Abstract：Inhibitor of DNA binding 1 (ID1) is widely present in the body, and affects cell proliferation and apoptosis. To reveal the role of transcription factor ID1 in ovarian follicular granulosa cells of sheep (Ovis aries), the eukaryotic overexpression vector pcDNA3.1-ID1 was constructed and ID1 RNA interference fragment (siID1) was designed and synthesized. The expression level of ID1 gene was detected by qRT-PCR after 24 h of isolated and cultured ovarian follicular granulosa cells treated with follicle-stimulating hormone (FSH). The results showed that the expression of ID1 gene was extremely significantly decreased after FSH treatment (P<0.01). CCK-8 assay was used to detect the proliferation of granulosa cells within 5 d after transfection with pcDNA3.1, pcDNA3.1-ID1, siNC and siID1 in granulosa cells. After 48 h of transfection, JC-1 assay was used to detect the apoptosis of granulosa cells and the expression levels of proliferation and apoptosis genes were detected by qRT-PCR. The result showed that the survival rate of ID1 overexpressed granulosa cells was reduced, the green fluorescence of JC-1 was enhanced, the expression levels of proliferation-related genes cyclin dependent kinase 2 (CDK2)、cyclin dependent kinase 4 (CDK4) and cyclin D2 (CCND2) were significantly down-regulated (P<0.05), the expression of apoptosis-related gene B-cell lymphoma-2 (Bcl2) was significantly decreased (P<0.05), Bcl2-associated X protein (Bax) gene expression was significantly increased (P<0.05), and caspase-3 (Casp3) gene expression was extremely significantly increased (P<0.01). The higher survival rate of granulosa cells and the lower green fluorescence of JC-1 were found in cells with inhibited ID1 expression. The expression levels of proliferation-related genes CDK2 and CCND2 were significantly increased (P<0.01), CDK4 gene was significantly increased (P<0.05), the expression of Bcl2 was significantly increased (P<0.01), and the expression of Bax was significantly decreased (P<0.05). In summary, transcription factor ID1 inhibited the proliferation and promoted the apoptosis of sheep granulosa cells. This study provides a basis for exploring the biological functions of ID1 in follicular granulosa cells and revealing its role in ovarian follicle development.
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