17β-Estradiol overcomes human myeloma RPMI8226 cell suppression of growth, ALP activity, and mineralization in rat osteoblasts and improves RANKL/OPG balance in vitro

It is well known that diabetes affects bone in human and animal models, and leads to osteopenia and osteoporosis. Bone-mineral density and other biochemical markers of bone turnover are very much affected in people with diabetes. Reduced bone mass, occurring with increased frequency in diabetes mellitus, has been attributed to poor glycemic control, but the pathogenic mechanisms remain unknown. High concentrations of glucose (hyperglycemia) in diabetics leads to this complication. Very few in vitro studies using bone-cell lines have been carried out to address this problem. In this study, we examined the effects of different doses of glucose concentration (5.5, 16.5, and 49.4 mmol/L), alone, with insulin (0.6 µg/mL), or with 17β-estradiol (E2) (10 nmol/L), on rat bone-marrow stromal cells (BMSCs) in the presence of an osteogenic medium. BMSC proliferation and alkaline phosphatase (ALP) were studied after 3 and 7 d of culture, respectively; the area stained for collagen and mineralized nodules was studied after 28 d of culture. With high concentrations of glucose, BMSC proliferation, ALP activity, the number of nodules formed, and the area stained for collagen were greatly reduced. Insulin treatment alone was able to increase [3H]-thymidine uptake or ALP activity, whereas both insulin and estradiol were able to increase the number of mineralized nodules and the area stained for collagen and mineralization. In conclusion, this study suggests that insulin and estradiol are able to contain the deleterious effect of high concentrations of glucose on BMSC-derived osteoblast proliferation and function.Key words: bone marrow cells, estradiol, glucose, insulin, mineralization.

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Electrochemotherapy with Calcium Chloride and 17β-Estradiol Modulated Viability and Apoptosis Pathway in Human Ovarian Cancer

Pharmaceutics ◽

10.3390/pharmaceutics13010019 ◽

2020 ◽

Vol 13 (1) ◽

pp. 19

Author(s):

Zofia Łapińska ◽

Michał Dębiński ◽

Anna Szewczyk ◽

Anna Choromańska ◽

Julita Kulbacka ◽

...

Keyword(s):

Ovarian Cancer ◽

Cancer Cells ◽

Calcium Chloride ◽

Morphological Changes ◽

Cell Membrane Permeability ◽

Immunocytochemical Staining ◽

Ovarian Cancer Cells ◽

Apoptosis Pathway ◽

17Β Estradiol

Estrogens (Es) play a significant role in the carcinogenesis and progression of ovarian malignancies. Depending on the concentration, Es may have a protective or toxic effect on cells. Moreover, they can directly or indirectly affect the activity of membrane ion channels. In the presented study, we investigated in vitro the effectiveness of the ovarian cancer cells (MDAH-2774) pre-incubation with 17β-estradiol (E2; 10 µM) in the conventional chemotherapy (CT) and electrochemotherapy (ECT) with cisplatin or calcium chloride. We used three different protocols of electroporation including microseconds (µsEP) and nanoseconds (nsEP) range. The cytotoxic effect of the applied treatment was examined by the MTT assay. We used fluorescent staining and holotomographic imaging to observe morphological changes. The immunocytochemical staining evaluated the expression of the caspase-12. The electroporation process’s effectiveness was analyzed by a flow cytometer using the Yo-Pro™-1 dye absorption assay. We found that pre-incubation of ovarian cancer cells with 17β-estradiol may effectively enhance the chemo- and electrochemotherapy with cisplatin and calcium chloride. At the same time, estradiol reduced the effectiveness of electroporation, which may indicate that the mechanism of increasing the effectiveness of ECT by E2 is not related to the change of cell membrane permeability.

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17β-Estradiol Reduces Cardiomyocyte Apoptosis In Vivo and In Vitro via Activation of Phospho-Inositide-3 Kinase/Akt Signaling

Circulation Research ◽

10.1161/01.res.0000144126.57786.89 ◽

2004 ◽

Vol 95 (7) ◽

pp. 692-699 ◽

Cited By ~ 223

Author(s):

Richard D. Patten ◽

Isaac Pourati ◽

Mark J. Aronovitz ◽

Jason Baur ◽

Flore Celestin ◽

...

Keyword(s):

Akt Signaling ◽

Cardiomyocyte Apoptosis ◽

17Β Estradiol

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Role of leukemia inhibitor factor (LIF) in decidualisation of murine uterine stromal cells in vitro

Journal of Endocrinology ◽

10.1677/joe.0.1810477 ◽

2004 ◽

Vol 181 (3) ◽

pp. 477-492 ◽

Cited By ~ 19

Author(s):

AA Fouladi Nashta ◽

CV Andreu ◽

N Nijjar ◽

JK Heath ◽

SJ Kimber

Keyword(s):

In Vitro Culture ◽

Stromal Cells ◽

Control Level ◽

Calf Serum ◽

Prostaglandin E ◽

Dependent Manner ◽

Alp Activity ◽

Dose Dependent ◽

In Cells

Decidualisation of uterine stromal cells is a prerequisite for implantation of the embryo in mice. Here we have used an in vitro culture system in which stromal cells decidualise as indicated by a number of markers, including an increase in alkaline phosphatase (ALP) activity. The latter was used as a quantitative marker of decidualisation in the presence of low (2%) fetal calf serum. Prostaglandin E(2) (PGE(2)), which is known to induce decidualisation, increased ALP activity, and this effect was blocked in a dose-dependent manner by indomethacin. Leukemia inhibitory factor (LIF) was then examined, but it had no effect on PGE(2) secretion. However, LIF suppressed ALP activity in a dose-dependent manner in the presence of 2% serum, while an inhibitor of LIF that competes for binding to its receptor reversed the effect of LIF and increased ALP activity above the control level. In serum-free cultures, stromal cells differentiated rapidly, and no differences were observed between LIF-treated and untreated cultures. Stromal cells produce LIF during in vitro culture, and this peaked at 48 h. Freshly collected stromal cells from both day-2 and -4 pregnant mice expressed mRNA for the LIF receptor, and the transcript level was higher in cells isolated on day 4. However, no differences were observed in the relative levels of transcripts in cells from day 2 and day 4 after culture, nor were there differences between the LIF-treated cultures and controls. Therefore, in this study, we have shown that LIF suppresses decidualisation of murine uterine stromal cells in the presence of serum, this is not due to the regulation of PGE(2) secretion by stromal cells.

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Feasibility of Secondary Follicle Isolation, Culture and Achievement of In-Vitro Oocyte Maturation from Superovulated Ovaries: An Experimental Proof-of-Concept Study Using Mice

Journal of Clinical Medicine ◽

10.3390/jcm10132757 ◽

2021 ◽

Vol 10 (13) ◽

pp. 2757

Author(s):

Xia Hao ◽

Amandine Anastácio ◽

Kenny A. Rodriguez-Wallberg

Keyword(s):

Oocyte Maturation ◽

In Vitro Maturation ◽

Follicular Development ◽

Clinical Situation ◽

Experimental Proof ◽

Secondary Follicle ◽

17Β Estradiol ◽

Follicle Size ◽

And Control

Fertility preservation through ovarian stimulation, aiming at cryopreserving mature oocytes or embryos, is sometimes unsuccessful. This clinical situation deserves novel approaches to overcome infertility following cancer treatment in patients facing highly gonadotoxic treatment. In this controlled experimental study, we investigated the feasibility of in-vitro culturing secondary follicles isolated from superovulated ovaries of mice recently treated with gonadotropins. The follicle yields of superovulated ovaries were 45.9% less than in unstimulated controls. Follicles from superovulated ovaries showed faster growth pace during the initial 7 days of culture and secreted more 17β-estradiol by the end of culture vs controls. Parameters reflecting the outcome of follicular development and oocyte maturation competence in vitro were similar between superovulated and control groups, with a similar follicle size at the end of culture and around 70% survival. Nearly half of cultured follicles met the criteria for in-vitro maturation in both groups and approximately 60% of those achieved a mature MII oocyte, similarly in both groups. Over 60% of obtained MII oocytes displayed normal-looking spindle and chromosome configurations, without significant differences between the groups. Using a validated follicle culture system, we demonstrated the feasibility of secondary follicle isolation, in-vitro culture and oocyte maturation with normal spindle and chromosome configurations obtained from superovulated mice ovaries.

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A New MAP Kinase Protein Involved in Estradiol-Stimulated Reproduction of the Helminth ParasiteTaenia crassiceps

Journal of Biomedicine and Biotechnology ◽

10.1155/2010/747121 ◽

2010 ◽

Vol 2010 ◽

pp. 1-11 ◽

Cited By ~ 3

Author(s):

Galileo Escobedo ◽

Gloria Soldevila ◽

Guadalupe Ortega-Pierres ◽

Jesús Ramsés Chávez-Ríos ◽

Karen Nava ◽

...

Keyword(s):

Flow Cytometry ◽

Map Kinases ◽

Host Cells ◽

Cross Contamination ◽

Flow Cytometry Analysis ◽

Cellular Processes ◽

17Β Estradiol ◽

Activation Motif

MAP kinases (MAPK) are involved in the regulation of cellular processes such as reproduction and growth. In parasites, the role of MAPK has been scarcely studied. Here, we describe the participation of an ERK-like protein in estrogen-dependent reproduction of the helminth parasiteTaenia crassiceps. Our results show that 17β-estradiol induces a concentration-dependent increase in the bud number of in vitro cultured cysticerci. If parasites are also incubated in presence of an ERK-inhibitor, the stimulatory effect of estrogen is blocked. The expression of ERK-like mRNA and its corresponding protein was detected in the parasite. The ERK-like protein was over-expressed by all treatments. Nevertheless, a strong induction of phosphorylation of this protein was observed only in response to 17β-estradiol. Cross-contamination by host cells was discarded by flow cytometry analysis. Parasite cells expressing the ERK-like protein were exclusively located at the subtegument tissue by confocal microscopy. Finally, the ERK-like protein was separated by bidimensional electrophoresis and then sequenced, showing the conserved TEY activation motif, typical of all known ERK 1/2 proteins. Our results show that an ERK-like protein is involved in the molecular signalling during the interaction between the host andT. crassiceps, and may be considered as target for anti-helminth drugs design.

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Behavior of fetal rat osteoblasts cultured in vitro on the DP-bioactive glass substratum

Materials Chemistry and Physics ◽

10.1016/s0254-0584(97)80174-5 ◽

1997 ◽

Vol 49 (3) ◽

pp. 270-276 ◽

Cited By ~ 10

Author(s):

Feng-Huei Lin ◽

Chun-Jen Liao ◽

Haw-Chang Liu ◽

Kao-Shau Chen ◽

Jui-Sheng Sun

Keyword(s):

Bioactive Glass ◽

Fetal Rat ◽

Rat Osteoblasts

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Effects of cyclosporin A on rat osteoblasts (ROS 17/2.8 cells) in vitro

Calcified Tissue International ◽

10.1007/bf00334490 ◽

1992 ◽

Vol 51 (4) ◽

pp. 291-297 ◽

Cited By ~ 36

Author(s):

Laurie K. McCauley ◽

Thomas J. Rosol ◽

Charles C. Capen

Keyword(s):

Cyclosporin A ◽

Rat Osteoblasts

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Biocompatibility evaluation in vitro. Part I: Morphology expression and proliferation of human and rat osteoblasts on the biomaterials

Journal of Central South University of Technology ◽

10.1007/s11771-001-0015-6 ◽

2001 ◽

Vol 8 (1) ◽

pp. 1-8 ◽

Cited By ~ 3

Author(s):

Jian-ming Ruan ◽

Grant M. Helen

Keyword(s):

Rat Osteoblasts

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An integrated approach to unravel a crucial structural property required for the function of the insect steroidogenic Halloween protein Noppera-bo

Journal of Biological Chemistry ◽

10.1074/jbc.ra119.011463 ◽

2020 ◽

Vol 295 (20) ◽

pp. 7154-7167 ◽

Cited By ~ 1

Author(s):

Kotaro Koiwai ◽

Kazue Inaba ◽

Kana Morohashi ◽

Sora Enya ◽

Reina Arai ◽

...

Keyword(s):

Structural Characteristics ◽

Integrated Approach ◽

Binding Pocket ◽

Loss Of Function ◽

Tertiary Structures ◽

Ecdysteroid Biosynthesis ◽

17Β Estradiol ◽

Unique Amino Acid

Ecdysteroids are the principal steroid hormones essential for insect development and physiology. In the last 18 years, several enzymes responsible for ecdysteroid biosynthesis encoded by Halloween genes were identified and genetically and biochemically characterized. However, the tertiary structures of these proteins have not yet been characterized. Here, we report the results of an integrated series of in silico, in vitro, and in vivo analyses of the Halloween GST protein Noppera-bo (Nobo). We determined crystal structures of Drosophila melanogaster Nobo (DmNobo) complexed with GSH and 17β-estradiol, a DmNobo inhibitor. 17β-Estradiol almost fully occupied the putative ligand-binding pocket and a prominent hydrogen bond formed between 17β-estradiol and Asp-113 of DmNobo. We found that Asp-113 is essential for 17β-estradiol–mediated inhibition of DmNobo enzymatic activity, as 17β-estradiol did not inhibit and physically interacted less with the D113A DmNobo variant. Asp-113 is highly conserved among Nobo proteins, but not among other GSTs, implying that this residue is important for endogenous Nobo function. Indeed, a homozygous nobo allele with the D113A substitution exhibited embryonic lethality and an undifferentiated cuticle structure, a phenocopy of complete loss-of-function nobo homozygotes. These results suggest that the nobo family of GST proteins has acquired a unique amino acid residue that appears to be essential for binding an endogenous sterol substrate to regulate ecdysteroid biosynthesis. To the best of our knowledge, ours is the first study describing the structural characteristics of insect steroidogenic Halloween proteins. Our findings provide insights relevant for applied entomology to develop insecticides that specifically inhibit ecdysteroid biosynthesis.

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