Expression and function of Toll-like receptors in human endometrial epithelial cell lines

In the endometrium two enzymes are known to convert estradiol to its inactive metabolite estrone: microsomal 17beta-hydroxysteroid dehydrogenase type 2 (17beta-HSD2) and peroxisomal 17beta-HSD4. In order to elucidate the particular function of each of these two different enzymes, the human endometrial epithelial cell lines HEC-1-A and RL95-2 were examined with respect to the expression of 17betaHSD isozymes. They were compared with human endometrium in vivo. Non-radioactive in situ hybridization revealed both enzymes in glandular epithelial cells of human endometrium. The two cell lines were screened for mRNA expression of 17beta-HSD 1-4 by RT-PCR and Northern blot. 17beta-HSD2 and 4 could be detected by either method, 17beta-HSD1 only by RT-PCR, 17beta-HSD3 not at all. Both cell lines were proven to have no receptor for progesterone which is known as a physiological inducer of several 17beta-HSD isozymes. To study the regulation of 17beta-HSD2 and 17betaHSD4, the concentration of fetal calf serum in the cell culture media was reduced stepwise to 0.3% by dilution with a defined serum replacement. This treatment led to an inhibition of 17beta-HSD2 mRNA expression and an increase in the mRNA expression of 17beta-HSD4. Concomitantly, distinct morphological changes were observed, such as a decrease in the number and length of microvilli and a decrease in the formation of domes on top of the monolayers. The endometrial epithelial cell lines HEC-1-A and RL95-2 represent a suitable in vitro model for further studies of the differential expression of the major endometrial HSD isozymes, independent of the effect of progesterone.

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Cell surfactomes of two endometrial epithelial cell lines that differ in their adhesiveness to embryonic cells

Molecular Reproduction and Development ◽

10.1002/mrd.22301 ◽

2014 ◽

Vol 81 (4) ◽

pp. 326-340 ◽

Cited By ~ 6

Author(s):

Sonali R. Bhagwat ◽

Tejashree Redij ◽

Kruttika Phalnikar ◽

Sumeet Nayak ◽

Swati Iyer ◽

...

Keyword(s):

Epithelial Cell ◽

Cell Lines ◽

Embryonic Cells ◽

Endometrial Epithelial Cell ◽

Epithelial Cell Lines

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The effects of sex steroid hormones and interleukin-1-beta on MUC1 expression in endometrial epithelial cell lines

Reproduction ◽

10.1530/rep.1.00883 ◽

2006 ◽

Vol 131 (4) ◽

pp. 733-742 ◽

Cited By ~ 33

Author(s):

A W Horne ◽

E-N Lalani ◽

R A Margara ◽

J O White

Keyword(s):

Epithelial Cell ◽

Cell Lines ◽

Interleukin 1 ◽

Flow Cytometric Analysis ◽

Endometrial Receptivity ◽

Muc1 Expression ◽

Interleukin 1 Beta ◽

Similar Treatment ◽

Endometrial Epithelial Cell ◽

Epithelial Cell Lines

Oestrogen, progesterone and paracrine signals from the embryo have been associated with the overall control of implantation. Changes in the expression of the heavily glycosylated transmembrane glycoprotein MUC1 mucin on the endometrial epithelium are also thought to be important for embryo attachment. Increased MUC1 expression has been correlated with elevated progesterone levels in the secretory phase of the menstrual cycle. Embryonic control of endometrial receptivity through changes in MUC1 expression could be achieved through the interleukin-1 system. Four endometrial epithelial cell lines (HEC1A, HEC1B, Ishikawa and RL592) were treated with oestrogen and progesterone (with or without interleukin-1-beta) and were subjected to immunocytochemistry and flow cytometric analysis to determine MUC1 production using MUC1 antibodies. HEC1A (oestrogen receptor (ER) and progesterone receptor (PR) positive) and HEC1B (ER positive and PR negative) were transfected with theMUC1promoter, underwent similar treatment regimes and the activity of theMUC1promoter relative to their untreated controls was determined using a chloramphenicol acetyltransferase (CAT) enzyme-linked immunoassay. Using the cell lines, we determined that endometrial MUC1 expression is up-regulated by progesterone, consistent with thein vivoincreases in MUC1 related to high progesterone levels. We also revealed that neither oestrogen, nor interleukin-1-beta, appear to modulate MUC1. Progesterone-dependent regulation of MUC1 is likely to be an important factor in determining endometrial receptivity.

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p53 Regulates Toll-Like Receptor 3 Expression and Function in Human Epithelial Cell Lines

Molecular and Cellular Biology ◽

10.1128/mcb.01202-08 ◽

2008 ◽

Vol 28 (21) ◽

pp. 6557-6567 ◽

Cited By ~ 82

Author(s):

Manabu Taura ◽

Ayaka Eguma ◽

Mary Ann Suico ◽

Tsuyoshi Shuto ◽

Tomoaki Koga ◽

...

Keyword(s):

Epithelial Cell ◽

Cell Lines ◽

Nuclear Translocation ◽

Innate Immune ◽

Microbial Pathogens ◽

Tlr3 Expression ◽

Epithelial Cell Lines ◽

Tlr3 Mrna ◽

And Function ◽

Interfering Rna

ABSTRACT Toll-like receptors (TLRs) are important sensors of microbial pathogens and mediators of innate immune responses. Although the signal transduction of TLRs is well elucidated, their basal regulation is largely unexplored. Here we show that the tumor suppressor p53 positively regulates the transcription of TLR3, a receptor for viral double-stranded RNA and poly(I-C), by binding to the p53 site in the TLR3 promoter. TLR3 expression was lower in HCT116 p53−/− cells than in HCT116 p53+/+ cells. Activation of p53 by 5-fluorouracil increased the TLR3 mRNA in epithelial cell lines with wild-type p53 but not in cell lines harboring mutant p53. Knockdown of p53 by small interfering RNA decreased the TLR3 expression. TLR3 mRNA was also lower in liver and intestine of p53−/− mice than in p53+/+ mice. Furthermore, the poly(I-C)-induced phosphorylation of IκB-α, nuclear translocation of NF-κB, and phosphorylation of interferon regulatory transcription factor 3, were drastically reduced in HCT116 p53−/− cells, indicating a dysregulation of the two signaling pathways governed by TLR3. Consequently, induction of interleukin-8 and beta interferon after poly(I-C) stimulation was impaired in HCT116 p53−/− cells. These results suggest that p53 influences TLR3 expression and function and highlight a role of p53 in innate immune response in epithelial cells.

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