scholarly journals Role of Gonadotropin-Releasing Hormone as an Autocrine Growth Factor in Human Ovarian Surface Epithelium1

Endocrinology ◽  
2000 ◽  
Vol 141 (1) ◽  
pp. 72-80 ◽  
Author(s):  
Sung Keun Kang ◽  
Kyung-Chul Choi ◽  
Kwai Wa Cheng ◽  
Parimal S. Nathwani ◽  
Nelly Auersperg ◽  
...  
Keyword(s):  
Gnrh Agonist ◽  
Messenger Rna ◽  
Ovarian Surface Epithelium ◽  
Inhibitory Effect ◽  
Surface Epithelium ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Rt Pcr ◽  
Ovarian Surface ◽  
Ovarian Surface Epithelial

Abstract Epithelial ovarian cancer, which accounts for 80–90% of all ovarian cancers, is the most common cause of death from gynecological malignancies and is believed to originate from the ovarian surface epithelium. In the present study we investigated the expression of GnRH and its receptor in human ovarian surface epithelial (hOSE) cells and provided novel evidence that GnRH may have antiproliferative effects in this tissue. Using RT-PCR and Southern blot analysis, we cloned the GnRH and GnRH receptor (GnRHR) in hOSE cells. Sequence analysis revealed that GnRH and its receptor have sequences identical to those found in the hypothalamus and pituitary, respectively. To address whether GnRH regulates its own and receptor messenger RNA (mRNA), the cells were treated with different concentrations of the GnRH agonist (d-Ala6)-GnRH. Expression levels of GnRH and its receptor were investigated using quantitative and competitive RT-PCR, respectively. Interestingly, a biphasic effect was observed for the GnRH and GnRHR mRNA levels. High concentrations of the GnRH agonist (10−7 and 10−9m) decreased GnRH and GnRHR mRNA levels, whereas a low concentration (10−11m) resulted in up-regulation of GnRH and receptor mRNA levels. Treatment with the GnRH antagonist, antide, prevented the biphasic effects of the GnRH agonist in hOSE cells, confirming the specificity of the response. Furthermore, to investigate the physiological significance, we studied receptor-mediated growth regulatory effects of GnRH in human ovarian surface epithelial cells. The cells were treated with GnRH analogs, and the proliferative index of cells was measured using a [3H]thymidine incorporation assay. (d-Ala6)-GnRH had a direct inhibitory effect on the growth of hOSE cells in a time- and dose-dependent manner. This antiproliferative effect of the GnRH agonist was receptor mediated, as cotreatment of hOSE cells with antide abolished the growth inhibitory effects of the GnRH agonist. The results strongly suggest that GnRH can act as an autocrine/paracrine regulator in hOSE cells.

333 GENERATION OF OOCYTE-LIKE STRUCTURE FROM OVARIAN SURFACE EPITHELIAL STEM CELLS OF GOAT

2015 ◽  
Vol 27 (1) ◽  
pp. 255 ◽  
Author(s):  
S. Fatima ◽  
V. Sharma ◽  
S. Saini ◽  
S. Saugandhika ◽  
H. N. Malik ◽  
...  
Keyword(s):  
Stem Cells ◽  
Endangered Species ◽  
Epithelial Cells ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Epithelial Stem Cells ◽  
Rt Pcr ◽  
Ovarian Surface ◽  
Ovarian Surface Epithelial

Stem cells have potential for therapeutic application. Continuous repair of ovarian surface epithelium following folliculogenesis and ovarian carcinoma suggests the presence of stem cells in ovarian epithelial cells. In vitro gametogenesis in livestock will result in large numbers of oocytes production from a single ovary, resulting in faster multiplication of superior germplasm of livestock species, treatment of infertile animals, and conservation of endangered species. The present study was conducted with the objective of in vitro differentiation of putative ovarian surface epithelial stem cells into oocyte-like structures in goat model. Ovary samples of 1- to 2-year-old goats were collected from slaughterhouse. The surface of the ovary was gently scraped using sterile blunt scraper to isolate ovarian surface epithelial stem cells. These scraped cells were cultured in DMEM/F12 supplemented with 20% FBS for 3 weeks in 5% CO2 at 37°C with maximum humidity. The cultured stem cells were characterised for stemness by RT-PCR and immunostaining for Oct4, Sox2, and Nanog genes after 3 weeks. These putative stem cells were in vitro differentiated spontaneously to oocyte-like structures in DMEM/F12 medium and characterised for premeiotic markers by RT-PCR and immunostaining for VASA, DAZL, and STELLA genes. Results of this study provide evidence for the presence of putative stem cells with pluripotent characteristics in the ovarian surface epithelium. The cultured cells were found to be round in shape, with a high nucleus to cytoplasm ratio under inverted microscope, and found positive for stem cell markers of Oct4, Sox2, and Nanog genes. A total of 66 oocyte-like structures were produced from 12 ovaries. These oocyte-like structures were nearly similar to oocytes produced in vivo, both morphologically and in molecular gene expression. The oocyte-like structures were also found positive for premeiotic markers of VASA, DAZL, and STELLA genes by RT-PCR and immunostaining. From this study, we concluded that the ovarian surface epithelial cells have putative stem cells which can be in vitro differentiated into oocyte-like structures in goat. These oocyte-like structures need further characterisation of their surface membrane, more molecular markers, and following their developmental potential. These oocytes can help for multiplication of elite germplasm, curing infertile animals, and saving endangered species.

Isolation and Characterization of Mesenchymal Stem Cells from Canine Ovarian Surface Epithelium

2021 ◽  
Author(s):  
Ajeet Kumar Jha ◽  
Anirban Mandal ◽  
Kalyani Ray ◽  
Shyamal Kanti Guha
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Stromal Vascular Fraction ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
High Plasticity ◽  
Rt Pcr ◽  
Differentiation Potential ◽  
Ovarian Surface ◽  
Isolation And Characterization

Background: Few studies have confirmed the presence of ovarian tissue stem cells indicating the capacity for differentiation. Based on this fact, it was hypothesized that mesenchymal stem cells (MSC) were found in ovarian surface epithelium (OSE) of canines that could easily be isolated. Methods: Both left and right ovaries were minced and digested using collagenase to obtain a stromal vascular fraction (SVF). MSCs were characterized using RT-PCR. To ascertain the trilineage differentiation potential, MSCs were stained with respective stain for osteocytes, chondrocytes and adipocytes. Result: We observed elongated, spindle-shaped and fibroblast like appearance of cells after 72 h of initial culture. Expression of MSC specific surface markers were observed through RT-PCR. Using Stem Pro® differentiation medium, OSE were differentiated into osteogenic, chondrogenic and adipogenic lineages and were found to be potential source for isolation, characterization and differentiation of MSCs. Canine (OSE) is easily accessible, multipotent and has high plasticity, holding promise for applications in regenerative medicine.

scholarly journals Loss of GATA6 Leads to Nuclear Deformation and Aneuploidy in Ovarian Cancer

2009 ◽  
Vol 29 (17) ◽  
pp. 4766-4777 ◽  
Author(s):  
Callinice D. Capo-chichi ◽  
Kathy Q. Cai ◽  
Joseph R. Testa ◽  
Andrew K. Godwin ◽  
Xiang-Xi Xu
Keyword(s):  
Ovarian Cancer ◽  
Nuclear Envelope ◽  
Cancer Cells ◽  
Human Cancer ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Ovarian Cancer Cells ◽  
Ovarian Surface ◽  
Ovarian Surface Epithelial ◽  
Ovarian Tumorigenesis

ABSTRACT A prominent hallmark of most human cancer is aneuploidy, which is a result of the chromosomal instability of cancer cells and is thought to contribute to the initiation and progression of most carcinomas. The developmentally regulated GATA6 transcription factor is commonly lost in ovarian cancer, and the loss of its expression is closely associated with neoplastic transformation of the ovarian surface epithelium. In the present study, we found that reduction of GATA6 expression with small interfering RNA (siRNA) in human ovarian surface epithelial cells resulted in deformation of the nuclear envelope, failure of cytokinesis, and formation of polyploid and aneuploid cells. We further discovered that loss of the nuclear envelope protein emerin may mediate the consequences of GATA6 suppression. The nuclear phenotypes were reproduced by direct suppression of emerin with siRNA. Thus, we conclude that diminished expression of GATA6 leads to a compromised nuclear envelope that is causal for polyploidy and aneuploidy in ovarian tumorigenesis. The loss of emerin may be the basis of nuclear morphological deformation and subsequently the cause of aneuploidy in ovarian cancer cells.

scholarly journals Regulation of Aromatase Expression in Human Ovarian Surface Epithelial Cells1

2000 ◽  
Vol 85 (12) ◽  
pp. 4889-4899
Author(s):  
Tomoharu Okubo ◽  
Samuel C. Mok ◽  
Shiuan Chen
Keyword(s):  
Protein Kinase ◽  
Epithelial Cells ◽  
Messenger Rna ◽  
Phorbol Esters ◽  
Pcr Analysis ◽  
Aromatase Activity ◽  
Rt Pcr ◽  
T7 Promoter ◽  
Ovarian Surface ◽  
Ovarian Surface Epithelial

Ovarian cancer originates mainly from surface epithelial cells, which are potential targets of estrogen action. Using immunohistochemistry and RT-PCR analysis, aromatase (estrogen synthetase) can be detected in human ovarian surface epithelial tumors. In this study, we functionally characterized the aromatase expressed in a primary cell culture, normal human ovarian surface epithelial (HOSE) 17. The apparent Km and Vmax values were determined to be 5.8 ± 0.5 nm, and 0.3 ± 0.0 pmol/mg·h, respectively. The aromatase activity in HOSE 17 cells can be induced effectively by phorbol esters and forskolin, suggesting that estrogen biosynthesis in HOSE 17 cells is mainly regulated through protein kinase C- and protein kinase A-mediated mechanisms. Exon I-specific RT-PCR revealed that phorbol esters predominantly up-regulated promoter II. Whereas forskolin treatment increased exon I.3A-containing messenger RNA, the aromatase activity remained low in the cells treated with this agent. In vitro transcription/translation analysis using plasmids containing T7 promoter and the human snail gene (SnaH) as a reporter capped with different untranslated exon Is revealed that exon PII-containing transcripts were translated more effectively than exon I.3-containing transcripts. These findings explain why aromatase activity is higher in cells with the PII-containing transcripts than is cells with the I.3-containing transcripts. Our results indicate that aromatase is functionally expressed in human ovarian surface epithelial cells and its expression is regulated at both the transcriptional and translational levels.

scholarly journals Differential regulation of two forms of gonadotropin-releasing hormone messenger ribonucleic acid by gonadotropins in human immortalized ovarian surface epithelium and ovarian cancer cells

2006 ◽  
Vol 13 (2) ◽  
pp. 641-651 ◽  
Author(s):  
Jung-Hye Choi ◽  
Kyung-Chul Choi ◽  
Nelly Auersperg ◽  
Peter C K Leung
Keyword(s):  
Ovarian Cancer ◽  
Cell Lines ◽  
Ovarian Cancer Cell ◽  
Ovarian Surface Epithelium ◽  
Gonadotropin Releasing Hormone ◽  
Surface Epithelium ◽  
Ovarian Cancer Cells ◽  
Mrna Levels ◽  
Ovarian Surface ◽  
Ovarian Cancer Cell Lines

Although gonadotropin-releasing hormone (GnRH) has been shown to play a role as an autocrine/ paracrine regulator of cell growth in ovarian surface epithelium and ovarian cancer, the factors which regulate the expression of GnRH and its receptor in these cells are not well characterized. In the present study, we employed real-time PCR to determine the potential regulatory effect of gonadotropins on the expression levels of GnRH I (the mammalian GnRH), GnRH II (a second form of GnRH) and their common receptor (GnRHR) in immortalized ovarian surface epithelial (IOSE-80 and IOSE-80PC) cells and ovarian cancer cell lines (A2780, BG-1, CaOV-3, OVCAR-3 and SKOV-3). The cells were treated with increasing concentrations (100 and 1000 ng/ml) of recombinant follicle-stimulating hormone (FSH) or luteinizing hormone (LH) for 24 h. Treatment with FSH or LH reduced GnRH II mRNA levels in both IOSE cell lines and in three out of five ovarian cancer cell lines (A2780, BG-1 and OVCAR-3). A significant decrease in GnRHR mRNA levels was observed in IOSE and ovarian cancer cells, except CaOV-3 cells, following treatment with FSH or LH. In contrast, treatment with either FSH or LH had no effect on GnRH I mRNA levels in these cells, suggesting that gonadotropins regulate the two forms of GnRH and its receptor differentially. In separate experiments, the effect of gonadotropins on the anti-proliferative action of GnRH I and GnRH II agonists in IOSE-80, OVCAR-3 and SKOV-3 cells was investigated. The cells were pretreated with FSH or LH (100 ng/ml) for 24 h after which they were treated with either GnRH I or GnRH II (100 ng/ml) for 2 days, and cell growth was assessed by the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide] assay. Pretreatment of the cells with FSH or LH significantly reversed the growth inhibitory effect of GnRH I and GnRH II agonists in these cell types. These results provide the first demonstration of a potential interaction between gonadotropins and the GnRH system in the growth regulation of normal ovarian surface epithelium and its neoplastic counterparts.

A BRIEF COMMUNICATION

2005 ◽  
Vol 230 (6) ◽  
pp. 429-433 ◽  
Author(s):  
William J. Murdoch ◽  
Edward A. Van Kirk ◽  
Brenda M. Alexander
Keyword(s):  
Ovarian Cancer ◽  
Epithelial Cells ◽  
Laying Hens ◽  
Ovarian Surface Epithelium ◽  
Egg Laying ◽  
High Rate ◽  
Surface Epithelium ◽  
Ovarian Surface ◽  
Guanine Adducts ◽  
Ovarian Surface Epithelial

A cause-effect relationship between ovulation and common (surface) epithelial ovarian cancer has been suspected for many years. The ovarian surface epithelium apparently becomes exposed to genotoxins that are generated during the ovulatory process. Intensive egg-laying hens readily develop ovarian carcinomatosis. Indeed, elevated levels of potentially mutagenic 8-oxo-guanine adducts were detected in avian ovarian epithelial cells isolated from the apical surfaces and perimeters of pre-and postovulatory follicles, respectively. Internucleosomal DNA fragmentation indicative of apoptosis was evident in ovarian surface epithelial cells associated with the formative site of ovulation (stigma line) and regressive ruptured follicles. It is conceivable that a genetically altered progenitor cell with unrepaired DNA but not committed to death (i.e., a unifocal “escape”) could give rise to a transformed phenotype. Hence, the high rate of ovarian cancer in egg-laying hens could be the consequence of genomic damages to the ovarian surface epithelium associated with incessant ovulations, thereby increasing the likelihood of mutation and clonal expansion.

scholarly journals Estrogen Receptor Alpha (ER-α) and Beta (ER-β) mRNAs in Normal Ovary, Ovarian Serous Cystadenocarcinoma and Ovarian Cancer Cell Lines: Down-Regulation of ER-β in Neoplastic Tissues

1998 ◽  
Vol 83 (3) ◽  
pp. 1025-1028 ◽  
Author(s):  
Alfred W. Brandenberger ◽  
Meng Kian Tee ◽  
Roberts B. Jaffe
Keyword(s):  
Ovarian Cancer ◽  
Estrogen Receptor ◽  
Granulosa Cells ◽  
Ovarian Cancer Cell ◽  
Ovarian Surface Epithelium ◽  
Surface Epithelium ◽  
Mrna Levels ◽  
Rt Pcr ◽  
Low Level ◽  
Ovarian Cancer Cell Lines

The prognosis in ovarian carcinoma, the most lethal of the gynecologic neoplasms, is poor and has changed little in the last three decades. Only a small number respond to antiestrogen therapy, although the classic estrogen receptor, ER-α, has been identified in ovarian surface epithelium, from which approximately 90% of ovarian cancers originate. We have previously shown that ER-β mRNA is most abundant in human fetal ovaries, suggesting that it might play an important role in ovarian development. Therefore, we investigated the mRNA levels of both ERs in normal ovaries, ovarian serous cystadenocarcinomas, granulosa cells from patients undergoing in vitro fertilization (IVF), the ovarian surface epithelium cell line IOSE-Van, and the ovarian cancer cell lines SKOV3, HEY and OCC1. Northern blots of normal and neoplastic ovaries were hybridized with an ER-β riboprobe that spans the A/B domain. We detected two major hybridizing bands at approximately 8 and 10 kb. An RNase protection assay using the same probe revealed a single band of the expected size. Hybridizing the same blot with an ER-α riboprobe showed a strong hybridizing band at approximately 6.5 kb. In ovarian cancer samples, ER-β mRNA level was decreased when compared to normal ovaries. Using 25 cycles of RT-PCR followed by Southern blotting, we found equal amounts of ER-α and -β mRNAs in normal ovaries in all age groups from 33 to 75 years; however, in ovarian cancer tissue, the level of ER-α mRNA was similar or slightly higher, comparable to 103 to 104 copies of plasmid DNA, but ER-β mRNA levels were markedly decreased. Granulosa cells from IVF patients expressed high levels of ER-β mRNA. The OSE cell line expressed low level of ER-α, detectable after 40 cycles of RT-PCR and no ER-β mRNA. SKOV3, showed low level of ER-α and β mRNAs, whereas OCC1 showed low level of ER-β and relatively high level of ER-α. HEY did not contain detectable amounts of either ER after 40 cycles of RT-PCR. We found no evidence of differential splicing or major deletions in almost the entire coding region of ER-β in either normal ovaries or tumor samples.

scholarly journals Antiinflammatory Steroid Action in Human Ovarian Surface Epithelial Cells

2004 ◽  
Vol 89 (9) ◽  
pp. 4538-4544 ◽  
Author(s):  
Michael T. Rae ◽  
Deborah Niven ◽  
Hilary O. D. Critchley ◽  
Christopher R. Harlow ◽  
Stephen G. Hillier
Keyword(s):  
Glucocorticoid Receptor ◽  
Mrna Expression ◽  
Mrna Level ◽  
Inhibitory Effect ◽  
Surface Epithelium ◽  
Pcr Analysis ◽  
Mrna Levels ◽  
Cell Monolayers ◽  
Ovarian Surface ◽  
Cox 2

The human ovarian surface epithelium (OSE) is subject to serial injury and repair during ovulation, which is a natural inflammatory event. We asked whether there is a compensatory antiinflammatory component to this process, involving steroid hormones produced locally at the time of ovulation. Quantitative RT-PCR analysis of total RNA from cultured human OSE cell monolayers showed that exposure to proinflammatory IL1α (500 pg/ml) increased mRNA levels of cyclooxygenase-2 (COX-2) (P < 0.01) at 48 h. The COX-2 mRNA response to IL1α was associated with an approximate 18-fold (P < 0.01) increase in mRNA levels of 11β-hydroxysteroid dehydrogenase type 1 (11βHSD1), encoding the steroid dehydrogenase that reversibly reduces cortisone to antiinflammatory cortisol. Addition of cortisol to OSE cell culture medium dose-dependently suppressed the COX-2 mRNA response to IL1α (P < 0.01) but reciprocally enhanced the 11βHSD1 mRNA response (P < 0.05), with both effects strongest at 1 μm cortisol. Presence of glucocorticoid receptor-α mRNA and protein was established in OSE cell monolayers and treatment with IL1α shown to significantly up-regulate the glucocorticoid receptor-α mRNA level (P < 0.05). Glucocorticoid receptor antagonist (RU486, 10 μm) fully reversed the inhibitory effect of 1 μm cortisol on IL1α-stimulated COX-2 mRNA expression. Progesterone also suppressed IL1α-induced COX-2 mRNA expression but had no significant effect on IL1α-stimulated 11βHSD1 expression. These data provide direct evidence for antiinflammatory actions of cortisol and progesterone in human OSE cells.

scholarly journals Discovery of 3-Arylquinoxaline Derivatives as Potential Anti-Dengue Virus Agents

2019 ◽  
Vol 20 (19) ◽  
pp. 4786
Author(s):  
Chih-Hua Tseng ◽  
Cheng-Ruei Han ◽  
Kai-Wei Tang
Keyword(s):  
Dengue Virus ◽  
Messenger Rna ◽  
Active Sites ◽  
Hydrophobic Interactions ◽  
Biological Activities ◽  
Inhibitory Effect ◽  
Docking Studies ◽  
Mrna Levels ◽  
Dependent Manner ◽  
Cox 2

We designed and synthesized a series of novel 3-arylquinoxaline derivatives and evaluated their biological activities as potential dengue virus (DENV) replication inhibitors. Among them, [3-(4-methoxyphenyl)quinoxalin-2-yl](phenyl)methanol (19a), [6,7-dichloro-3-(4-methoxyphenyl)quinoxalin-2-yl](phenyl)methanol (20a), and (4-methoxyphenyl)(3-phenylquinoxalin-2-yl)methanone (21b) were found to significantly inhibit the DENV RNA expression in Huh-7-DV-Fluc cells with a potency better than that of ribavirin. Compound 19a reduced DENV replication in both viral protein and messenger RNA (mRNA) levels in a dose-dependent manner and exhibited no significant cell cytotoxicity. Notably, compound 19a exhibited a half maximal effective concentration (EC50) value at 1.29 ± 0.74 μM. We further observed that the inhibitory effect of 19a on DENV replication was due to suppression of DENV-induced cyclooxygenase-2 (COX-2) expression. Docking studies also showed that 19a caused hydrophobic interactions at the active sites with Arg29, Glu31, Tyr116, Leu138, Pro139, Lys454, Arg455, and Gln529. The calculated lowest binding energy between the 19a and COX-2 was −9.10 kcal/mol. In conclusion, compound 19a might be a potential lead compound for developing an anti-DENV agent.

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