 axis identified in this work could help to identify treatment biomarkers for ovarian cancer.

摘要

中文译文

越来越多的证据表明,长链非编码 RNA(lncRNA)在调节各种癌症生物学行为中发挥着关键作用。肌联蛋白反义 RNA1(TTN-AS1)已被报道在癌症中具有重要作用,但它在卵巢癌中的作用尚不清楚。通过定量逆转录 PCR 测量卵巢癌细胞中的 TTN-AS1、microNRA-15b-5p(miR-15b-5p)和 F-box 和 WD 重复域包含 7(FBXW7)的水平。通过生物信息学工具预测 TTN-AS1 和 miR-15b-5p 的靶标,并通过荧光素酶活性报告基因测定验证。通过细胞计数试剂盒-8 测定、集落形成测定和流式细胞术分析细胞增殖、集落形成和细胞凋亡。在基因表达谱交互分析中分析 TTN-AS1 和 FBXW7 的相关性。在卵巢癌组织和细胞中发现 TTN-AS1 表达降低。双荧光素酶活性验证 TTN-AS1 和 FBXW7 共享 miR-15b-5p 的结合位点。功能测定表明,TTN-AS1 过表达抑制卵巢癌细胞增殖、集落形成,但促进凋亡。挽救实验表明,FBXW7 的敲低可以部分抵消 TTN-AS1 过表达对卵巢癌细胞行为的影响。我们的结果表明,本工作中鉴定的 TTN-AS1/miR-15b-5p/FBXW7 轴可能有助于识别卵巢癌的治疗生物标志物。

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