Myometrial Smooth Muscle Cells Research Articles

Regulation of sPLA2-IID in Human Decidua: Insights Into the Complexity of the Prostaglandin Pathway in Labor

Prostaglandins are implicated in the labor process, yet the precise role and regulation of the prostaglandin pathway remains to be elucidated. The first step in the pathway is cleavage of membrane phospholipids by phospholipase A2 (PLA2). Previous work demonstrated upregulation of secretory PLA2 (sPLA2)-IIA with labor in human myometrium, and recent evidence shows that there are numerous PLA2 isoforms. The present study investigates the potential of additional sPLA2 isoforms during pregnancy and labor. Real-time reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunohistochemistry were used to determine sPLA2 expression and localization. Results show the presence of sPLA2-IID in amnion, chorion, placenta, decidua, and myometrium. Expression of sPLA2-IID in decidua was significantly decreased in term labor compared to nonlabor patients, whereas no significant labor-associated changes were observed in other gestational tissues. Secretory PLA2-IID was localized within chorion fibroblasts, placenta trophoblasts, decidual cells, and in myometrial smooth muscle cells. In primary decidual cell cultures, interleukin (IL) 10 (IL-10) increased sPLA2-IID messenger RNA (mRNA) expression, while IL-1β had no effect on sPLA2-IID mRNA expression. In conclusion, decreased expression of sPLA2-IID in the decidua at labor indicates that it is unlikely to contribute to increased prostaglandin production during labor. However, increased expression of sPLA2-IID, induced by IL-10, suggests that sPLA2-IID may play an important anti-inflammatory role at the maternal-fetal interface. Nevertheless, precise functions of sPLA2-IID within the human uterus remain to be determined.

Differential expression of caveolin-1 in human myometrial and uterine leiomyoma smooth muscle

Uterine leiomyomas, the most common neoplasms of the female genital tract, are benign tumors of the uterus arising from the smooth muscle cells (SMCs) of the myometrium with an involvement of estrogen. Caveolin-1 (Cav-1), a major protein component in caveolae membrane lipid rafts, is down-regulated in several estrogen-related cancer cells, and overexpression of Cav-1 inhibits proliferation of cancer cells and vascular SMCs as well. Therefore, we hypothesize that Cav-1 is down-regulated in human uterine leiomyoma. Western blot using tissues from clinical patients showed that Cav-1 expression was significantly lower or undetectable in uterine leiomyoma compared with their matched myometrium (P < .001). This finding was confirmed by immunohistochemistry and confocal microscopy. The cav-1 mRNA level in uterine leiomyomas was also significantly lower as detected by reverse transcription-quantitative polymerase chain reaction analysis (P = .001). To further study the underlying mechanism, we performed primary cell culture, and found that the expression of Cav-1 remained low in cultured leiomyoma SMCs (P = .009). Serum withdrawal did not change Cav-1 expression in leiomyoma SMCs, but increased expression in myometrial SMCs (P = .006). 17-β estradiol inhibited the expression of Cav-1 protein (P = .047) and mRNA (P = .007) in leiomyoma SMCs, whereas it stimulated expression in myometrial SMCs (P = .043). 17-β estradiol, although activating the mitogen-activated protein kinase pathway in both SMCs, did not stimulate their proliferation. We conclude that human uterine leiomyomas in vitro express low levels of Cav-1, which may result from estrogen inhibition. This effect of estrogen may contribute to the pathogenesis of uterine leiomyoma. Further studies in vivo are needed to verify these results.

Transcriptomic effects of estradiol treatment on cultured human uterine smooth muscle cells

Contractility of the myometrial smooth muscle cells during the estrous cycle and pregnancy is modulated by estrogen but the temporal expression of estrogen (relative to progesterone) and the type of contraction involved are distinctly different in pregnancy and estrous. This in vitro cell culture study investigated the global gene expression profile of human uterine smooth muscle cells (hUtSMCs) following 17β-estradiol (E2) treatment. In response to E2 treatment 540 genes, many of which have not been previously described as estrogen responsive, were identified as significantly differentially expressed. These genes are involved in biological processes that include muscle contraction, cell migration and adhesion, apoptosis and phosphorylation. Evidence from this study suggests that 17β-estradiol may have effects that are contrary to an overall contraction phenotype. The hUtSMC in vitro culture system is a useful model to investigate steroid effects on smooth muscle cells and may provide additional clues as to how smooth muscle cells behave in vivo.

The role of brain natriuretic peptide in maintaining myometrial quiescence during pregnancy

Myometrial quiescence is a physiological stage of the myometrium during pregnancy. It is a period of active relaxation of the myometrial smooth muscle cells; myometrial quiescence is responsible for maintaining pregnancy. The precise mechanisms underlying myometrial quiescence have not been completely elucidated, although many mediators and cellular pathways have been described as playing a role. Fetal membranes (chorion and amnion) produce and release one or more substances that inhibit myometrial contractions, playing a central role in the maintenance of myometrial quiescence. Brain natriuretic peptide (BNP) is more potent than any other natriuretic peptide in inhibiting myometrial contractions in vitro. Brain natriuretic peptide is produced by the chorion and amnion, mainly during myometrial quiescence, and decreasing towards the end of pregnancy. Production of BNP is reduced in fetal membranes obtained from women in preterm labour. It is postulated that BNP, acting in a paracrine fashion, plays a key role in the maintaining myometrial quiescence and, therefore, controlling the duration of pregnancy. Furthermore, it is postulated that a premature decrease of BNP production by the fetal membranes may cause preterm labour and preterm birth.

709: Vitamin D regulates RANK expression in term human myometrial smooth muscle cells

709: Vitamin D regulates RANK expression in term human myometrial smooth muscle cells

Vitamin D regulates contractile profile in human uterine myometrial cells via NF-κB pathway

Infection triggers inflammation that, in turn, enhances the expression of contractile-associated factors in myometrium and increases the risk of preterm delivery. In this study, we assessed vitamin D regulation of inflammatory markers, contractile-associated factors, steroid hormone receptors, and NFκB pathway proteins in human uterine myometrial smooth muscle (UtSM) cells that were cultured in an inflammatory environment. Inflammatory environment was simulated for UtSM cells by coculturing them with monocyte lineage (THP1) cells. We measured the expression of inflammatory markers, contractile-associated factors, steroid hormone receptors, and NFκB pathway proteins in UtSM cells that were cultured with THP1 cells in the presence and absence of vitamin D by real time polymerase chain reaction and Western blot analysis. Monocytes secreted monocyte inflammatory protein-1α and -1β, interleukin (IL)-1β and 6, and tumor necrosis factor-α into the conditioned medium. In the UtSM cells that had been cocultured with THP1 cells, there was a significant (P < .05) increase in the expression of inflammatory markers IL-1β, -6, and -13 and tumor necrosis factor-α; the contractile-associated factors connexin-43, Cox-2, and prostaglandin F2α receptor; the estrogen receptor α, and progesterone receptors A and B. Vitamin D treatment of cocultures decreased (P < .05) the expression of inflammatory markers and contractile-associated factors in UtSM cells. Similarly, vitamin D decreased estrogen receptor α and progesterone receptors A-to-B ratio in UtSM cells that were cocultured with THP1 cells. In addition, vitamin D treatment significantly (P < .05) decreased monocyte-induced p-IκBα in cytosol and NFκB-p65 in the nucleus and increased IκBα in cytosol in UtSM cells. Our results suggest that vitamin D treatment decreases inflammation-induced cytokines and contractile-associated factors in the uterine myometrial smooth muscle cells through the NFκB pathway.

Paracrine activation of WNT/β-catenin pathway in uterine leiomyoma stem cells promotes tumor growth

Uterine leiomyomas are extremely common estrogen and progesterone-dependent tumors of the myometrium and cause irregular uterine bleeding, severe anemia, and recurrent pregnancy loss in 15-30% of reproductive-age women. Each leiomyoma is thought to arise from a single mutated myometrial smooth muscle stem cell. Leiomyoma side-population (LMSP) cells comprising 1% of all tumor cells and displaying tumor-initiating stem cell characteristics are essential for estrogen- and progesterone-dependent in vivo growth of tumors, although they have remarkably lower estrogen/progesterone receptor levels than mature myometrial or leiomyoma cells. However, how estrogen/progesterone regulates the growth of LMSP cells via mature neighboring cells is unknown. Here, we demonstrate a critical paracrine role of the wingless-type (WNT)/β-catenin pathway in estrogen/progesterone-dependent tumorigenesis, involving LMSP and differentiated myometrial or leiomyoma cells. Estrogen/progesterone treatment of mature myometrial cells induced expression of WNT11 and WNT16, which remained constitutively elevated in leiomyoma tissues. In LMSP cells cocultured with mature myometrial cells, estrogen-progesterone selectively induced nuclear translocation of β-catenin and induced transcriptional activity of its heterodimeric partner T-cell factor and their target gene AXIN2, leading to the proliferation of LMSP cells. This effect could be blocked by a WNT antagonist. Ectopic expression of inhibitor of β-catenin and T-cell factor 4 in LMSP cells, but not in mature leiomyoma cells, blocked the estrogen/progesterone-dependent growth of human tumors in vivo. We uncovered a paracrine role of the WNT/β-catenin pathway that enables mature myometrial or leiomyoma cells to send mitogenic signals to neighboring tissue stem cells in response to estrogen and progesterone, leading to the growth of uterine leiomyomas.

Spatial heterogeneity enhances and modulates excitability in a mathematical model of the myometrium

The muscular layer of the uterus (myometrium) undergoes profound changes in global excitability prior to parturition. Here, a mathematical model of the myocyte network is developed to investigate the hypothesis that spatial heterogeneity is essential to the transition from local to global excitation which the myometrium undergoes just prior to birth. Each myometrial smooth muscle cell is represented by an element with FitzHugh-Nagumo dynamics. The cells are coupled through resistors that represent gap junctions. Spatial heterogeneity is introduced by means of stochastic variation in coupling strengths, with parameters derived from physiological data. Numerical simulations indicate that even modest increases in the heterogeneity of the system can amplify the ability of locally applied stimuli to elicit global excitation. Moreover, in networks driven by a pacemaker cell, global oscillations of excitation are impeded in fully connected and strongly coupled networks. The ability of a locally stimulated cell or pacemaker cell to excite the network is shown to be strongly dependent on the local spatial correlation structure of the couplings. In summary, spatial heterogeneity is a key factor in enhancing and modulating global excitability.

Blockade of the endothelinA receptor prevents hypoxia induced smooth muscle cell proliferation and endothelin secretion in patients with uterine leiomyomas

Hypoxia, low oxygen conditions, has been implicated as a possible mechanism by which uterine myometrial cells undergo aberrant cell growth and differentiation into leiomyoma (fibroid) cells. Endothelin-1 (ET-1), a vasoconstrive peptide, is increased in fibroid explants in response to hypoxia and inhibits apoptosis of leiomyoma cells in vitro. The objective of this study was to determine if hypoxia induces ET-1 secretion from myometrial smooth muscle cells (mSMCs) to levels comparable to ET-1 secretion fibroid smooth muscle cells (fSMCs). Basic science research study using human tissue samples. Fibroid and myometrium tissue were excised from the uterus of women undergoing hysterectomies for uterine fibroids (n=10). Tissue was washed in phosphate-buffered saline and digested in collagenase solution for 6hrs. The resultant smooth muscle cells were grown until confluent, trypsinized and seeded at 50,000 cells/well and cultured under normoxia (6% oxygen) or hypoxia (1% oxygen) for a period of 24hrs with/without 1μM of endothelin A (ETA) receptor antagonist. ET-1 was measured from cell culture supernatant using a Quantikine ET-1 ELISA. Results were analyzed with paired student's t-test. Under normoxia fSMCs secreted more ET-1(840.9+234ng/mg/mL) compared to mSMCs (265.2+65ng/mg/mL;p=.03). Hypoxia increased ET-1 secretion from mSMCs (903.1+186ng/mg/mL) to levels comparable with normoxic fSMCs (p=.854) and levels significantly higher than mSMCs cultured at normoxia (p<.05). Blockade of the ETA receptor for 24hrs significantly decreased hypoxia induced ET-1 secretion from mSMCs (148.3+18) compared to untreated hypoxic fSMCs (p=.05). Hypoxia significantly increased mSMC proliferation (p<.05) compared to normoxic mSMCs. Blockade of the ETA receptor decreased hypoxia induced increase in mSMC proliferation (p=.05). These results indicate that hypoxia may increase mSMC proliferation through activation of the ETA receptor.

Immunolocalization of vascular endothelial growth factor, its receptors (flt1/fms, flk1/KDR, flt4) and vascular endothelial growth inhibitor in the bitch uterus during the sexual cycle

Angiogenesis is regulated by proangiogenic and antiangiogenic factors. Vascular endothelial growth factor (VEGF) is a prime proangiogenic regulator, whereas vascular endothelial growth inhibitor (VEGI) is a specific antiangiogenic cytokine. To clarify temporal changes in the localization of pro-angiogenic and anti-angiogenic factors in the uterus of normal bitches during the proestrus, estrus, diestrus and anestrus phases of the estrous cycle, the expressions of VEGF and its receptors (flt1/fms, flk1/KDR and flt4) and their correlation with VEGI were analyzed using immunohistochemistry. Uteruses were collected after ovariohysterectomy. Immunohistochemical staining was evaluated semi-quantitatively by an immunohistochemical total score consisting of the sum of the intensity and proportional scores. The results in the bitch uterus demonstrated that positive immunohistochemical staining was found exclusively in the cytoplasm and apical membrane of luminal and glandular epithelial, stromal and smooth muscle cells and nuclear staining was observed in the flt1/fms, flk4 and VEGI during proestrous and estrous. Semi-quantitative analyses revealed that the total score for VEGF in the glandular epithelial cells was significantly higher than that of luminal, endometrial stromal and myometrial smooth muscle cells during proestrous (p<0.05). The total score for flk1/KDR and flt4 in the glandular epithelium was also significantly higher than that of endometrial stromal cells during proestrous, whilst the total score for flt1/fms in the glandular epithelium was significantly higher than that of endometrial stromal cells during anestrus (p<0.05). We conclude that, in the bitch uterus, cyclic changes may be precisely regulated by the combined functions of VEGF family members, angiogenic VEGF and VEGF receptors, and the angiogenesis inhibitor VEGI.

Aberrant expression and regulation of NR2F2 and CTNNB1 in uterine fibroids

Uterine fibroids are the most common benign tumour afflicting women of reproductive age. Despite the large healthcare burden caused by fibroids, there is only limited understanding of the molecular mechanisms that drive fibroid pathophysiology. Although a large number of genes are differentially expressed in fibroids compared with myometrium, it is likely that most of these differences are a consequence of the fibroid presence and are not causal. The aim of this study was to investigate the expression and regulation of NR2F2 and CTNNB1 based on their potential causal role in uterine fibroid pathophysiology. We used real-time quantitative RT-PCR, western blotting and immunohistochemistry to describe the expression of NR2F2 and CTNNB1 in matched human uterine fibroid and myometrial tissues. Primary myometrial and fibroid smooth muscle cell cultures were treated with progesterone and/or retinoic acid (RA) and sonic hedgehog (SHH) conditioned media to investigate regulatory pathways for these proteins. We showed that NR2F2 and CTNNB1 are aberrantly expressed in fibroid tissue compared with matched myometrium, with strong blood vessel-specific localisation. Although the SHH pathway was shown to be active in myometrial and fibroid primary cultures, it did not regulate NR2F2 or CTNNB1 mRNA expression. However, progesterone and RA combined regulated NR2F2 mRNA, but not CTNNB1, in myometrial but not fibroid primary cultures. In conclusion, we demonstrate aberrant expression and regulation of NR2F2 and CTNNB1 in uterine fibroids compared with normal myometrium, consistent with the hypothesis that these factors may play a causal role uterine fibroid development.

Differentially regulated expression of neurokinin B (NKB)/NK3 receptor system in uterine leiomyomata

Are the vasoactive peptide neurokinin B (NKB) and its preferred NK3 receptor (NK3R) differentially expressed in leiomyomas compared with normal myometrium? In leiomyomas, NKB is up-regulated and delocalized, while its preferred NK3R is also differentially regulated. The expression of NKB/NK3R in the central nervous system is essential for proper function of the human reproductive axis. Additionally, this system is also widely expressed throughout the female genital tract. Leiomyomas impair fertility and are a major source of abnormal uterine bleeding. The aberrant synthesis of local factors can contribute to the pathological symptoms observed in women with leiomyomata. NKB could be one of these factors, since a vasoactive role of this peptide at a peripheral level has been observed in different systems and species, including humans. NK3R is strongly regulated by estrogens and its activation leads to nuclear translocation affecting chromatin structure and gene expression. Samples were obtained between 2006 and 2012 from 28 women of reproductive age at different stages of the menstrual cycle by hysterectomy. Leiomyomas and matched macroscopically normal myometrium from each woman were analysed in vitro. RT-PCR, quantitative real time, immunohistochemistry and in situ hybridization were used to investigate the pattern of expression of NKB/NK3R in tissue samples. Expression of the gene encoding NKB (TAC3) was up-regulated 20-fold in leiomyomas, compared with matched myometrium (P = 0.0008). In tumour tissue, not only connective cells, but also myometrial, endothelial and vascular smooth muscle cells express TAC3 mRNA. Immunoreactivity to NKB was preferentially located in the smooth muscle cell nuclei from normal myometrium in the secretory phase, unlike matched leiomyoma, which showed a predominant cytoplasmic expression pattern. In the normal myometrium, TACR3 mRNA showed variable expression throughout the menstrual phases, with samples showing strong, reduced or no amplification. In leiomyoma, TACR3 was significantly up-regulated compared with matched myometrium (P = 0.0349). This study is descriptive and although we observed clear differential regulation of the NKB/NK3R system at mRNA and immunohistochemical staining levels in leiomyoma, future functional studies are needed to determine the precise role of NKB in the myometrium in normal and pathological conditions. In addition, further analysis (e.g. in cell culture models) will be required to determine the role of NKB in the nucleus of normal smooth muscle cells, whether nuclear translocation is mediated by NK3R and the consequences of the cytoplasmic expression of NKB in tumour cells. The NKB/NK3R system dysregulation observed in leiomyoma may contribute to the pathological symptoms observed in women with leiomyomata.

Vitamin D Elicits Anti-Inflammatory Response, Inhibits Contractile-Associated Proteins, and Modulates Toll-Like Receptors in Human Myometrial Cells

Infection during pregnancy triggers inflammation, which can increase myometrial contractions and the risk of premature labor and delivery. In this study, we assessed the effects of vitamin D, an anti-inflammatory ligand on cytokines, chemokines, toll-like receptors, and contractile-associated proteins on immortalized human myometrial smooth muscle (UtSM) cells stimulated with lipopolysaccharide (LPS), a bacterial endotoxin, or interleukin (IL)-1β and measured Toll-like receptor (TLR)-10 expression in pregnant myometrial tissues. A superarray analysis revealed downregulation of the chemokines monocyte chemoattractant protein (MCP)-1, Chemokine (C-X-C motif) ligand (CXCL)-10, CXCL-11, and chemokine (C-X3-C motif) ligand (CX3CL)-1; the proinflammatory cytokines IL-13 and tumor necrosis factor (TNF)-α; the TLR-4 and -5 and triggering receptor expressed on myeloid cells (TREM)-2 and upregulation of the anti-inflammatory cytokine IL-10, as well as Toll interacting protein (TOLLIP) and TREM-1 in vitamin D-treated UtSM cells. In the presence of LPS, vitamin D caused dose-dependent decreases in the messenger RNA expression of MCP-1, IL-1β, IL-13, TNF-α, TLR-4, and TLR-5, the contractile-associated proteins connexin 43, the oxytocin receptor, and the prostaglandin receptor but caused increases in IL-10 and TLR-10 in UtSM cells. The TLR-10 expression was higher in human myometrial tissue obtained from women at term not in labor compared to labor. Vitamin D also attenuated IL-1β-induced MCP-1, IL-6, connexin 43, cyclooxygenase (COX)-2, and prostaglandin receptor expression. Western analysis showed that vitamin D decreased MCP-1, TLR-4, and connexin 43 in the presence of LPS and decreased connexin 43 in the presence of IL-1β. Our results suggest that vitamin D can potentially decrease infection-induced increases in cytokines and contractile-associated proteins in the myometrium.

Corticotrophin‐releasing hormone up regulated IL‐6, IL‐8 and MCP‐1 output in primary human myometrial smooth muscle cells via CRHR1 by activating the PKA pathway

CRH has been implicated to be involved in controlling the length of pregnancy and initiating the onset of parturition in human. Given that parturition is an inflammatory process, we hypothesized that CRH regulates the inflammation responses by controlling the production of cytokines and chemokines in human myometrium, thereby leading to parturition. Primary myometrial cells were isolated and cultured from lower segmental myometrial tissues obtained from pregnant women at term. CRH treatment caused an increase in IL‐6, IL‐8 and MCP‐1 production in myometrial cells. These effects were reversed by CRHR1 antagonist but not by CRHR2 antagonist. Knockdown of CRHR1 resulted in a loss of CRH effect. CRH stimulated active Gαs protein and resulted in an increase in cAMP production in myometrial cells. CRH‐induced production of IL‐6, IL‐8 and MCP‐1 was blocked by either adenylate cyclase (AC) inhibitor SQ22536 or PKA inhibitor H89. In conclusion, CRH acts on CRHR1 to stimulate the production of cytokine and chemokine in myometrial cells. These effects are dependent on AC/PKA signaling pathway. Our data suggest that CRH can directly stimulate myometrial cells to secret chemokines and cytokines, which could recruit leukocytes to the uterus and activation of inflammation response.

Biphasic Erk1/2 activation sequentially involving Gs and Gi signaling is required in beta3-adrenergic receptor-induced primary smooth muscle cell proliferation

The beta3 adrenergic receptor (B3-AR) reportedly induces cell proliferation, but the signaling pathways that were proposed, involving either Gs or Gi coupling, remain controversial. To further investigate the role of G protein coupling in B3-AR induced proliferation, we stimulated primary human myometrial smooth muscle cells with SAR150640 (B3-AR agonist) in the absence or presence of variable G-protein inhibitors. Specific B3-AR stimulation led to an Erk1/2 induced proliferation. We observed that the proliferative effects of B3-AR require two Erk1/2 activation peaks (the first after 3min, the second at 8h). Erk1/2 activation at 3min was mimicked by forskolin (adenylyl-cyclase activator), and was resistant to pertussis toxin (Gi inhibitor), suggesting a Gs protein signaling. This first signaling also required the downstream Gs signaling effectors PKA and Src. However, Erk1/2 activation at 8h turned out to be pertussis toxin-dependent, and PKA-independent, indicating a Gi signaling pathway in which Src and PI3K were required. The pharmacological inhibition of both the Gs and Gi pathway abolished B3-AR-induced proliferation. Altogether, these data indicate that B3-AR-induced proliferation depends on the biphasic activation of Erk1/2 sequentially induced by the Gs/PKA/Src and Gi/Src/PI3K signaling pathways.

Melatonin and the Circadian Timing of Human Parturition

Although the onset of spontaneous human parturition has long been known to occur preferentially during the nighttime and early morning hours, no convincing physiological explanation for this pattern has yet been proposed. This review focuses on the circadian timing of mammalian parturition, particularly in the human. It is proposed that differences in the phasing of parturition among different species are likely a function of opposite uterine responses to humoral cues, in particular those coding for time of day. The brain hormone melatonin fulfills many of the prerequisites to serve as a circadian signal for initiating uterine contractions that lead to human parturition. These encompass direct actions of melatonin on myometrial smooth muscle cells that are synergistic with oxytocin in facilitating greater uterine contractions at night. This may not only help to explain the nocturnal phasing of human parturition but also open new avenues for the management of term and preterm labor.

Effect of corticotrophin-releasing hormone on connexin-43 phosphorylation and gap junction intercellular communication in human myometrial smooth muscle cells

To determine the effect of human corticotrophin-releasing hormone (CRH) on the expression of connexin-43 phosphate (P-Cx43) in human myometrial smooth muscle cells (SMCs) and the function of cell gap junction intercellular communication in SMCs. Human non-conceive myometial SMCs were cultured with different concentrations of CRH (0, 5.85, 58.5, 585 and 5850 pmol/L). Western blot was used to test P-Cx43 and Cx43 non-phosphate (NP-Cx43) of protein expression. Cell scratch was used to test cell gap junction intercellular communication opening status in human myometrial SMCs. Compared with the control group, the expression of P-Cx43 was higher in the CRH groups (P<0.01), and was concentration-dependent. There was no significant difference in NPCx43 between the control group and the CRH groups (P>0.05). The transmission of cell layers in the CRH groups was higher than that in the control group (P<0.01), and as the concentration of CRH increased, the time was concentration-dependent (P<0.01). CRH can enhance the expression of P-Cx43 and the function of gap junction intercellular communication in the primary cultured myometrial SMCs.

477: Purkinje cell protein 4 (PCP4) as a regulator of myometrial smooth muscle cell function

477: Purkinje cell protein 4 (PCP4) as a regulator of myometrial smooth muscle cell function

Differential expression of G-protein-coupled estrogen receptor-30 in human myometrial and uterine leiomyoma smooth muscle

To determine differential expression of G-protein-coupled receptor 30 (GPR30) in uterine leiomyoma and its matched myometrium. GPR30 expression examined in both tissues and cultured cells. Research laboratories. Women 35 to 50 years old with uterine leiomyomas. Hysterectomy. GPR30 expression profile. Using Western blot and real-time quantitative polymerase chain reaction analyses, we found that GPR30 was highly expressed in uterine leiomyomas compared with their matched myometrium. In only three out of nine patients examined was GPR30 protein detectable by Western blot analysis in myometrial tissues, but at statistically significantly lower levels than in their leiomyomas. Confocal microscopy revealed the nuclear localization of GPR30 in leiomyoma tissues and cultured leiomyoma smooth muscle cells (SMCs). Treatment with 0.1 μM 17β-estradiol increased mRNA expression of GPR30 in leiomyoma SMCs but decreased expression in myometrial SMCs. Treatment with G-1, a GPR30 agonist, stimulated phosphorylation of p44/42 mitogen-activated protein kinase (MAPK) in both SMC types. PD98059, the MEK inhibitor, completely inhibited G-1-induced phosphorylation of p44/42 in myometrium SMCs, but not in SMCs from leiomyoma. GPR30 is abundantly expressed in uterine leiomyomas, likely resulting from estrogen stimulation.

M270 DIFFERENTIAL EXPRESSION OF CAVEOLIN-1 IN UTERINE LEIOMYOMA AND MYOMETRIAL SMOOTH MUSCLE CELLS: THE POTENTIAL ROLE OF ESTROGEN

International Journal of Gynecology & ObstetricsVolume 119, Issue S3 p. S619-S619 Poster presentations M270 DIFFERENTIAL EXPRESSION OF CAVEOLIN-1 IN UTERINE LEIOMYOMA AND MYOMETRIAL SMOOTH MUSCLE CELLS: THE POTENTIAL ROLE OF ESTROGEN Y. Zhu, Y. ZhuSearch for more papers by this author Y. Zhu, Y. ZhuSearch for more papers by this author First published: 22 October 2012 https://doi.org/10.1016/S0020-7292(12)61462-3AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onFacebookTwitterLinkedInRedditWechat No abstract is available for this article. Volume119, IssueS3Abstracts of XX FIGO World Congress of Gynecology and ObstetricsOctober 2012Pages S619-S619 RelatedInformation