Department of Obstetrics and Gynecology, Minhang Hospital, Fudan University, Shanghai, China.
Department of Clinical Laboratory, Minhang Hospital, Fudan University, Shanghai, China.
Cell Prolif. 2020 Jul;53(7):e12823. doi: 10.1111/cpr.12823. Epub 2020 Jun 9.
Over the past years, growing attention has been paid to deciphering the pivotal role of long non-coding RNAs (lncRNAs) in regulating the occurrence and development of human malignancies, cervical cancer (CC) included. Nonetheless, the regulatory role of lncRNA BBOX1 antisense RNA 1 (BBOX1-AS1) has not been explored as yet.
The expression of BBOX1-AS1 was detected by reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR). Cell Counting Kit-8 (CCK-8), colony formation, TUNEL, Western blot, transwell and immunofluorescence assays testified the critical role of BBOX1-AS1 in CC. The relationship between RNAs (BBOX1-AS1, miR-361-3p, HOXC6 and HuR) was analysed by luciferase reporter, RNA Immunoprecipitation (RIP) and RNA pull-down assays.
BBOX1 antisense RNA 1 antisense RNA 1 was revealed to be highly expressed in CC. Decreased expression of BBOX1-AS1 had suppressive effects on CC cell growth and migration. Molecular mechanism assays verified that BBOX1-AS1 had negative interaction with miR-361-3p in CC. Additionally, homeobox C6 (HOXC6) was validated to be a downstream target of miR-361-3p in CC. Furthermore, ELAV-like RNA-binding protein 1, also known as HuR, was uncovered to be capable of regulating the mRNA stability of HOXC6 in CC. More importantly, rescue assays delineated that knockdown of HuR after overexpressing miR-361-3p could reverse BBOX1-AS1 upregulation-mediated effect on CC progression. Similarly, the function induced by BBOX1-AS1 upregulation on CC progression could be countervailed by HOXC6 depletion.
BBOX1 antisense RNA 1 facilitates CC progression by upregulating HOXC6 expression via miR-361-3p and HuR.
近年来，人们越来越关注长链非编码 RNA（lncRNA）在调控包括宫颈癌（CC）在内的人类恶性肿瘤发生和发展中的关键作用。然而，lncRNA BBOX1 反义 RNA 1（BBOX1-AS1）的调节作用尚未被探索。
采用逆转录实时定量聚合酶链反应（RT-qPCR）检测 BBOX1-AS1 的表达。细胞计数试剂盒-8（CCK-8）、集落形成、TUNEL、Western blot、transwell 和免疫荧光试验证实了 BBOX1-AS1 在 CC 中的关键作用。通过荧光素酶报告、RNA 免疫沉淀（RIP）和 RNA 下拉实验分析 RNA（BBOX1-AS1、miR-361-3p、HOXC6 和 HuR）之间的关系。
CC 中 BBOX1 反义 RNA 1 反义 RNA 1 表达水平较高。BBOX1-AS1 表达下调对 CC 细胞生长和迁移有抑制作用。分子机制试验证实，BBOX1-AS1 在 CC 中与 miR-361-3p 呈负相关。此外，在 CC 中验证了 homeobox C6（HOXC6）是 miR-361-3p 的下游靶点。进一步发现，ELAV 样 RNA 结合蛋白 1，又称 HuR，能够调节 CC 中 HOXC6 的 mRNA 稳定性。更重要的是，过表达 miR-361-3p 后敲低 HuR 可以逆转 BBOX1-AS1 上调对 CC 进展的影响。同样，BBOX1-AS1 上调对 CC 进展的作用可以通过 HOXC6 缺失来抵消。
BBOX1 反义 RNA 1 通过 miR-361-3p 和 HuR 上调 HOXC6 表达促进 CC 进展。
