长链非编码 RNA AFAP1-AS1 通过调控 miR-107/PDK4 轴促进卵巢癌细胞的进展。

BACKGROUND

Abnormally expressed in various tumors, long non-coding RNAs (lncRNAs) feature prominently in tumor development, yet little is still known regarding the functional roles of lncRNA AFAP1 antisense RNA 1 (AFAP1-AS1) in ovarian cancer (OC).

METHODS

The relative expression levels of lncRNA AFAP1-AS1, microRNA (miR)-107 and pyruvate dehydrogenase kinase isozyme 4 (PDK4) mRNA were assessed by quantitative real-time PCR. PDK4, PCNA and cyclin D1 expression levels were determined using Western blot analysis. Bioinformatics analysis and dual-luciferase gene reporter assay were conducted for identifying and validating the binding sequences between AFAP1-AS1 and miR-107, as well as between miR-107 and PDK4. Cell counting kit-8 assay was employed for detecting cell proliferation. Cell migration and invasion abilities were examined using Transwell assays.

RESULTS

The present study revealed that AFAP1-AS1 expression was elevated in OC cells and tissues. AFAP1-AS1 expression and FIGO stage were positively correlated. AFAP1-AS1 knockdown repressed OC cell proliferation, migration and invasion. AFAP1-AS1 functioned as a sponge of miR-107, and miR-107 reversed the effects of AFAP1-AS1 on OC cells. It was validated that miR-107 was able to bind to PDK4, and AFAP1-AS1 regulated PDK4 expression by competitively binding with miR-107. Additionally, miR-107 modulated OC cell proliferation, migration and invasion via targeting PDK4.

CONCLUSIONS

LncRNA AFAP1-AS1 serves as a tumor driver in the pathogenesis of OC via the miR-107/PDK4 axis.

背景

长链非编码 RNA（lncRNA）在各种肿瘤中异常表达，在肿瘤发生发展中起重要作用，但 lncRNA AFAP1 反义 RNA 1（AFAP1-AS1）在卵巢癌（OC）中的功能作用仍知之甚少。

方法

采用实时定量 PCR 检测 lncRNA AFAP1-AS1、微小 RNA（miR）-107 和丙酮酸脱氢酶激酶同工酶 4（PDK4）mRNA 的相对表达水平。采用 Western blot 分析检测 PDK4、PCNA 和细胞周期蛋白 D1 的表达水平。通过生物信息学分析和双荧光素酶基因报告基因实验鉴定和验证 AFAP1-AS1 与 miR-107 以及 miR-107 与 PDK4 之间的结合序列。采用细胞计数试剂盒-8 检测细胞增殖。通过 Transwell 实验检测细胞迁移和侵袭能力。

结果

本研究表明，AFAP1-AS1 在 OC 细胞和组织中表达上调。AFAP1-AS1 表达与 FIGO 分期呈正相关。AFAP1-AS1 敲低抑制 OC 细胞增殖、迁移和侵袭。AFAP1-AS1 作为 miR-107 的海绵，miR-107 逆转了 AFAP1-AS1 对 OC 细胞的作用。验证了 miR-107 能够与 PDK4 结合，AFAP1-AS1 通过与 miR-107 竞争结合来调节 PDK4 的表达。此外，miR-107 通过靶向 PDK4 调节 OC 细胞的增殖、迁移和侵袭。