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Wenzhi REN






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Journal of Zhejiang University SCIENCE B 2022 Vol.23 No.6 P.502-514


LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells

Author(s):  Weidi ZHANG, Wenzhi REN, Dongxu HAN, Guokun ZHAO, Haoqi WANG, Haixiang GUO, Yi ZHENG, Zhonghao JI, Wei GAO, Bao YUAN

Affiliation(s):  Department of Laboratory Animals, College of Animal Sciences, Jilin University, Changchun 130062, China; more

Corresponding email(s):   gaowei81@jlu.edu.cn, yuan_bao@jlu.edu.cn

Key Words:  Long noncoding RNA (lncRNA), MicroRNA (miRNA), Competitive endogenous RNA (ceRNA), Follicle-stimulating hormone (FSH), Mothers against decapentaplegic homolog 2/3 (Smad2/3)

Weidi ZHANG, Wenzhi REN, Dongxu HAN, Guokun ZHAO, Haoqi WANG, Haixiang GUO, Yi ZHENG, Zhonghao JI, Wei GAO, Bao YUAN. LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells[J]. Journal of Zhejiang University Science B, 2022, 23(6): 502-514.

@article{title="LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells",
author="Weidi ZHANG, Wenzhi REN, Dongxu HAN, Guokun ZHAO, Haoqi WANG, Haixiang GUO, Yi ZHENG, Zhonghao JI, Wei GAO, Bao YUAN",
journal="Journal of Zhejiang University Science B",
publisher="Zhejiang University Press & Springer",

%0 Journal Article
%T LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells
%A Weidi ZHANG
%A Wenzhi REN
%A Dongxu HAN
%A Guokun ZHAO
%A Haoqi WANG
%A Haixiang GUO
%A Zhonghao JI
%A Wei GAO
%J Journal of Zhejiang University SCIENCE B
%V 23
%N 6
%P 502-514
%@ 1673-1581
%D 2022
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.B2101052

T1 - LncRNA-m18as1 competitively binds with miR-18a-5p to regulate follicle-stimulating hormone secretion through the Smad2/3 pathway in rat primary pituitary cells
A1 - Weidi ZHANG
A1 - Wenzhi REN
A1 - Dongxu HAN
A1 - Guokun ZHAO
A1 - Haoqi WANG
A1 - Haixiang GUO
A1 - Zhonghao JI
A1 - Wei GAO
A1 - Bao YUAN
J0 - Journal of Zhejiang University Science B
VL - 23
IS - 6
SP - 502
EP - 514
%@ 1673-1581
Y1 - 2022
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.B2101052

Long noncoding RNAs (lncRNAs) are expressed in different species and different tissues, and perform different functions, but little is known about their involvement in the synthesis or secretion of follicle-stimulating hormone (FSH). In general, we have revealed lncRNA‍‒‍microRNA (miRNA)‍‒‍‍messenger RNA (mRNA) interactions that may play important roles in rat primary pituitary cells. In this study, a new lncRNA was identified for the first time. First, we analyzed the gene expression of lncRNA-m18as1 in different tissues and different stages by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and observed the localization of lncRNA-m18as1 with fluorescence in situ hybridization, which indicated that this lncRNA was distributed mainly in the cytoplasm. Next, we used RT-qPCR and enzyme-linked immunosorbent assay (ELISA) to analyze the regulation of FSH synthesis and secretion after overexpression or knockdown of lncRNA-m18as1 and found that lncRNA-m18as1 was positively correlated with FSH synthesis and secretion. In addition, mothers against decapentaplegic homolog 2 (Smad2) was highly expressed in our sequencing results. We also screened miR-18a-5p from our sequencing results as a miRNA that may bind to lncRNA-m18as1 and Smad2. We used RNA immunoprecipitation-qPCR (RIP-qPCR) and/or dual luciferase assays to confirm that lncRNA-m18as1 interacted with miR-18a-5p and miR-18a-5p interacted with Smad2. Fluorescence in situ hybridization (FISH) showed that lncRNA-m18as1 and miR-18a-5p were localized mainly in the cytoplasm. Finally, we determined the relationship among lncRNA-m18as1, miR-18a-5p, and the Smad2/3 pathway. Overall, we found that lncRNA-m18as1 acts as a molecular sponge of miR-18a-5p to regulate the synthesis and secretion of FSH through the Smad2/3 pathway.


方法:我们通过逆转录定量聚合酶链反应(RT-qPCR)筛选出了一个新的lncRNA,并根据功能将其最终命名为lncRNA-m18as1。经过敲降或过表达lncRNA-m18as1后,我们采用RT-qPCR与酶联免疫吸附剂测定(ELISA)分析了lncRNA-m18as1对FshβmRNA以及FSH分泌的调控作用。我们预测并确定了lncRNA-m18as1发挥作用的lncRNAm18as1/miR-18a-5p/Smad2轴。我们使用RNA结合蛋白免疫沉淀测定-逆转录定量聚合酶链反应(RIP-qPCR)和/或双荧光素酶报告分析方法分析了miR-18a-5p与lncRNA-m18as1、Smad2的靶向关系。此外,我们使用RT-qPCR和蛋白质印迹法(western blot)分析了lncRNA-m18as1和miR-18a-5p在轴中对向下游因子的调控作用,同时通过荧光原位杂交技术(FISH)观察了lncRNA-m18as1以及miR-18a-5p在细胞核与细胞质中的分布。


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