Abstract
Cytochrome P450 aromatase 19A1 (CYP19A1) is a critical enzyme in estrogen synthesis. However, the effect of CYP19A1 on cell growth and hormone secretion of buffalo follicular granulosa cells (BFGCs) is poorly understood. The objective of this study was to assess the role of CYP19A1 in cell proliferation and hormone secretion of BFGCs by knocking down CYP19A1 mRNA expression. The mRNA expression level of CYP19A1 gene was knocked down in BFGCs using the siCYP19A1-296 fragment with the best interference efficiency of 72.63%, as affirmed by real-time quantitative PCR (qPCR) and cell morphology analysis. The CYP19A1 knockdown promoted the proliferation of BFGCs through upregulating the mRNA expression levels of six proliferation-related genes (CCND1, CCNE1, CCNB1, CDK2, CDKN1A, and CDKN1B). Moreover, CYP19A1 knockdown increased (P < 0.05) the concentrations of progesterone secretion (P4) in BFGCs through increasing the mRNA expression levels of three steroidogenic genes (HSD17B1, HSD17B7, and CYP17A1). Our data further found that the FSH could inhibit the mRNA expression level of CYP19A1 in BFGCs, while LH obtains the opposite effect. These findings showed that the CYP19A1 knockdown had a regulatory role in the hormone secretion and cell proliferation in BFGCs.
Highlights
With the action of reproductive hormones among the hypothalamic-pituitary-gonadal axis, follicular cells could promote the expression of steroid hormone synthase, guide the production of steroid hormones, and further regulate the growth and development of follicular
The buffalo follicular granulosa cells (BFGCs) treated by siCYP19A1-296 remained a flat elongated shape after 72 h, with a small number of cells began to enter the early apoptosis (Figure 1B)
The results indicated that the increases (P < 0.05) in the mRNA expression level of the studied genes were observed after Cytochrome P450 aromatase 19A1 (CYP19A1) silencing (Figure 1C)
Summary
With the action of reproductive hormones among the hypothalamic-pituitary-gonadal axis, follicular cells could promote the expression of steroid hormone synthase, guide the production of steroid hormones, and further regulate the growth and development of follicular. Estradiol (E2) produced by the interaction of steroid hormone synthase in follicular membrane cells and granulosa cells, which help in promoting the proliferation of granulosa cells and regulating the differentiation of follicular cells in the ovaries [1]. Cholesterol side-chain cleavage enzyme is commonly referred to as P450SCC, which is a member of the cytochrome P450 family enzyme. The cytochrome P450 family 11 subfamily A member 1 (CYP11A1) converts cholesterol from follicular theca cells into progesterone (P5), which is the first and rate-limiting enzymatic step in the production of all steroid hormones [3].
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