Liu Yufang, Chen Yulin, Zhou Zuyang, He Xiaoyun, Tao Lin, Jiang Yanting, Lan Rong, Hong Qionghua, Chu Mingxing
Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, China.
College of Life Sciences and Food Engineering, Hebei University of Engineering, Handan, China.
Front Vet Sci. 2021 Nov 18;8:732440. doi: 10.3389/fvets.2021.732440. eCollection 2021.
Granulosa cell (GC) proliferation provides essential conditions for ovulation in animals. A previous study showed that plays a significant role in polycystic ovary syndrome. However, the modulation of in GCs remains unclear. Our previous integrated analysis of miRNA-mRNA revealed that the 3'-untranslated region of could be a target of chi-miR-324-3p. In this study, we used quantitative reverse transcription polymerase chain reaction (RT-qPCR) to investigate expression in ovarian tissues of high- and low-yielding goats. Furthermore, dual-fluorescent reporter vector experiments, Cell Counting Kit-8 (CCK-8) assay, and RT-qPCR were used to elucidate the regulatory pathway of chi-miR-324-3p- in GCs. The results revealed an opposite tendency between the expressions of chi-miR-324-3p and in the ovaries of high- and low-yielding goats. The CCK-8 assay indicated that chi-miR-324-3p overexpression significantly suppressed GC proliferation, whereas chi-miR-324-3p inhibition promoted GC proliferation. In addition, the expressions of GC proliferation markers , and showed the same tendency. The dual-fluorescent reporter assay revealed that chi-miR-324-3p directly targeted , and the RT-qPCR results revealed that expression was inhibited by chi-miR-324-3p. In summary, chi-miR-324-3p inhibited the proliferation of GCs by targeting .
颗粒细胞（GC）增殖为动物排卵提供了必要条件。先前的一项研究表明，[此处原文缺失某物质，未完整给出，翻译时按原文照写]在多囊卵巢综合征中起重要作用。然而，[此处原文缺失某物质，未完整给出，翻译时按原文照写]在颗粒细胞中的调节作用仍不清楚。我们之前对miRNA-mRNA的综合分析表明，[此处原文缺失某物质，未完整给出，翻译时按原文照写]的3'-非翻译区可能是chi-miR-324-3p的靶标。在本研究中，我们使用定量逆转录聚合酶链反应（RT-qPCR）来研究高产和低产山羊卵巢组织中[此处原文缺失某物质，未完整给出，翻译时按原文照写]的表达。此外，还使用了双荧光报告载体实验、细胞计数试剂盒-8（CCK-8）检测和RT-qPCR来阐明chi-miR-324-3p在颗粒细胞中的调控途径。结果显示，高产和低产山羊卵巢中chi-miR-324-3p和[此处原文缺失某物质，未完整给出，翻译时按原文照写]的表达呈相反趋势。CCK-8检测表明，chi-miR-324-3p过表达显著抑制颗粒细胞增殖，而抑制chi-miR-324-3p则促进颗粒细胞增殖。此外，颗粒细胞增殖标志物[此处原文缺失某物质，未完整给出，翻译时按原文照写]、[此处原文缺失某物质，未完整给出，翻译时按原文照写]和[此处原文缺失某物质，未完整给出，翻译时按原文照写]的表达也呈现相同趋势。双荧光报告检测表明，chi-miR-324-3p直接靶向[此处原文缺失某物质，未完整给出，翻译时按原文照写]，RT-qPCR结果显示[此处原文缺失某物质，未完整给出，翻译时按原文照写]的表达受到chi-miR-324-3p的抑制。总之，chi-miR-324-3p通过靶向[此处原文缺失某物质，未完整给出，翻译时按原文照写]抑制颗粒细胞的增殖。
