scholarly journals Activation of Steroidogenesis, Anti-Apoptotic Activity, and Proliferation in Porcine Granulosa Cells by RUNX1 Is Negatively Regulated by H3K27me3 Transcriptional Repression

Genes ◽  
2020 ◽  
Vol 11 (5) ◽  
pp. 495
Author(s):  
Yuyi Zhong ◽  
Liying Li ◽  
Yingting He ◽  
Bo He ◽  
Zhonghui Li ◽  
...  
Keyword(s):  
Transcription Factor ◽  
Granulosa Cells ◽  
Cell Apoptosis ◽  
Transcriptional Repression ◽  
Epigenetic Modification ◽  
Follicular Development ◽  
Hormone Synthesis ◽  
Porcine Granulosa Cells ◽  
Related Transcription Factor ◽  
Apoptotic Activity

H3K27me3 is an epigenetic modification that results in the repression of gene transcription. The transcription factor RUNX1 (the runt-related transcription factor 1) influences granulosa cells’ growth and ovulation. This research uses ELISA, flow cytometry, EDU, ChIP-PCR, WB and qPCR to investigate steroidogenesis, cell apoptosis, and the proliferation effect of RUNX1 in porcine granulosa cells (pGCs) as regulated by H3K27me3. Decreased H3K27me3 stimulates the expression of steroidogenesis-related genes, including CYP11A1, PTGS2, and STAR, as well as prostaglandin. H3K27me3 transcriptionally represses RUNX1 here, whereas RUNX1 acts as an activator of FSHR, CYP11A1, and CYP19A1, promoting the production of androgen, estrogen, and prostaglandin, as well as increasing anti-apoptotic and cell proliferation activity, but decreasing progesterone. Both the complementary recovery of the H3K27me3 antagonist with the siRUNX1 signal, and the H3K27me3 agonist with the RUNX1 signal to maintain RUNX1 lead to the activation of CYP19A1, ER1, HSD17β4, and STAR here. Androgen and prostaglandin are significantly repressed but progesterone is markedly increased with the antagonist and siRUNX1. Prostaglandin is significantly promoted with the agonist and RUNX1. Furthermore, H3K27me3-RUNX1 affects the anti-apoptotic activity and stimulation of proliferation in pGCs. The present work verifies the transcriptional suppression of RUNX1 by H3K27me3 during antral follicular development and maturation, which determines the levels of hormone synthesis and cell apoptosis and proliferation in the pGC microenvironment.

scholarly journals Effects of NRF1 on steroidogenesis and apoptosis in goat luteinized granulosa cells

Reproduction ◽  
2017 ◽  
Vol 154 (2) ◽  
pp. 111-122 ◽  
Author(s):  
Guo-Min Zhang ◽  
Ming-Tian Deng ◽  
Zhi-Hai Lei ◽  
Yong-Jie Wan ◽  
Hai-Tao Nie ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Cell Apoptosis ◽  
Follicular Development ◽  
Redox Balance ◽  
Stress System ◽  
Mtdna Copy Number ◽  
Aberrant Expression ◽  
Hormone Synthesis ◽  
Nuclear Respiratory Factor ◽  
Mitochondrial Transcription Factor

During goat follicular development, abnormal expression of nuclear respiratory factor 1 (NRF1) in granulosa cells may drive follicular atresia with unknown regulatory mechanisms. In this study, we investigated the effects of NRF1 on steroidogenesis and cell apoptosis by overexpressing or silencing it in goat luteinized granulosa cells (LGCs). Results showed that knockdown of NRF1 expression significantly inhibited the expression of STAR and CYP19A1, which are involved in sex steroid hormones synthesis, and led to lower estrogen levels. Knockdown of NRF1 resulted in an increased percentage of apoptosis, probably due to the release of cytochrome c from mitochondria, accompanied by upregulating mRNA and protein levels of apoptosis-related markers BAX, caspase 3 and caspase 9. These data indicate that NRF1 might be related with steroidogenesis and cell apoptosis. Furthermore, NRF1 silence reduced mitochondrial transcription factor A (TFAM) transcription activity, mtDNA copy number and ATP level. Simultaneously, knockdown of NRF1 suppressed the transcription and translation levels of SOD, GPx and CAT, decreased glutathione level and increased 8-OHdG level. However, the overexpression of NRF1 in LGCs or gain of TFAM in NRF1 silenced LGCs increased the expression of genes involved in mitochondrial function and biogenesis, and elevated the antioxidant stress system and steroids synthesis. Taken together, aberrant expression of NRF1 could induce mitochondrial dysfunction and disturb the cellular redox balance, which lead to disturbance of steroid hormone synthesis, and trigger LGC apoptosis through the mitochondria-dependent pathway. These findings will be helpful for understanding the role of NRF1 in goat ovarian follicular development and atresia.

scholarly journals Runt-Related Transcription Factor 1 Regulates Luteinized Hormone-Induced Prostaglandin-Endoperoxide Synthase 2 Expression in Rat Periovulatory Granulosa Cells

Endocrinology ◽  
2009 ◽  
Vol 150 (7) ◽  
pp. 3291-3300 ◽  
Author(s):  
Jing Liu ◽  
Eun-Sil Park ◽  
Misung Jo
Keyword(s):  
Gene Expression ◽  
Transcription Factor ◽  
Granulosa Cells ◽  
Promoter Region ◽  
Transcriptional Activity ◽  
Binding Motifs ◽  
Related Transcription Factor ◽  
Prostaglandin Endoperoxide Synthase ◽  
Transcription Factor 1 ◽  
Runx1 Expression

Runt-related transcription factor 1 (RUNX1), a transcription factor, is transiently induced by the LH surge and regulates gene expression in periovulatory granulosa cells. Potential binding sites for RUNX are present in the 5′-flanking region of the Ptgs2 (prostaglandin-endoperoxide synthase 2) gene. Periovulatory Ptgs2 expression is essential for ovulation. In the present study, we investigated the role of RUNX1 in mediating the LH-induced expression of Ptgs2 in periovulatory granulosa cells. We first determined whether the suppression of Runx1 expression or activity affects Ptgs2 expression using cultured preovulatory granulosa cells isolated from immature rat ovaries primed with pregnant mare serum gonadotropin for 48 h. Knockdown of human chorionic gonadotropin-induced Runx1 expression by small interfering RNA or inhibition of endogenous RUNX activities by dominant-negative RUNX decreased human chorionic gonadotropin or agonist-stimulated Ptgs2 expression and transcriptional activity of Ptgs2 promoter reporter constructs. Results from chromatin immunoprecipitation assays revealed in vivo binding of endogenous RUNX1 to the Ptgs2 promoter region in rat periovulatory granulosa cells. Direct binding of RUNX1 to two RUNX-binding motifs in the Ptgs2 promoter region was confirmed by EMSA. The mutation of these two binding motifs resulted in decreased transcriptional activity of Ptgs2 promoter reporter constructs in preovulatory granulosa cells. Taken together, these findings provide experimental evidence that the LH-dependent induction of Ptgs2 expression results, in part, from RUNX1-mediated transactivation of the Ptgs2 promoter. The results of the present study assign potential significance for LH-induced RUNX1 in the ovulatory process via regulating Ptgs2 gene expression.

scholarly journals MiR-26a promotes apoptosis of porcine granulosa cells by targeting the 3β-hydroxysteroid-Δ24-reductase gene

2020 ◽  
Vol 33 (4) ◽  
pp. 547-555 ◽  
Author(s):  
Xiaodong Zhang ◽  
Qiangqiang Tao ◽  
Jinnan Shang ◽  
Yiliang Xu ◽  
Liang Zhang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Granulosa Cells ◽  
Cell Apoptosis ◽  
Binding Sites ◽  
Follicular Development ◽  
Reproductive Capacity ◽  
Regulatory Relationship ◽  
Reductase Gene ◽  
Mrna And Protein Expression

Objective: Apoptosis of ovarian granulosa cells (GCs) affects mammalian follicular development and fecundity. This study aimed to explore the regulatory relationship between microRNA-26a (miR-26a) and the 3β-hydroxysteroid-Δ24-reductase gene (<i>DHCR24</i>) gene in porcine follicular granular cells (pGCs), and to provide empirical data for the development of methods to improve the reproductive capacity of pigs.Methods: The pGCs were transfected with miR-26a mimic, miR-26a inhibitor and <i>DHCR24</i>-siRNA <i>in vitro</i>. The cell apoptosis rate of pGCs was detected by the flow cytometry. The secretion levels of estradiol (E2) and progesterone (P) in pGCs were detected by enzymelinked immunosorbent assay. Double luciferase validation system was used to detect the binding sites between miR-26a and <i>DHCR24</i> 3′-UTR region. Qualitative real-time polymerase chain reaction and Western blotting were used to verify the <i>DHCR24</i> mRNA and protein expression in pGCs, respectively, after transfecting with miR-26a mimic and miR-26a inhibitor.Results: Results showed that enhancement of miR-26a promoted apoptosis, and inhibited E2 and P secretion in pGCs. Meanwhile, inhibition of <i>DHCR24</i> also upregulated the Caspase-3 expression, reduced the BCL-2 expression, promoted pGCs apoptosis, and inhibited E2 and P secretion in pGCs. There were the binding sites of miR-26a located within <i>DHCR24</i> 3′-UTR. Up-regulation of miR-26a inhibited <i>DHCR24</i> mRNA and protein expression in pGCs.Conclusion: This study demonstrates that miR-26a can promote cell apoptosis and inhibit E2 and P secretion by inhibiting the expression of <i>DHCR24</i> in pGCs.

MiR-31 and miR-143 affect steroid hormone synthesis and inhibit cell apoptosis in bovine granulosa cells through FSHR

2019 ◽  
Vol 123 ◽  
pp. 45-53 ◽  
Author(s):  
Zhe Zhang ◽  
Cheng-Zhen Chen ◽  
Ming-Qiang Xu ◽  
Li-Qun Zhang ◽  
Jian-Bo Liu ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Cell Apoptosis ◽  
Steroid Hormone ◽  
Hormone Synthesis

MiR-126-3p promotes the cell proliferation and inhibits the cell apoptosis by targeting TSC1 in the porcine granulosa cells

2018 ◽  
Vol 54 (10) ◽  
pp. 715-724 ◽  
Author(s):  
Xiaolong Yuan ◽  
Xi Deng ◽  
Xiaofeng Zhou ◽  
Ailing Zhang ◽  
Yan Xing ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Granulosa Cells ◽  
Cell Apoptosis ◽  
Porcine Granulosa Cells

Bmal1 interference impairs hormone synthesis and promotes apoptosis in porcine granulosa cells

2017 ◽  
Vol 99 ◽  
pp. 63-68 ◽  
Author(s):  
Wusu Wang ◽  
Lin Yin ◽  
Long Bai ◽  
Guangjun Ma ◽  
Cunzhen Zhao ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Hormone Synthesis ◽  
Porcine Granulosa Cells

scholarly journals Micro-RNA378 (miR-378) Regulates Ovarian Estradiol Production by Targeting Aromatase

Endocrinology ◽  
2011 ◽  
Vol 152 (10) ◽  
pp. 3941-3951 ◽  
Author(s):  
Shengyu Xu ◽  
Katja Linher-Melville ◽  
Burton B. Yang ◽  
De Wu ◽  
Julang Li
Keyword(s):  
Granulosa Cells ◽  
Follicular Development ◽  
Micro Rna ◽  
Transcriptional Level ◽  
Reproductive Disorders ◽  
Aromatase Expression ◽  
Directed Mutation ◽  
Porcine Granulosa Cells ◽  
Posttranscriptional Level

Estradiol is a steroid hormone that not only plays an important role in ovarian follicular development but also is associated with many reproductive disorders. Owing to the importance of aromatase in the production of estradiol, the regulation of aromatase gene expression at the transcriptional level has been an extensive area of study for over two decades. However, its regulation at the posttranscriptional level has remained unclear. Here, we show that micro-RNA378 (miR-378) is spatiotemporally expressed in porcine granulosa cells, the cells that generate estradiol in the ovary during follicular development, in an inverse manner compared with the expression of aromatase. In vitro overexpression and inhibition experiments revealed that aromatase expression, and therefore estradiol production, by granulosa cells, is posttranscriptionally down-regulated by miR-378. Furthermore, site-directed mutation studies identified two binding sites in the 3′-untranslated region (3′-UTR) of the aromatase coding sequence that are critical for the action of miR-378. Interestingly, overexpression of the aromatase 3′-UTR enhanced aromatase expression at the protein level in granulosa cells, possibly mediated by the binding of miR-378 within this region, thereby reducing the binding of this micro-RNA to the endogenous aromatase 3′-UTR.

scholarly journals Effects of nitric oxide on steroidogenesis in porcine granulosa cells during different stages of follicular development

2001 ◽  
pp. 303-308 ◽  
Author(s):  
M Masuda ◽  
T Kubota ◽  
T Aso
Keyword(s):  
Nitric Oxide ◽  
Granulosa Cells ◽  
Follicular Development ◽  
Aromatase Activity ◽  
Follicular Growth ◽  
No Donor ◽  
Porcine Granulosa Cells ◽  
Basal Release ◽  
Independent Process ◽  
Design And Methods

BACKGROUND: We have previously demonstrated that nitric oxide (NO) inhibits steroidogenesis via a cGMP-independent process, by inhibiting P450 aromatase activity in porcine granulosa cells (PGCs) derived from medium-sized (3--5 mm) ovarian follicles (M-PGC). OBJECTIVE: To determine whether the NO/NO synthase (NOS) system exerts any significant effects on steroidogenesis in PGCs derived from small follicles (<3 mm) (S-PGC) in comparison with those derived from medium follicles. DESIGN AND METHODS: PGCs, namely S-PGC and M-PGC, were incubated with the NO donor, NOC18, and a competitive blocker of NOS, N(3)-monomethyl-l-arginine (LNMMA), either alone or in the presence of FSH (200 ng/ml) or hCG (5 IU/ml). RESULTS: NOC18 significantly (P<0.01--0.001) suppressed basal (unstimulated) and gonadotropin-stimulated estradiol (E2) release from both S-PGC and M-PGC in a 2-h culture. NOC18 significantly (P<0.01--0.001) decreased basal and gonadotropin-stimulated progesterone release from S-PGC, but not from M-PGC. In addition, NOC18 significantly (P<0.05--0.001) inhibited aromatase activity in S-PGC. LNMMA had a significantly (P<0.01--0.001) stimulatory effect on the basal release of E2 and progesterone from M-PGC; however, it had no significant effect on basal steroidogenesis in S-PGC in a 24-h culture. In the presence of gonadotropin, LNMMA significantly (P<0.01--0.001) stimulated the release of E2 and progesterone from both S- and M-PGC, and this stimulatory effect was weaker in S-PGC than in M-PGC. These results demonstrate that NO inhibits E2 secretion by directly inhibiting the aromatase activity in S-PGC, as in M-PGC. It has been shown that the NO system suppresses the differentiation of S-PGC; however, the extent of suppression decreased with the progression of follicular growth. In addition, the activity of NOS in S-PGC was weaker than that in M-PGC. CONCLUSION: We strongly suggest that the NO/NOS system in PGC regulates steroidogenesis differently during different phase of follicular development.

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