Lin28B overexpression decreases let-7b and let-7g levels and increases proliferation and estrogen secretion in Dolang sheep ovarian granulosa cells.

IF 1.6 3区 农林科学 Q2 AGRICULTURE, DAIRY & ANIMAL SCIENCE
Zhiyuan Sui, Yongjie Zhang, Zhishuai Zhang, Chenguang Wang, Xiaojun Li, Feng Xing
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引用次数: 0

Abstract

Although ovine puberty initiation has been previously studied, the mechanism by which the RNA-binding protein Lin28B affects this process has not been investigated. The present study aimed to investigate the effects of Lin28B overexpression on let-7b, let-7g, cell proliferation, and estrogen secretion in Dolang sheep ovine ovarian granulosa cells. In this study, a Lin28B vector was constructed and transfected into ovarian granulosa cells using liposomes. After 24, 48, and 72 h of overexpression, quantitative real-time PCR (qRT-PCR) was used for measuring let-7b and let-7g microRNA (miRNA) levels, and estrogen secretion was measured using the enzyme-linked immunosorbent assay (ELISA). A CCK-8 (Cell Counting Kit-8) kit was used for evaluating cell viability and proliferation in response to Lin28B overexpression at 24 h. The results showed that the expression of let-7b and let-7g decreased significantly after Lin28B overexpression, and the difference was consistent over different periods. The result of ELISA showed that estradiol (E2) levels significantly increased following Lin28B overexpression. Additionally, Lin28B overexpression significantly increased the cell viability and proliferation. Therefore, the Lin28B-let-7 family axis may play a key role in the initiation of female ovine puberty.

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Lin28B过表达降低了多郎羊卵巢颗粒细胞let-7b和let-7g水平,增加了卵巢颗粒细胞的增殖和雌激素分泌。
虽然以前已经研究过羊的青春期启动,但rna结合蛋白Lin28B影响这一过程的机制尚未被研究。本研究旨在探讨Lin28B过表达对多郎羊羊卵巢颗粒细胞let-7b、let-7g、细胞增殖及雌激素分泌的影响。本研究构建了Lin28B载体,利用脂质体将其转染到卵巢颗粒细胞中。过表达24、48和72 h后,采用实时荧光定量PCR (qRT-PCR)检测let-7b和let-7g microRNA (miRNA)水平,酶联免疫吸附试验(ELISA)检测雌激素分泌。使用CCK-8(细胞计数试剂盒-8)试剂盒评估细胞活力和增殖对Lin28B过表达24 h的响应。结果表明,过表达Lin28B后,let-7b和let-7g的表达均显著降低,且不同时期差异一致。ELISA结果显示,Lin28B过表达后雌二醇(E2)水平显著升高。此外,Lin28B过表达显著提高了细胞活力和增殖能力。因此,Lin28B-let-7家族轴可能在母羊青春期的开始中起关键作用。
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来源期刊
Archiv Fur Tierzucht-Archives of Animal Breeding
Archiv Fur Tierzucht-Archives of Animal Breeding 农林科学-奶制品与动物科学
CiteScore
3.20
自引率
0.00%
发文量
41
审稿时长
18-36 weeks
期刊介绍: Archives Animal Breeding is an open-access journal publishing original research papers, short communications, brief reports, and reviews by international researchers on scientific progress in farm-animal biology. The journal includes publications in quantitative and molecular genetics, genetic diversity, animal husbandry and welfare, physiology, and reproduction of livestock. It addresses researchers, teachers, stakeholders of academic and educational institutions, as well as industrial and governmental organizations in the field of animal production.
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