Proliferation Of Granular Cells Research Articles

SEMA4G targeted by miR‐363‐5p regulates the proliferation of granulosa cells in Yunshang black goats

AbstractProliferation of granular cells (GCs) plays an important role in ovary development, providing energy and a microenvironment for oocyte ovulation. In this study, we explored the spatiotemporal expression of SEMA4G and its effects on the growth and development of goat GCs using primary GCs cultured in vitro as a model. The results showed that the expression level of SEMA4G was significantly higher in the ovaries of high‐fertility goats than in those of low‐fertility goats (p < 0.05). The mRNA and protein expression levels of the cell proliferation markers of GCs were significantly increased after the overexpression of SEMA4G in goat primary GCs. The EdU and CCK8 results showed that cell viability was elevated in goat GCs and that proliferation was promoted by an increase in the number of proliferating cells. The proliferation of goat GCs was significantly inhibited by SEMA4G inhibition (p < 0.05). The results of online miRNA and target gene prediction software and dual luciferase activity analysis confirmed that SEMA4G could bind to mi‐363‐5p and was one of its target genes. The RT‒qPCR results showed that the expression level of miR‐363‐5p was significantly lower in the ovaries of high‐fertility goats than in those of low‐fertility goats in contrast to the expression level of SEMA4G (p < 0.05). After the overexpression of miR‐363‐5p in goat GCs, the expression of SEMA4G was significantly suppressed (p < 0.05). Collectively, the results of this study could lay the foundation for exploring the molecular mechanisms by which SEMA4G and miR‐363‐5p regulate the growth and development of goat GCs and provide targets for breeding high‐fertility goats.

BMP4/SMAD8 signaling pathway regulated granular cell proliferation to promote follicle development in Wanxi white goose

Granular cells proliferation in goose regulated by bone morphogenetic proteins (BMPs) signaling pathway is still unknown. In this experiment, BMPs and their receptor, and receptor activated mothers against decapentaplegic homologs (SMADs) were quantitatively expressed in granular cell layer of pre-hierarchycal and hierarchycal follicles in Wanxi White goose. The screened BMP was then used for construction of overexpressed and knockdown vectors and transfected into granular cells of goose to assess the cell proliferation and apoptosis. Granular cells with BMP-overexpressed were then used for ChIP-Seq analysis to elucidate the molecular mechanism of BMP affecting granular cell proliferation. The results showed that the mRNA expression of BMP4 was significantly expressed in pre-hierarchical follicles, and also highly expressed in hierarchical follicles than other BMPs, while the Ⅰ and Ⅱ type of BMP receptors were expressed in basic level. The mRNA expression of SMAD8 was significantly elevated in pre-hierarchical follicles. Overexpression of BMP4 could promote the proliferation of granular cells and inhibited the expression of BMP4 caused a higher cell apoptosis. ChIP-Seq identified multiple regulatory targets of SMAD4, which were mostly related to cell cycle and lipid metabolism according to the GO and KEGG pathway enrichment. From the five most significant binding motif and quantitative expression verification, the activin membrane binding inhibitor (BAMBI) was down regulated in BMP4 overexpressed granular cells. In conclusion, the BMP4 was highly expressed in granular cells and phosphorylates SMAD8, the activated SMAD8 combined with SMAD4 transfers into nucleus to regulate the expression of BAMBI to promote lipid synthesis.

CONGENITAL GRANULAR CELL EPULIS IN ANTERIOR REGION OF THE MANDIBLE: A RARE CASE

Congenital granular cell epulis (CGCE) is a rare benign tumor of unknown etiology, with a predilection for the maxilla. It is characterized by the proliferation of granular cells, exhibiting negative immunohistochemical expression for S-100 protein. An 8-day-old dark-skinned female newborn presented with a nodular lesion located in the anterior region of the mandible. It measured 1.3 × 1.0 × 0.7 cm and had a pedunculated base. It was covered by eroded mucosa and fibrous on palpation. After surgical excision, a diagnosis of congenital granular cell epulis was rendered, based on anatomopathologic and immunohistochemical findings (positive for vimentin and specific neuron enolase and negative for S-100, smooth muscle actin, and desmin). After 18 months of follow-up, there are no signs of recurrence. The recognition of this lesion is important because of functional limitations, such as difficulty in breastfeeding.

WT1 influences apoptosis and proliferation of immature mice granular cells through regulation of the wnt/β-catenin signal pathway

It was to study the influence of Wilms tumor suppressor gene (WT1) on ovarian granular cells (GCs) in mice, and the molecular mechanism involved. LV-WT1 short hairpin ribonucleic acid (shRNA) vector was used to downregulate WT1 expression in granular cells (GCs). The effects of WTI on proliferation and apoptosis of GCs were investigated. Western blot and qRT-PCR were used to assay the mRNA and protein expressions of Bax/bcl-2 in GCs transfected with LV-WT1-RNAi. The expression levels of SUZ12, Wnt5a, Wnt11, Wnt4, Wnt3a, Wnt2 mRNA in GCs were also determined. LV-WT1-RNAi significantly reduced WT1 expression, increased apoptosis and inhibited proliferation of GCs. The inhibition of WT1 had no significant effect on the expression of bcl-2 in GCs. The expressions of Wnt2, Wnt4 and Wnt5a were augmented in WT1-knockdown GCs, relative to non-transfected cells. WT1 activation is necessary for maintaining early survival of GCs in follicles via activation of the Wnt/β-catenin signal pathway.

The effect of follicular fluid from patients with endometriosis, follicle stimulating hormone and bone morphogenetic protein 15 on the proliferation and progesterone secretion of granular cells

Objective: The study was designed to evaluate the effect of follicular fluid from patients with endometriosis, follicle stimulating hormone (FSH), bone morphogenetic protein 15 (BMP-15) on the proliferation and progesterone secretion of human luteinized granular cells in vitro. Methods: Cumulus granulosa cells were collected from the patients who underwent in vitro fertilization and embryo transfer (IVF-ET) ovulation due to tubal or male factor infertility on the day of the retrieval. The cells in the control group were cultured with complete medium of DMEM/F-12, an extra of FSH at a dose of 12 μg/L was added in the FSH group, an extra of BMP-15 at a dose of 6 μg/L was added to the BMP-15 group, an extra of 5% of the follicular fluid from the patients with tubal or male factor infertility was added to the tubal group, an extra of 5% of the follicular fluid from the patients with endometriosis infertility was added to the endometriosis group, an extra of 5% of the follicular fluid from the patients with endometriosis infertility plus FSH at a dose of 12 μg/L were added to the endometriosis plus FSH group, and an extra of 5% of the follicular fluid from the patients with endometriosis infertility plus BMP-15 at a dose of 6 μg/L were added to the endometriosis plus BMP-15 group. Hemacytometer counting method was used to observe the growth of cells after 48 hours, and chemiluminescence method was utilized to measure the level of progesterone in culture supernatant. Results: The cell proliferation was enhanced in the FSH group, while the proliferation was inhibited in the endometriosis group and the endometriosis plus BMP-15 group, compared to the control group, both of which, were statistically significant. Compared to the control group, the progesterone levels from the culture supernatant of granular cells were significantly elevated in the FSH group, tubal group and endometriosis group. The secretion of progesterone in the endometriosis group was lower than that in the tubal group. After addition of FSH into the endometriosis group (the endometriosis plus FSH group), the secretion level of progesterone was significantly increased, compared to the control group and the endometriosis group. After adding BMP-15 into the endometriosis group (the endometriosis plus BMP-15 group), the secretion level of progesterone was increased, compared to the control group. Conclusions: FSH, but not BMP-15, was able to enhance the proliferation and progesterone secretion of granular cells. The proliferation of granular cells and secretion of progesterone were inhibited by the follicular fluid from patients with endometriosis, which was reversed by FSH. However, BMP-15 had no effect on the outcome of follicular fluid from patients with endometriosis.

Protective Effects of Curcumin against Sodium Arsenite-induced Ovarian Oxidative Injury in a Mouse Model.

Background:Excessive reactive oxygen species (ROS) may lead to a number of reproductive diseases such as polycystic ovary syndrome. This study aimed to establish an animal model of ovarian oxidative stress and to assess the protective effect of curcumin against oxidative injury.Methods:Ovarian oxidative stress was induced in female Kunming mice (n = 40) with intraperitoneal injection of 8 mg/kg sodium arsenite (As) once every other day for 16 days; meanwhile, they were, respectively, treated by intragastric administration of 0, 100, 150, or 200 mg/kg (n = 10/group) curcumin once per day for 21 days. Ten normal mice were used as control. Then, the mice were injected intraperitoneally with BrdU and sacrificed; the right ovaries were collected for hematoxylin and eosin (HE) staining and BrdU immunohistochemistry, and the left ovaries for enzyme-linked immunosorbent assay (ELISA) and Western blotting analyses.Results:The ELISA results showed that ROS (11.74 ± 0.65 IU/mg in 8 mg/kg AS + 0 mg/kg curcumin group vs. 10.71 ± 0.91 IU/mg in control group, P = 0.021) and malondialdehyde (MDA) (0.32 ± 0.02 nmol/g in 8 mg/kg AS + 0 mg/kg curcumin group vs. 0.27 ± 0.02 nmol/g in control group, P = 0.048) increased while superoxide dismutase (SOD) (3.96 ± 0.36 U/mg in 8 mg/kg AS + 0 mg/kg curcumin group vs. 4.51 ± 0.70 U/mg in control group, P = 0.012) and glutathione peroxidase (17.36 ± 1.63 U/g in 8 mg/kg AS + 0 mg/kg curcumin group vs. 18.92 ± 1.80 U/g in control group, P = 0.045) decreased in the ovary after injection of As, indicating successful modeling of oxidative stress. Curcumin treatment could considerably increase SOD (4.57 ± 0.68, 4.49 ± 0.27, and 4.56 ± 0.25 U/mg in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively, all P < 0.05) while significantly reduce ROS (10.64 ± 1.38, 10.73 ± 0.71, and 10.67 ± 1.38 IU/mg in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively, all P < 0.05) and MDA (0.28 ± 0.02, 0.25 ± 0.03, and 0.27 ± 0.04 nmol/g in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively; both P < 0.05) in the ovary. HE staining and BrdU immunohistochemistry of the ovarian tissues indicated the increased amount of atretic follicles (5.67 ± 0.81, 5.84 ± 0.98, and 5.72 ± 0.84 in 100 mg/kg, 150 mg/kg, and 200 mg/kg curcumin group, respectively, all P < 0.05), and the inhibited proliferation of granular cells under oxidative stress would be reversed by curcumin. Furthermore, the Western blotting of ovarian tissues showed that the p66Shc expression upregulated under oxidative stress would be lowered by curcumin.Conclusion:Curcumin could alleviate arsenic-induced ovarian oxidative injury to a certain extent.

口腔領域におけるか粒細胞腫の臨床病理組織学的検討 その組織発生についての考察

Granular cell tumors (GCTs) are rare, soft tissue tumors, and their histogenesis is still controversial, although the neurogenic theory, especially Schwann cell derivation, is considered the most plausible at present.We studied eight cases of GCT clinicopathologically and immunohistochemically. The results were as follows:1. Granular cells of GCT had a variety of sizes and shapes, were filled with fine eosinophilic granules, and showed lysosome-like characteristics, also with a variety of sizes and shapes, on electron microscopic observation.2. Immunohistochemically, the intracytoplasmic granules showed positive reactions for S-100 protein, NSE, Kp-1 and diastase-resistant PAS staining but a negative reaction for a 1-antichymotrypsin. Additional lipid staining, such as oil red O, Sudan black B, and Nile blue, was performed in two cases, but was all negative.3. In three cases, an appearance of transition from striated muscle fibers to granular cells, and the accumulation of intracytoplasmic granules with the shape of striated muscle fibers remaining were seen at the margin of GCT masses. In addition, four cases out of eight showed weakly positive reactions for myoglobin.Thus, several cases were suggested to have a myogenic origin, and neither 5-100 protein nor NSE provided conclusive evidence of neurogenic origin due to their poor specificity. It was, therefore difficult to limit their derivation to neurogenic origin, and further examinations are required.The accumulation of intracytoplasmic granules in other tumors, such as neurofibroma, which was referred to as granular cell neurofibroma by Enzinger and Weiss (1983), and leiomyoma, were previously reported. The neoplastic proliferation of granular cells seems unlikely in view of their few cell organelles on electron microscopic observation.In order to explain our results and those of previous studies, it is most reasonable to consider that GCT is not a neoplastic lesion, that the accumulation of intracytoplasmic granules in GCT is the result of degenerative or reprocessing changes (Whitten, 1968), and that such “granular cell changes” occur in various tissues (Shear, 1960 and Millar, 1977).Furthermore, previously reported “malignant” GCTs, which showed histological malignancy and remote metastasis and were often diagnosed with much difficulty, may include pre-existing malignant tumors with “granular cell changes”.