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长链非编码 RNA MSTRG.5970.28 通过 miR-133a-3p/ANOS1 通路调控鹅卵泡颗粒细胞的增殖和凋亡。

Long non-coding RNA MSTRG.5970.28 regulates proliferation and apoptosis of goose follicle granulosa cells via the miR-133a-3p/ANOS1 pathway.

机构信息

College of Animal Science, Xinjiang Agricultural University, Urumqi, China.

College of Animal Science, Xinjiang Agricultural University, Urumqi, China.

出版信息

Poult Sci. 2023 Mar;102(3):102451. doi: 10.1016/j.psj.2022.102451. Epub 2022 Dec 25.

Abstract

The development of follicles in the ovaries is a critical determinant of poultry egg production. There are existing studies on the follicular development patterns in poultry, but the specific regulatory mechanisms still need further study. In a previous study, we identified long non-coding RNA (lncRNA) MSTRG.5970.28, anosmin 1 (ANOS1), and its predicted target miR-133a-3p that may be associated with goose ovary development. However, the function of MSTRG.5970.28 in goose granulosa cells and its regulatory mechanisms affecting granulosa cell proliferation and apoptosis have not been reported. In the present study, MSTRG.5970.28 and miR-133a-3p overexpression and interference vectors were constructed. Combined with reverse-transcription real-time quantitative PCR (RT-qPCR), a dual luciferase activity assay, Cell Counting Kit-8 (CCK-8), and flow cytometric analysis, we investigated the role of the MSTRG.5970.28-miR-133a-3p-ANOS1 axis in goose follicular granulosa cells and the associated regulatory mechanisms. MSTRG.5970.28 was found to be localized in the cytoplasm and its expression was influenced by reproductive hormones. The targeting relationship among MSTRG.5970.28, ANOS1, and miR-133a-3p were verified by a dual luciferase activity assay. CCK-8 and apoptosis assays showed that MSTRG.5970.28 inhibited the proliferation and promoted apoptosis of goose granulosa cells. The regulatory role of miR-133a-3p on granulosa cell proliferation and apoptosis was opposite to MSTRG.5970.28. We found that the proliferative and apoptotic effects of granulosa cells caused by MSTRG.5970.28 overexpression were attenuated by miR-133a-3p. MSTRG.5970.28 functions as a competitive endogenous RNA that regulates ANOS1 expression by sponging miR-133a-3p and thus exerts regulatory functions in granulosa cells. In sum, the present study identified lncRNA MSTRG.5970.28 as associated with goose ovary development, which affects the expression of ANOS1 by targeting miR-133a-3p, thereby influencing the proliferation and apoptosis of goose granulosa cells.

摘要

卵泡在卵巢中的发育是家禽产蛋的关键决定因素。目前已有关于家禽卵泡发育模式的研究,但具体的调节机制仍需要进一步研究。在之前的一项研究中,我们鉴定出长链非编码 RNA(lncRNA)MSTRG.5970.28、anosmin 1(ANOS1)及其预测的靶基因 miR-133a-3p 可能与鹅卵巢发育有关。然而,MSTRG.5970.28 在鹅颗粒细胞中的功能及其影响颗粒细胞增殖和凋亡的调节机制尚未报道。本研究构建了 MSTRG.5970.28 和 miR-133a-3p 的过表达和干扰载体。通过逆转录实时定量 PCR(RT-qPCR)、双荧光素酶活性测定、细胞计数试剂盒-8(CCK-8)和流式细胞术分析,研究了 MSTRG.5970.28-miR-133a-3p-ANOS1 轴在鹅卵泡颗粒细胞中的作用及其相关调节机制。发现 MSTRG.5970.28 定位于细胞质,其表达受生殖激素的影响。双荧光素酶活性测定验证了 MSTRG.5970.28 与 ANOS1 和 miR-133a-3p 之间的靶向关系。CCK-8 和凋亡实验表明,MSTRG.5970.28 抑制鹅颗粒细胞的增殖并促进其凋亡。miR-133a-3p 对颗粒细胞增殖和凋亡的调节作用与 MSTRG.5970.28 相反。我们发现,MSTRG.5970.28 过表达引起的颗粒细胞增殖和凋亡的调节作用被 miR-133a-3p 减弱。MSTRG.5970.28 作为一种竞争性内源性 RNA,通过海绵吸附 miR-133a-3p 调节 ANOS1 的表达,从而在颗粒细胞中发挥调节作用。总之,本研究鉴定出 lncRNA MSTRG.5970.28 与鹅卵巢发育有关,通过靶向 miR-133a-3p 影响 ANOS1 的表达,从而影响鹅颗粒细胞的增殖和凋亡。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/85c3/9841053/30b7dd293c59/gr1.jpg

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