绵羊miR-200b对卵泡颗粒细胞周期和凋亡的影响

doi:10.11843/j.issn.0366-6964.2021.012.014

Abstract

Abstract: The aim of study was to reveal the effect of ovine miR-200b on the follicular granulosa cells. The predicted target genes of miR-200b by 3 online tools (TargetScan, miRTarBase and miRDB) were analyzed with KOBAS for GO and KEGG pathway enrichment. The isolated and cultured sheep follicular granulosa cells were divided into 4 groups transfected with miR-200b mimic, mimic NC, miR-200b inhibitor and inhibitor NC,respectively, with 6 replicates in each group. CCK8 was used to detect the proliferation of granulosa cells at 24, 48 and 72 h after transfection. The expression level of miR-200b, CDK4, CDK6, CCND1, CCND2, Bax and Bcl-2 genes were measured by fluorescence quantitative PCR. A total of 25 common target genes of miR-200b were predicted by 3 online tools. These predicted target genes were enriched in cell proliferation and differentiation, cell cycle and reproductive development by GO and KEGG enrichment analysis. The survival rate of granulosa cell with time was a "V" decreased trendency and reached the lowest at 48 h (P<0.01) after transfection. There were no significant difference in miR-200b expression (P>0.05) between miR-200b inhibitor and inhibitor NC groups. Compared with mimic NC group, miR-200b mimic group significantly increased the expression of miR-200b (P<0.001), significantly down-regulated the expression of CDK4 (P<0.01), CDK6 (P<0.001), CCND1 (P<0.001), CCND2 (P<0.05) and Bcl-2 (P<0.01), while the expression of Bax did not significantly change (P>0.05), and extremely significantly decreased the ratio of Bcl-2 to Bax (P<0.001). In summary, miR-200b can inhibit the cell cycle and proliferation, and promote the apoptosis of follicular granulosa cells in sheep.

Key words: miR-200b, cell cycle, apoptosis, granulosa cell, sheep

CLC Number:

S826.3

SONG Pengyan, XI Jianzhong, ZHANG Zhenhong, FU Qiang, YUE Qiaoxian, ZHANG Peiying, ZHOU Rongyan. Effects of Ovine miR-200b on Cell Cycle and Apoptosis of Follicular Granulosa Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3471-3479.

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Effects of Ovine miR-200b on Cell Cycle and Apoptosis of Follicular Granulosa Cells

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