Calcium concentration response to uterine ischemia: a comparison of uterine fibroid cells and adjacent normal myometrial cells.
Yang Weihong,Cheng Zhongping,Dai Hong
European journal of obstetrics, gynecology, and reproductive biology
OBJECTIVE:Uterine artery occlusion by laparoscopy (UAOL) has been used for the treatment of uterine fibroids and beneficial effects to patients have been shown in clinical studies since 2000. Fibroid cells are more susceptible to apoptosis than myometrial cells under hypoxic conditions, but the molecular mechanisms underlying this effect remain unclear. The aim of this study was to investigate the role of intracellular calcium (Ca(2+)) release mediated by Ca(2+) channel inositol 1,4,5 trisphosphate receptor1 (IP3R1)/ryanodine receptor1 (RYR1) in the apoptosis of uterine fibroid cells under hypoxia. STUDY DESIGN:We compared the expressions of IP3R1 and RYR1 in fibroid and surrounding myometrial tissue from 20 patients before UAOL. After 6h treatment under hypoxia (1% O2) with or without Ca(2+) channel blockers (heparin or/and ruthenium red), the intracellular Ca(2+) concentration, cytochrome c (Cytc) protein and cell apoptosis were determined. RESULTS:IP3R1 and RYR1 mRNA and protein levels were significantly higher in fibroid than in myometrial tissues. Under hypoxic conditions, Ca(2+) concentration in fibroid cells was significantly higher than in myometrial cells (Ca(2+): 82.69±16.92nmol/L vs 46.14±9.11nmol/L, P<0.05), and Cytc increased similarly in fibroid cells. These increases in Ca(2+) concentration, Cytc and cell apoptosis were significantly reversed by calcium blocker in fibroid cells. CONCLUSION:This study demonstrated that intracellular calcium release mediated by IP3R1/RYR1 could induce apoptosis in uterine fibroid cells under hypoxic conditions, and was responsible for the susceptibility to apoptosis of fibroid cells under UAOL.
10.1016/j.ejogrb.2013.12.013
Estrogen-mediated activation of fibroblasts and its effects on the fibroid cell proliferation.
Luo Ning,Guan Qiyu,Zheng Lihua,Qu Xiaoyan,Dai Hong,Cheng Zhongping
Translational research : the journal of laboratory and clinical medicine
In this study, we explored the role of estrogen-mediated activation of stromal fibroblasts in the pathogenesis of uterine fibroid in patients. We isolated uterine fibroids and surrounding smooth muscle from patients and separated fibroblasts using immunomagnetic beads. We also measured the expression levels of estrogen receptors in fibroblasts and examined cell proliferation, expressions of fibroblast activation protein (FAP), extracellular matrix (ECM) (fibronectin, laminin, collagen I), growth factors (transforming growth factor-β, insulin-like growth factor-1), and cell proliferation pathway stimulated by estrogen. We also silenced the expression of FAP by RNA interference and analyzed the expression levels of these markers before and after E2 stimulation. Finally, we also investigated the effect of activated fibroblast supernatant on cell proliferation of fibroblasts, smooth muscle cells, and fibroid cells. We found that fibroblasts in uterine fibroid were activated, and the expression levels of estrogen receptors from fibroid cells were higher than those from smooth muscle cells. After estrogen stimulation, the proliferation activity of fibroblast was enhanced, and the expression of FAP, ECM, and growth factors was increased; the signaling pathway involved in cell proliferation was also activated. Interestingly, the activated fibroblast supernatant stimulation can promote cell proliferation. Silencing of FAP expression could inhibit the E2-mediated biological effects. In conclusion, estrogen promotes proliferation of uterine fibroids through the activation of fibroblasts, thus, activated fibroblasts may play an important role in the pathogenesis of uterine fibroids, which could be targeted in future for the treatment of uterine fibroid.
10.1016/j.trsl.2013.11.008
Long-term efficacy and quality of life associated with laparoscopic bilateral uterine artery occlusion plus partial resection of symptomatic adenomyosis.
Liu Mingmin,Cheng Zhongping,Dai Hong,Qu Xiaoyan,Kang Le
European journal of obstetrics, gynecology, and reproductive biology
OBJECTIVE:To assess the long-term efficacy and quality of life associated with laparoscopic bilateral uterine artery occlusion plus partial resection of symptomatic adenomyosis. STUDY DESIGN:A total of 182 eligible patients with symptomatic adenomyosis were treated by laparoscopic bilateral uterine artery occlusion plus partial resection of adenomyosis from July 2003 to July 2009. Menstrual blood loss was measured using a pictorial blood loss assessment chart. Pain intensity during menstruation was evaluated on a 10-point visual analog scale (VAS). Health-related quality of life was measured using the WHOQOL-BREF. RESULTS:A total of 179 patients with 3 years follow-up were enrolled in this retrospective study. No severe complications were noted during the surgical procedure or follow-up period. The mean postoperative dysmenorrhea and menorrhagia scores were significantly improved (all p<0.01) at 3, 12 and 36 months postoperatively, compared with preoperative scores. The volume of the uterus was continuously reduced at 3, 6, 12 and 36 months postoperatively, and had shrunk by 58.3% at 36 months after surgery, compared with the preoperative volume. Notably, only 1.7% (3/179) of patients had received a hysterectomy at 36 months follow-up. In addition, patient's health-related quality of life scores were significantly increased (p<0.01) compared with preoperative scores. CONCLUSION:Laparoscopic bilateral uterine artery occlusion plus partial resection of symptomatic adenomyosis is effective. There was a very low recurrence rate detected by ultrasound at 36 months.
10.1016/j.ejogrb.2013.11.014
Knockdown of EHF inhibited the proliferation, invasion and tumorigenesis of ovarian cancer cells.
Cheng Zhongping,Guo Jing,Chen Li,Luo Ning,Yang Weihong,Qu Xiaoyan
Molecular carcinogenesis
Ovarian cancer is the most lethal gynecologic malignancy worldwide. ETS homologous factor (EHF), a member of E26 transformation specific (ETS) transcription factors, has been reported overexpressed in ovarian cancer. However, the molecular mechanism underlying the biological function of EHF in ovarian cancer is still unclear. Here, we found that EHF was elevated in ovarian cancer tissues compared with non-tumorous tissues. Moreover, high EHF expression level was correlated with short survival time of patients with ovarian cancer. Knockdown of EHF in ovarian cancer cells, SKOV3 and OVCAR3, significantly inhibited cell proliferation and increased cells population in G1 phase. The proteins promoting cell cycles (Cyclin B1, Cyclin D1, and PCNA) were down-regulated and the protein negatively regulating cell cycle progression (P21) was up-regulated after EHF knockdown. Moreover, inhibition of EHF in ovarian cancer cells dramatically induced cell apoptosis, but impaired cell adhesion and cell invasion. Furthermore, phosphorylation levels of ERK and AKT were notably reduced in EHF knockdown cells. Finally, in vivo data showed that knockdown of EHF inhibited tumor growth in nude mice. Our data indicates that EHF could be a potential prognosis marker for ovarian cancer and work as an oncogene by targeting ERK and AKT signaling, which can serve as a new target for ovarian cancer treatment. © 2015 Wiley Periodicals, Inc.
10.1002/mc.22349
ARHGAP10, downregulated in ovarian cancer, suppresses tumorigenicity of ovarian cancer cells.
Luo N,Guo J,Chen L,Yang W,Qu X,Cheng Z
Cell death & disease
Rho GTPase-activating proteins (RhoGAPs) are implicated in the development and progression of ovarian cancer. ARHGAP10 is a member of RhoGAP proteins and inactivates Cdc42 by converting GTP-bound form to GDP-bound form. Here, we aimed to evaluate ARHGAP10 expression profile and functions in ovarian cancer. The decreased expression of ARHGAP10 was found in 77.3% (58/75) of ovarian cancer tissues, compared with their non-tumorous counterparts. Furthermore, overall survival in ovarian cancer patients with higher expression of ARHGAP10 was longer than those with lower expression. Ectopic expression of ARHGAP10 in two ovarian cancer cell lines with lower expression of ARHGAP10 (A2780 and HO-8910) dramatically suppressed cell proliferation in vitro. In nude mice, its stable overexpression significantly inhibited the tumorigenicity of A2780 cells. We further demonstrated that overexpression of ARHGAP10 significantly inhibited cell adhesion, migration and invasion, resulted in cell arrest in G1 phase of cell cycle and a significant increase of apoptosis. Moreover, ARHGAP10 interacted with Cdc42 and overexpression of ARHGAP10 inhibited the activity of Cdc42 in A2780 cells. Gene set enrichment analysis on The Cancer Genome Atlas dataset showed that KEGG cell cycle, replication and base excision repair (BER) pathways were correlatively with the ARHGAP10 expression, which was further confirmed in ovarian cancer cells by western blotting. Hence, ARHGAP10 may serve as a tumor suppressor through inactivating Cdc42, as well as inhibiting cell cycle, replication and BER pathways. Our data suggest an important role of ARHGAP10 in the molecular etiology of cancer and implicate the potential application of ARHGAP10 in cancer therapy.
10.1038/cddis.2015.401
Multicentre study to evaluate the clinical effects of laparoscopic uterine artery occlusion in combination with myomectomy to treat symptomatic uterine leiomyomas.
Yang W,Cheng Z,Yu J,Yang H,Liu Z,Ren Q,Xu L
European journal of obstetrics, gynecology, and reproductive biology
OBJECTIVE:Uterine artery occlusion is often used to treat symptomatic uterine myomas, as this is a minimally invasive approach. However, alternative methods for uterus-sparing therapy are in development. This study aimed to compare the clinical effects of laparoscopic uterine artery occlusion (LUAO) in combination with laparoscopic myomectomy (LM) with LM alone for the management of symptomatic uterine leiomyomas. STUDY DESIGN:This multicentre study was a retrospective controlled investigation. In total, 618 patients with symptomatic uterine myomas from six hospitals in Eastern China underwent LUAO+LM or LM alone between June 2011 and December 2012. Operative time, blood loss, transfusion, highest temperature, postoperative hospital stay, complications and follow-up results were compared between the two groups. RESULTS:Complete clinical data were available for 504 patients. Among these, 324 patients underwent LUAO+LM and 180 patients underwent LM alone. Mean±standard deviation blood loss, transfusion, highest peri-operative temperature and duration of hospital stay were significantly lower in the LUAO+LM group compared with the LM group (83.61±53.70ml vs 109±58.43ml, 1.85% vs 6.11%, 37.6±0.40°C vs 37.9±0.45°C and 5.11±0.62 days vs 6.10±0.83 days, respectively). The mean duration of follow-up was 38.97±5.82 months in the LUAO+LM group and 37.30±2.25 months in the LM group (p>0.05). The relief of abnormal uterine bleeding, reduction of uterine volume and recurrence of myomas were more remarkable in the LUAO+LM group than the LM group (97.22% vs 83.75%, 62.42% vs 51.83% and 3.47% vs 10.63%, respectively). CONCLUSIONS:LUAO in combination with LM was associated with higher surgical quality, greater relief of abnormal symptoms and less recurrence of myomas compared with LM alone. LUAO in combination with LM is recommended for women with symptomatic uterine myomas who wish to preserve their uterus.
10.1016/j.ejogrb.2016.05.033
Uterine-Sparing Laparoscopic Pelvic Plexus Ablation, Uterine Artery Occlusion, and Partial Adenomyomectomy for Adenomyosis.
Yang Weihong,Liu Mingmin,Liu Li,Jiang Caixia,Chen Li,Qu Xiaoyan,Cheng Zhongping
Journal of minimally invasive gynecology
STUDY OBJECTIVE:To evaluate safety, feasibility, and long-term clinical effects of adding laparoscopic pelvic plexus ablation to uterine-sparing procedures (uterine artery occlusion and partial adenomyomectomy) for adenomyosis. DESIGN:A prospective controlled study (Canadian Task Force classification II-1). SETTING:A teaching hospital. PATIENTS:A total of 112 patients with symptomatic adenomyosis were eligible for uterine-sparing laparoscopy. INTERVENTIONS:Laparoscopic pelvic plexus ablation, uterine artery occlusion, and partial adenomyomectomy. MEASUREMENTS AND MAIN RESULTS:After the exclusion of patients with malignant tumors or those lost to follow-up, 102 women underwent laparoscopic uterine artery occlusion and partial adenomyomectomy; 50 of these patients also had laparoscopic uterine pelvic plexus ablation (group A) with the remaining 52 patients serving as the control group (group B). Other than operative time (107.0 ± 15.4 vs 98.9 ± 20.2 minutes, p = .02), there were no statistical differences regarding other operative parameters between groups A and B. Relief of severe dysmenorrhea (Visual Analogue Scale score ≥ 7) at 36 months was higher in group A than in group B (100% vs 76.9%, p < .01). No patient suffered constipation or uroschesis in either group. CONCLUSION:Adding laparoscopic uterine pelvic plexus ablation to laparoscopic uterine artery occlusion and partial adenomyomectomy was more effective in relieving dysmenorrhea.
10.1016/j.jmig.2017.04.027
Berberine inhibits the LPS-induced proliferation and inflammatory response of stromal cells of adenomyosis tissues mediated by the LPS/TLR4 signaling pathway.
Liu Li,Chen Li,Jiang Caixia,Guo Jing,Xie Yan,Kang Le,Cheng Zhongping
Experimental and therapeutic medicine
A previous study by our group has demonstrated that lipopolysaccharide (LPS) induces adenomyosis through stimulating inflammatory cell proliferation and invasive growth of stromal cells via Toll-like receptor 4 (TLR4) signaling. The present study aimed to investigate the effects of berberine (BBR) on LPS-induced ectopic endometrial stromal cells (EESCs) isolated from patients with adenomyosis. The viability of EESCs treated with LPS or LPS plus BBR was detected by a cell counting kit-8 assay, and the cell cycle distribution and apoptosis were evaluated by flow cytometry. The effect of BBR on the expression of key molecules of inflammatory proliferation and invasive growth of LPS-induced EESCs was also evaluated. BBR significantly inhibited the LPS-induced proliferation of EESCs in a dose- and time-dependent manner. BBR induced cell cycle arrest in G0/G1 phase and enhanced apoptosis of LPS-induced EESCs. Furthermore, BBR inhibited the expression of interleukin (IL)-6, IL-8, transforming growth factor-β, epithelial growth factor, vascular endothelial growth factor and matrix metalloproteinase 2 in LPS-induced EESCs. To the best of our knowledge, the present study was the first to demonstrate that BBR has a protective effect on ameliorating the LPS-induced progression of adenomyosis. This result may provide a novel therapeutic strategy for the clinical treatment of the disease.
10.3892/etm.2017.5316