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长链非编码 RNA NEAT1 通过海绵吸附 miR-365 调控 FGF9 促进卵巢癌细胞增殖和共孵育人脐静脉内皮细胞血管生成：一项实验研究。

LncRNA NEAT1 promotes proliferation of ovarian cancer cells and angiogenesis of co-incubated human umbilical vein endothelial cells by regulating FGF9 through sponging miR-365: An experimental study.

机构信息

Department of Obstetrics and Gynecology, West China Second University Hospital, Sichuan University; The Key Laboratory of Birth Defects and Related Diseases of Women and Children (Sichuan University), Ministry of Education, Sichuan, China.

出版信息

Medicine (Baltimore). 2021 Jan 22;100(3):e23423. doi: 10.1097/MD.0000000000023423.

DOI:10.1097/MD.0000000000023423
PMID:33545926
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7837846/
Abstract

OBJECTIVE

To uncover the function of lncRNA NEAT1 in ovarian cancer (OC) cells and its mechanism.

METHODS

The expression patterns of lncRNA NEAT1 and FGF9 in human OC cells and human ovarian epithelial cells was determined. OC cells were transfected with sh-NEAT1, pcDNA3.1-NEAT1, miR-365 mimic, miR-365 inhibitor or pcDNA3.1-NEAT1 + sh-NEAT1 before cell proliferation rate and cell clone formation rate were measured. After the transfected OC cells were co-cultivated with human umbilical vein endothelial cells (HUVECs), Matrigel angiogenesis assay tested angiogenesis of HUVECs; qRT-PCR and Western blot tested the expressions of vascular endothelial growth factor (VEGF), angiogenin 1 (Ang-1) and matrix metalloproteinase 2 (MMP2). Dual-luciferase reporter assay determined the targeted binding of NEAT1 and FGF9 to miR-365.

RESULTS

LncRNA NEAT1 and FGF9 are over-expressed in OC cells. Knockdown of NEAT1 or FGF9, or over-expression of miR-365 results in decreased proliferation rate and cell clones as well as inhibited angiogenesis and down-regulated expressions of VEGF, Ang-1 and MMP2. Over-expression of NEAT1 or knockdown of miR-365 can reverse the effect caused by FGF9 knockdown. NEAT1 can down-regulate the expression of miR-365 while up-regulating that of FGF9. Dual-luciferase reporter assay determined that NEAT1 competes with FGF9 for binding to miR-365.

CONCLUSION

LncRNA NEAT1 up-regulates FGF9 by sponging miR-365, thus promoting OC cell proliferation and angiogenesis of HUVECs.

摘要

目的

揭示长链非编码 RNA NEAT1 在卵巢癌（OC）细胞中的功能及其作用机制。

方法

检测长链非编码 RNA NEAT1 和 FGF9 在人 OC 细胞和人卵巢上皮细胞中的表达模式。转染 OC 细胞 sh-NEAT1、pcDNA3.1-NEAT1、miR-365 模拟物、miR-365 抑制剂或 pcDNA3.1-NEAT1+sh-NEAT1 后，检测细胞增殖率和细胞克隆形成率。转染的 OC 细胞与人脐静脉内皮细胞（HUVEC）共培养后，Matrigel 血管生成实验检测 HUVEC 的血管生成能力；qRT-PCR 和 Western blot 检测血管内皮生长因子（VEGF）、血管生成素 1（Ang-1）和基质金属蛋白酶 2（MMP2）的表达。双荧光素酶报告基因实验检测 NEAT1 和 FGF9 与 miR-365 的靶向结合。

结果

长链非编码 RNA NEAT1 和 FGF9 在 OC 细胞中高表达。敲低 NEAT1 或 FGF9，或过表达 miR-365，均可降低增殖率和细胞克隆数，抑制血管生成，并下调 VEGF、Ang-1 和 MMP2 的表达。过表达 NEAT1 或敲低 miR-365 可逆转 FGF9 敲低的作用。NEAT1 可下调 miR-365 的表达，同时上调 FGF9 的表达。双荧光素酶报告基因实验确定 NEAT1 可与 FGF9 竞争与 miR-365 结合。

结论

长链非编码 RNA NEAT1 通过海绵吸附 miR-365 上调 FGF9，从而促进 OC 细胞增殖和 HUVEC 血管生成。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/9af1e7a102d0/medi-100-e23423-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/1e9629f8a979/medi-100-e23423-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/079ecc08913a/medi-100-e23423-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/388993d08f8b/medi-100-e23423-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/2214f3f5ae1c/medi-100-e23423-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/9af1e7a102d0/medi-100-e23423-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/1e9629f8a979/medi-100-e23423-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/079ecc08913a/medi-100-e23423-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/388993d08f8b/medi-100-e23423-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/2214f3f5ae1c/medi-100-e23423-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/457c/7837846/9af1e7a102d0/medi-100-e23423-g005.jpg

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