Increased expression of neurogenic factors in uterine fibroids

2019 ◽  
Author(s):  
Alice Luddi ◽  
Camilla Marrocco ◽  
Laura Governini ◽  
Bianca Semplici ◽  
Valentina Pavone ◽  
...  
Keyword(s):  
Uterine Fibroids ◽  
Reproductive Age ◽  
In Vivo Studies ◽  
Pcr Analysis ◽  
Benign Tumors ◽  
Endometrial Stromal Cells ◽  
Eutopic Endometrium ◽  
Qrt Pcr ◽  
Fertility Sparing

Abstract STUDY QUESTION Are selective markers for the neuronal differentiation such as microtubule-associated protein 2 (MAP-2) and synaptophysin (SYP) as well as the nerve growth factor (NGF) expressed by fibroids, myometrium and eutopic endometrium? SUMMARY ANSWER Neuronal markers NGF, MAP-2 and SYP are highly expressed in fibroids compared with matched myometrium, and this neurogenic pathway is upregulated by tumor necrosis factor (TNF) alpha in cultured smooth muscle cells (SMCs). WHAT IS KNOWN ALREADY Uterine fibroids or leiomyomas are the most common benign tumors, accounting for approximately one-third of hysterectomies. The present trend is to improve the medical treatment avoiding surgery, also for fertility sparing; hence, the pathogenic mechanisms are investigated, aiming to develop new therapeutic strategy. STUDY DESIGN, SIZE, DURATION This laboratory-based case–control study is focused on fibroids and myometrial specimens obtained between 2015 and 2017 from 15 women of reproductive age at the proliferative phase of the menstrual cycle. Leiomyomas, matched myometrium and endometrium from each woman were analyzed. Control endometrium was obtained from women undergoing surgery for ovarian cyst (n = 15). PARTICIPANTS/MATERIALS, SETTING, METHODS qRT-PCR, western blotting and immunostaining were applied to evaluate the expression of neurogenic markers; the effects of TNF on NGF, MAP-2 and SYP expression in cultured SMCs from leiomyomas and matched myometrium were analyzed. MAIN RESULTS AND THE ROLE OF CHANCE qRT-PCR analyses using tissues from clinical patients showed that the levels of NGF, MAP-2 and SYP mRNA were significantly higher in uterine leiomyomas compared with their matched myometrium (P < 0.05), whereas only NGF was significantly increased in eutopic endometrium compared with healthy endometrium. In primary SMCs, isolated from fibroids or from the adjacent myometrium, NGF, MAP-2 and SYP mRNA expression were significantly increased by TNF treatment (P < 0.05). Finally, human endometrial stromal cells prepared from the endometrium of patients affected by uterine fibroids display higher TNF expression (P < 0.001). LIMITATIONS, REASONS FOR CAUTION qRT-PCR analysis and immunofluorescence validation are robust methods demonstrating a clear upregulation of neurogenic factors in leiomyomas, even though additional studies are needed to establish a correlation between increased neuronal gene expression and degree of pain, as well as the involvement of inflammation mediators in the development of the neurogenic unhinge. Therefore, more in vivo studies are needed to confirm the results achieved from primary cultured SMCs. WIDER IMPLICATIONS OF THE FINDINGS The increased expression of neurogenic factors in uterine fibroids and endometrium may contribute to explain the painful stimuli. Accordingly, these neurogenic pathways may represent potential therapeutic avenues to treat the fibroid-related disorders. STUDY FUNDING/COMPETING INTEREST(S) This study was supported by research grants from the University of Siena. The authors declare no conflict of interest. TRIAL REGISTRATION NUMBER N/A.

Guizhi Fuling Wan for uterine fibroids: A systematic review of in vivo studies

2019 ◽  
Vol 245 ◽  
pp. 112177 ◽  
Author(s):  
Mingdi Li ◽  
Andrew Hung ◽  
Angela Wei Hong Yang
Keyword(s):  
Systematic Review ◽  
Uterine Fibroids ◽  
In Vivo Studies

scholarly journals TMOD-31. ADAPTING ENGINEERED CELL THERAPIES TO UNDERSTAND AND OVERCOME GLIOBLASTOMA RESISTANCE USING INTEGRATED IN VIVO AND EX VIVO MODELS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi269-vi269
Author(s):  
Andrew Satterlee ◽  
Denise Dunn ◽  
Scott Floyd ◽  
Shawn Hingtgen
Keyword(s):  
Primary Tumor ◽  
Ex Vivo ◽  
Tumor Burden ◽  
In Vivo Studies ◽  
Pcr Analysis ◽  
Tissue Slices ◽  
Live Animal ◽  
Invasive Tumor ◽  
Recurrent Tumors

Abstract Genetically engineered neural stem cells (NSCs) are a promising therapy for the highly aggressive brain cancer glioblastoma (GBM), yet treatment durability remains a major challenge. We sought to define the events that contribute to dynamic adaption of GBM during NSC treatment and develop strategies to convert initial tumor kill into sustained GBM suppression. Using a unique hybrid tumor model treated with human skin-derived induced NSCs (iNSCs) releasing the pro-apoptotic agent TRAIL, we investigated how spatial distribution of tumor and iNSCs affects GBM adaption throughout recurrence. Serial bioluminescent imaging (BLI) was used to track tumor volumes in vivo, while a subset of mice were sacrificed 6, 13, and 20 days post-treatment to harvest brains and generate living ex vivo tissue slices. Live animal imaging showed iNSC-TRAIL treatment rapidly decreased tumor volumes when delivered into the primary tumor mass; however, minimal impact on tumor growth was observed when cells were delivered into distal regions of the brain. In contrast, high-resolution imaging of living brain sections showed extensive impacts of iNSC-TRAIL therapy that could not be visualized with BLI. The living slices showed iNSC-TRAIL treatment into the primary tumor decreased the solid, but not the invasive, tumor burden. Treatment into the lateral ventricles did impact tumor kill and was more effective at treating the invasive tumor burden and maintaining inhibition than treatment into the contralateral parenchyma. We next utilized the living tissue slices to explore the sensitivity of the recurrent tumors to TRAIL. When therapy was applied to slices harboring recurrent tumor, treatment again significantly reduced tumor volumes, suggesting that tumors had not acquired TRAIL resistance. These results informed an additional in vivo survival study and subsequent PCR analysis of untreated and recurrent tumors, and combine the fidelity of in vivo studies with the speed and spatial resolution of living brain slice technology.

scholarly journals Dickkopf-1 contributes to hepatocellular carcinoma tumorigenesis by activating the Wnt/β-catenin signaling pathway

2019 ◽  
Vol 4 (1) ◽  
Author(s):  
Rui Zhang ◽  
Hao-Ming Lin ◽  
Ruth Broering ◽  
Xiang-de Shi ◽  
Xian-huan Yu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Signaling Pathway ◽  
Tumor Formation ◽  
In Vivo Studies ◽  
Qrt Pcr ◽  
Subsequent Investigation ◽  
Dkk1 Expression ◽  
Tumor Number

AbstractDysregulation of dickkopf-related protein 1 (DKK1) expression has been reported in a variety of human cancers. We previously reported that DKK1 was upregulated in hepatocellular carcinoma (HCC). However, the role of DKK1 in HCC remains unclear. This study aimed to investigate the clinical significance and biological functions of DKK1 in HCC. The expression of DKK1 was examined in cirrhotic and HCC tissues by immunohistochemistry and quantitative real-time polymerase chain reaction (qRT-PCR). DKK1 was silenced or overexpressed in HCC cell lines, and in vitro and in vivo studies were performed. Immunohistochemistry revealed that DKK1 was weakly expressed in cirrhotic tissues (8/22, 36.4%) but upregulated in HCC tissues (48/53, 90.6%, cohort 1). Significant upregulation of DKK1 was observed in 57.6% (19/33, cohort 2) of HCC tissues by qRT-PCR, and the expression of DKK1 was associated with tumor size (P = 0.024) and tumor number (P = 0.019). Genetic depletion of DKK1 impaired the proliferation, colony-forming ability, invasion, and tumor formation of HCC cells (HepG2 and HUH-7). Conversely, forced expression of DKK1 increased the proliferation, colony-forming ability, and invasion of HepG2 and HUH-7 cells in vitro and enhanced tumor formation in vivo. Subsequent investigation revealed that the DKK1-mediated proliferation and tumorigenicity of HepG2 and HUH-7 cells is dependent on the Wnt/β-catenin signaling pathway. These findings indicate that DKK1 plays an oncogenic role in HCC by activating the Wnt/β-catenin signaling pathway.

scholarly journals Myomectomy during cesarean section

2021 ◽  
Vol 43 (3-4) ◽  
pp. 73-80
Author(s):  
Radmila Sparić ◽  
Đina Tomašević ◽  
Mladen Anđić ◽  
Miljan Pupovac ◽  
Aleksandra Pavić ◽  
...  
Keyword(s):  
Caesarean Section ◽  
Cesarean Section ◽  
Perioperative Complications ◽  
Uterine Fibroids ◽  
Surgical Site Infections ◽  
Reproductive Age ◽  
Benign Tumors ◽  
Uterine Torsion ◽  
Perioperative Bleeding ◽  
Increased Risk

Myomas (fibroids, leiomyomas) are the most common benign tumors of genital organs in women of reproductive age and represent a significant problem in women's health care. The frequency of cesarean section is higher in women with uterine fibroids. Absolute indications for myomectomy during caesarean section are: fibroids that prevent hysterotomy during caesarean section, impede uterine incision suture, hamper safe fetal extraction and cause uterine torsion. Relative indications for myomectomy during caesarean section are: subserous and pedunculated fibroids, anterior uterine wall fibroids, fibroids that can cause immediate perioperative, and puerperal complications, the patient's desire, fibroids that can cause complications in subsequent pregnancies, and fibroids that can be enucleated without additional hysterotomy. Myomectomy during caesarean section is a complex surgical procedure, associated with the possibility of considerable complications, and defining their actual frequency and risk factors for their occurrence requires further research. Myomectomy during caesarean section is associated with an increased risk of perioperative bleeding. Other perioperative complications of myomectomy during cesarean section are: disseminated intravascular coagulation, paralytic ileus, surgical site infections, sepsis, postoperative febrile morbidity, increased incidence of blood transfusions, and prolonged hospitalization.

scholarly journals SCIDOT-20. ADAPTING ENGINEERED CELL THERAPIES TO UNDERSTAND AND OVERCOME GLIOBLASTOMA RESISTANCE USING INTEGRATED IN VIVO AND EX VIVO MODELS

2019 ◽  
Vol 21 (Supplement_6) ◽  
pp. vi275-vi275
Author(s):  
Andrew Satterlee ◽  
Denise Dunn ◽  
Scott Floyd ◽  
Shawn Hingtgen
Keyword(s):  
Primary Tumor ◽  
Ex Vivo ◽  
Tumor Burden ◽  
In Vivo Studies ◽  
Pcr Analysis ◽  
Tissue Slices ◽  
Live Animal ◽  
Invasive Tumor ◽  
Recurrent Tumors

Abstract Genetically engineered neural stem cells (NSCs) are a promising therapy for the highly aggressive brain cancer glioblastoma (GBM), yet treatment durability remains a major challenge. We sought to define the events that contribute to dynamic adaption of GBM during NSC treatment and develop strategies to convert initial tumor kill into sustained GBM suppression. Using a unique hybrid tumor model treated with human skin-derived induced NSCs (iNSCs) releasing the pro-apoptotic agent TRAIL, we investigated how spatial distribution of tumor and iNSCs affects GBM adaption throughout recurrence. Serial bioluminescent imaging (BLI) was used to track tumor volumes in vivo, while a subset of mice were sacrificed 6, 13, and 20 days post-treatment to harvest brains and generate living ex vivo tissue slices. Live animal imaging showed iNSC-TRAIL treatment rapidly decreased tumor volumes when delivered into the primary tumor mass; however, minimal impact on tumor growth was observed when cells were delivered into distal regions of the brain. In contrast, high-resolution imaging of living brain sections showed extensive impacts of iNSC-TRAIL therapy that could not be visualized with BLI. The living slices showed iNSC-TRAIL treatment into the primary tumor decreased the solid, but not the invasive, tumor burden. Treatment into the lateral ventricles did impact tumor kill and was more effective at treating the invasive tumor burden and maintaining inhibition than treatment into the contralateral parenchyma. We next utilized the living tissue slices to explore the sensitivity of the recurrent tumors to TRAIL. When therapy was applied to slices harboring recurrent tumor, treatment again significantly reduced tumor volumes, suggesting tumors had not acquired TRAIL resistance. These results informed an additional in vivo survival study and subsequent PCR analysis of untreated and recurrent tumors, and combine the fidelity of in vivo studies with the speed and spatial resolution of living brain slice technology.

scholarly journals Role and Regulation of the Serum- and Glucocorticoid-Regulated Kinase 1 in Fertile and Infertile Human Endometrium

Endocrinology ◽  
2007 ◽  
Vol 148 (10) ◽  
pp. 5020-5029 ◽  
Author(s):  
Fakhera Feroze-Zaidi ◽  
Luca Fusi ◽  
Masashi Takano ◽  
Jenny Higham ◽  
Madhuri S. Salker ◽  
...  
Keyword(s):  
Marker Gene ◽  
Human Endometrium ◽  
Pcr Analysis ◽  
Luminal Epithelium ◽  
Uterine Receptivity ◽  
Forkhead Transcription Factor ◽  
Endometrial Stromal Cells ◽  
Stromal Compartment ◽  
Infertile Women

Using cDNA microarray analysis, we identified SGK1 (serum- and glucocorticoid-regulated kinase 1) as a gene aberrantly expressed in midsecretory endometrium of women with unexplained infertility. SGK1 is a serine/threonine kinase involved primarily in epithelial ion transport and cell survival responses. Real-time quantitative PCR analysis of a larger, independent sample set timed to coincide with the period of uterine receptivity confirmed increased expression of SGK1 transcripts in infertile women compared with fertile controls. We further demonstrate that SGK1 expression is regulated by progesterone in human endometrium in vivo as well as in explant cultures. During the midsecretory phase of the cycle, SGK1 mRNA and protein were predominantly but not exclusively expressed in the luminal epithelium, and expression in this cellular compartment was higher in infertile women. In the stromal compartment, SGK1 expression was largely confined to decidualizing cells adjacent to the luminal epithelium. In primary culture, SGK1 was induced and phosphorylated upon decidualization of endometrial stromal cells in response to 8-bromo-cAMP and progestin treatment. Moreover, overexpression of SGK1 in decidualizing cells enhanced phosphorylation and cytoplasmic translocation of the forkhead transcription factor FOXO1 and inhibited the expression of PRL, a major decidual marker gene. Conversely, knockdown of endogenous SGK1 by small interfering RNA increased nuclear FOXO1 levels and enhanced PRL expression. The observation that SGK1 targets FOXO1 in differentiating human endometrium, together with its distinct temporal and spatial expression pattern and increased expression in infertile patients, suggest a major role for this kinase in early pregnancy events.

scholarly journals Down-Regulating Hexokinase 2 Inhibits Proliferation of Endometrial Stromal Cells Through a Noncanonical Pathway Involving Phosphorylated-STAT1 in Endometriosis

2021 ◽  
Author(s):  
Shuhui Hou ◽  
Shating Lei ◽  
Haiyan Peng ◽  
Lichun Weng ◽  
Siji Lv ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Cell Function ◽  
Malignant Tumors ◽  
Reproductive Age ◽  
Endometrial Stromal Cells ◽  
Hexokinase 2 ◽  
Proliferation Capacity ◽  
Glycolysis Pathway ◽  
Elevated Expression

Abstract Background: Endometriosis is a benign gynecologic disease that causes chronic pelvic pain, dysmenorrhea and infertility and shares several characteristics with malignant tumors, afflicting women of reproductive age. Hexokinase 2 (HK2) plays a pivotal role as the first rate-limiting enzyme in the metabolic glycolysis pathway, and its abnormal elevation in tumors is associated with tumor genesis and metastasis. However, the expression and role of HK2 in endometriosis remain unclear.Methods: We sequenced the primary endometrial stromal cells from patients with endometrioma and adopted immunohistochemistry, quantitative real-time PCR and western blot to determine the expression of HK2. Then wound healing assays, cell invasion assays, cell proliferation assays were performed to explore the functions of HK2 in endometrial stromal cells. Furthermore, mice models of endometriosis were recruited to observe the effects of HK2 inhibitors in vivo. Lastly, glycolysis metabolism detection and transcriptome sequencing were carried out in HK2-knockdown endometrial stromal cells to analyze the mechanism of HK2 affecting cell function.Results: Endometriotic stromal cells displayed active glycolysis metabolism and elevated expression of HK2. Downregulating HK2 reduced the migration, invasion and proliferation capacity of endometrial stromal cells. Knockdown of HK2 induced upregulation of signal transducer and activator of transcription 1 (STAT1) and their phosphorylation to attenuate the proliferation of endometrial stromal cells.Conclusions: HK2 was associated with the migration, invasion and proliferation of endometrial stromal cells, which might provide new insights into the pathogenesis and treatment of endometriosis.

scholarly journals High levels of ca125 in a patient with large uterine fibroid.

2020 ◽  
Vol 19 (4) ◽  
pp. 167-170
Author(s):  
Konstantinos Zacharis ◽  
Konstantinos Dafopoulos
Keyword(s):  
Abdominal Mass ◽  
Female Genital Tract ◽  
Uterine Fibroids ◽  
Reproductive Age ◽  
Benign Tumors ◽  
Caucasian Woman ◽  
Cancer Antigen 125 ◽  
Cancer Antigen ◽  
Present Case Report ◽  
Female Genital

Uterine leiomyomas, also known as uterine fibroids, are the most common benign tumors of the female genital tract and affect 60 to 80% of women at their reproductive age. Although elevated tumor markers may be measured in benign gynaecological diseases, the association of uterine fibroids with increased levels of serum cancer antigen 125 (CA125) has not been proven to date. In the present case report we present a rare case of a 21-year-old Caucasian woman attended to our outpatient department with hypermenorrhea and pelvic discomfort that was treated for an enlarged intra-abdominal mass with an abnormally high CA-12 (777.3 U/mL).

310 COBALAMIN SUPPLEMENTATION DURING IN VITRO MATURATION IMPROVES PREIMPLANTATION DEVELOPMENT OF SHEEP EMBRYOS

2015 ◽  
Vol 27 (1) ◽  
pp. 244 ◽  
Author(s):  
F. Zacchini ◽  
P. Toschi ◽  
P. Loi ◽  
G. E. Ptak
Keyword(s):  
Blastocyst Stage ◽  
Developmental Competence ◽  
Pcr Analysis ◽  
Fisher Exact Test ◽  
Placental Development ◽  
Cleavage Rate ◽  
Fetal Survival ◽  
Qrt Pcr

Oocyte maturation process includes the establishment of proper epigenetics marks, fundamental to ensure successful pregnancy. Epigenetic maturation of the oocyte depends on one-carbon-metabolism (OCM), whose key elements (cobalamin, folate) are crucial cofactors for the transfer of methyl groups onto chromatin. Commercially available IVM-media only partially supports oocyte metabolic requirements, and thus may negatively affect epigenetic maturation. Of relevance, cobalamin, one of the OCM cofactors normally present in follicular fluid, is missing in IVM-media. We investigated if cobalamin supplementation of IVM media affects sheep oocyte developmental competence in term of subsequent embryo development, epigenetic pattern, and fetal survival. Briefly, sheep oocytes were isolated from ovaries and divided into 2 groups: an untreated control (in vitro CTR group) and oocytes in vitro matured in medium containing cobalamin at 200 p.m. (Cobalamin group). Following maturation, MII oocytes were in vitro fertilized and cultured until blastocyst stage. A cohort of blastocysts was surgically transferred to recipient ewes and then conceptuses were collected at 20 days of pregnancy. Naturally mated conceptuses were also collected (in vivo CTR group). In vitro-matured oocytes and -derived blastocysts were analysed (i) by immunofluorescence anti-5-methylcitydine and (ii) by qRT-PCR for the expression of a panel of genes (MATb, ACHY, CBS, MTHFR, DNMT1) involved in OCM pathway. Immunofluorescence results were analysed by Image J software. Decimal variables were analysed using the Mann-Whitney test, while variables were expressed as percentage with the Fisher exact test. Cobalamin exposure during IVM significantly increased (i) cleavage rate (cobalamin 130/220 v. in vitro CTR 134/191, P < 0.02) following in vitro fertilization and (ii) global DNA methylation in blastocyst-stage embryos (P < 0.05). Then, qRT-PCR analysis of a select panel of OCM genes following IVM supplemented with cobalamin revealed a downregulation of MATb, ACHY, and DNMT1 in cobalamin-treated oocytes v. in vitro CTR group (P < 0.05), while no differences were observed at blastocyst stage. Finally, we found that the survival rate at 20 days of pregnancy was comparable in in vitro-produced (IVP) embryos from cobalamin and in vitro CTR oocytes, but reduced compared to in vivo CTR (cobalamin 60%, in vitro CTR 72%, and in vivo CTR 100%). Altogether, our results showed that cobalamin supplementation in IVM medium positively affects competence of oocytes and methylation profile at the blastocyst stage, presumably through the OCM pathway. Moreover, postimplantation survival of IVP conceptus, derived from untreated and treated oocytes, was reduced compared to naturally mated ones. Further investigation will clarify whether cobalamin supplementation influences fetal and placental development of IVP conceptus.

Eplerenone Prevents Atrial Fibrosis via the TGF-β Signaling Pathway

Cardiology ◽  
2017 ◽  
Vol 138 (1) ◽  
pp. 55-62 ◽  
Author(s):  
Lili Du ◽  
Mu Qin ◽  
Yi Yi ◽  
Xiaoqing Chen ◽  
Weifeng Jiang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Western Blotting ◽  
Feedback Regulation ◽  
Underlying Mechanism ◽  
In Vivo Studies ◽  
Atrial Fibrosis ◽  
Qrt Pcr ◽  
Tgf Β1

Objectives: Eplerenone (EPL), an antagonist of the mineralocorticoid receptor, is beneficial for atrial fibrillation and atrial fibrosis. However, the underlying mechanism remains less well known. We aimed to investigate the effect of EPL on atrial fibrosis using a mouse with selective atrial fibrosis and to explore the underlying mechanisms. Methods: EPL-treated MHC-TGFcys33ser transgenic mice that have selective atrial fibrosis (Tx+EPL mice), as well as control mice, were used for in vivo studies including histological analyses, Western blotting, and qRT-PCR studies. TGF-β1-stimulated atrial fibroblasts were treated with EPL or vehicle for the in vitro studies including Western blotting and qRT-PCR studies. In addition, Smad7 siRNA was used to knock down Smad7. Results: EPL inhibited atrial fibrosis in the Tx mice. In addition, EPL suppressed the expression of fibrosis-related molecules induced by TGF-β1 in vivo and in vitro. This occurred in concert with a downregulation of Smad7 protein expression and an upregulation of p-Smad2/3 protein expression. In addition, knockdown of Smad7 by siRNA abolished the protective roles of EPL. Conclusions: EPL inhibited atrial fibrosis in Tx mice. The underlying mechanism may involve increased protein expression of Smad7, which enhances the inhibitory feedback regulation of TGF-β1/Smad signaling.

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