Search

Uterine leiomyosarcoma (LMS) is a rare but deadly disease. Due to poor understanding of the molecular and genetic causes of the disease, the diagnosis of LMS has been based primarily on histology. Nuclear atypia is one of hallmarks in LMS, however, it also occurs in 2 clinically benign variants, including smooth muscle tumors with fumarate hydratase alteration (SMT‐FH) and leiomyoma with bizarre nuclei (LM‐BN). In addition to nuclear atypia, many well recognized biomarkers used for LMS are also frequently overexpressed in LM‐BN, and the histogenesis and molecular natures for LM‐BN and LMS remain largely unknown. To characterize the molecular profiling of LMS, SMT‐FH, and LM‐BN, we performed integrated comprehensive genomic profiling including whole‐genome sequencing (WGS) and RNA sequencing and genomic microarray analyses to assess genome‐wide copy number alterations (CNAs) and immunohistochemistry (IHC) in all 3 tumor types. We found that both LM‐BN and LMS showed genomic instability and harbored extensive CNAs throughout the whole genome. By contrast, the SMT‐FH presented its characteristic 1q43‐44 deletions in all cases tested, with minimal CNAs in the rest of genomic regions. Further analyses revealed that LMS and LM‐BN groups showed similar patterns of CNAs that are tended to cluster together and separated from the SMT‐FH group. The integrated molecular profiling enabled the detection of novel and traditional biomarkers and showed excellent discrimination between LM‐BN and LMS. Our study suggests that LM‐BN, despite having similar nuclear atypia to SMT‐FH, showed similar genomic instability but distinct genomic alterations with its malignant counterpart of LMS. The integrated molecular profiling is of clinical importance in characterizing these rare uterine smooth muscle tumors., Leiomyosarcoma and 2 benign mimics (leiomyoma with FH alteration [SMT‐FH] and leiomyoma with bizarre nuclei [LM‐BN]) have characteristic nuclear atypia and often cause diagnostic challenge. LM‐BN and LMS are highly genomically unstable and harbor extensive CNAs in the genome, whereas SMT‐FH shows simple 1q43‐44 deletions. Integrated molecular profiling enabled the detection of novel and traditional biomarkers and showed excellent discrimination between LM‐BN and LMS.

Does the aggregation of M2 macrophages affect the expression of gene associated with retinoid-interferon-induced mortality 19 (GRIM-19) in adenomyosis?Endometrial tissues were collected from patients with (n = 15) and without (n = 15) adenomyosis. Tissues were analysed for GRIM-19 and toll-like receptor 4 (TLR4) expression by immunohistochemistry and western blotting. Apoptosis was analysed by TdT (terminal deoxynucleotidyl transferase)-mediated dUDP nick-end labelling (TUNEL) assay. Human endometrial stromal cells (HESC) were transfected with GRIM-19 small interfering RNA (SiRNA) to knockdown GRIM-19 expression. The HESC were co-cultured with M2 macrophages to detect the influence of M2 macrophages in HESC cells. Analyses included GRIM-19, caspase-3 and TLR4 expression by western blotting, and GRIM-19 and TLR4 by quantitative real-time polymerase chain reaction. Apoptosis was measured by flow cytometry and TUNEL assay. Cell proliferation (Cell Counting Kit-8 assay) and migration assays were carried out.The expression of GRIM-19 was significantly lower in adenomyosis lesions compared with controls (P0.001). Deficiency of GRIM-19 induced by siRNA decreased apoptosis and increased proliferation and migration in HESC. A significant decrease in GRIM-19 expression occurred in HESC after co-culture with M2 macrophages (P = 0.018). After co-culture with M2 macrophage, apoptosis decreased and proliferation and cell invasion in HESC increased. Protein (P = 0.006) and mRNA (P = 0.013) expression of TLR4 in HESC also reduced after this co-culture. Up-regulation of GRIM-19 occurred in HESC treated with the activator TLR4 (P = 0.016). Up-regulation of GRIM-19 was significantly reversed in cells treated with the TLR4 inhibitor (P = 0.011).M2 macrophages may be involved in regulating the expression of GRIM-19 partly through the TLR4 signalling axis in adenomyosis.

As a commonly used phthalate compound, di(n-butyl) phthalate (DBP) is an emerging group of polyvinyl chloride plasticizers. The acute toxicity of DBP has been extensively studied using the aquatic indicator organism, Daphnia magna. However, little is known about chronic and transgenerational toxicity of DBP. In this study, acute LC50 values were 3.04 mg/L (24 h) and 2.55 mg/L (48 h). Chronic toxicity tests in the case of maternal exposure to DBP revealed that DBP had negligible effects on growth and reproduction of F3 generation of D. magna, although the growth rate of body length and the intrinsic rate of increase were prominently reduced, to a pretty small extent. At specific concentrations, DBP generated beneficial effects on the parental generation of D. magna and no obvious impacts on the F1 generation. This study showed that maternal exposure to DBP did not cause any transgenerational effects on D. magna.

Di-(2-ethylhexyl) phthalate (DEHP) is a plasticizer used in the polyvinyl chloride industry worldwide. DEHP exists in the aquatic environments for decades. However, the toxicological effects of DEHP to aquatic organisms have not been adequately researched. We investigated acute toxicity, oxidative damage, antioxidant enzyme activities, and gene expression patterns of antioxidant enzymes in juvenile and adult Daphnia magna exposed to DEHP. We found that the median lethal concentrations (LC50) of DEHP for juveniles exposed for 24 and 48 h were 0.83 and 0.56 mg L−1, respectively. The LC50 of DEHP in adults exposed for 24 and 48 h were 0.48 and 0.35 mg L−1. Daphnia magna that was exposed to DEHP had increased malondialdehyde levels for 24 h and lower total antioxidant capacity compared with the control. Activity levels of antioxidant enzymes superoxide dismutase and phase II detoxifying enzyme glutathione S-transferases were significantly higher upon initial exposure for 24 h, and enzyme activity was then diminished at high concentrations and prolonged exposure for 48 h. Gene expression levels of cat and gst were notably reduced or increased upon DEHP exposure. These findings suggest that DEHP can cause biochemical and physiological effects in juvenile and adult D. magna by inhibiting enzymes, an increase in lipid peroxidation levels and changes both transcription levels of enzymes (cat, gst). On the whole, juveniles and adults both responded similarly to DEHP. Our findings will contribute to the understanding of toxic mechanisms in phthalate esters and the evaluation of environmental risks in aquatic ecosystems.

To study the effect of indoleamine 2,3-dioxygenase (IDO) expressed in HTR-8/SVneo cells on NKG2D and NKp46 expression and cytotoxicity of decidual NK (dNK) and peripheral NK (pNK) cells.CD56(+) dNK and pNK cells purified were cultured with HTR-8/SVneo cell conditioned medium (CM), 1-MT+HTR-8/SVneo cell CM, and complete RPMI 1640 medium (negative control) in vitro. The mRNA and protein expression of NKG2D and NKp46 in NK cells were then assessed by qRT-PCR and flow cytometry, respectively. Their cytotoxicity was evaluated with LDH assays, and TNF-α secretion was analyzed by ELISA.For dNK cells, the mRNA and protein expression of NKp46 as well as NKG2D did not differ significantly among the three groups (P0.05), whereas for pNK cells, the expression level was significantly decreased in HTR-8/SVneo cell CM group than the other two groups (P0.01). Peripheral NK cells cultured with HTR-8/SVneo cell CM showed reduced cytotoxicity and TNF-α secretion than the other two groups (P0.01), although there were no significant differences among three groups for dNK cells (P0.05).IDO expressed by HTR-8/SVneo cells can down-regulate NKp46 and NKG2D expression and reduce cytotoxicity in pNK cells, and may contribute to keep dNK cytotoxicity at a low level, suggesting an important role for IDO in the maintenance of normal pregnancy.

Uterine leiomyomas (ULM) are histologically and molecularly heterogeneous and clinically they grow at vastly different rates. Several driver gene mutations have been identified in ULM, including MED12 mutations, HMGA2 overexpression, and biallelic FH inactivation. ULM with different driver mutant genes may use different molecular pathways, but currently no clear correlation between gene mutations and growth related pathways has been established. To better define this relationship, we collected ULM with MED12 (n = 25), HMGA2 (n = 15), and FH (n = 27) mutations and examined the sex steroid hormone, cell cycle, and AKT pathway genes by immunohistochemistry. While ER and PR were highly expressed in all types of ULM, FH ULM showed lower ER expression and higher PR expression. HMGA2 tumors had significantly higher levels of AKT signaling and mitogenic activity than other ULM types. HMGA2 activated AKT signaling through upregulation of IGF2BP2. Silencing HMGA2 in ULM cells resulted in downregulation of AKT and upregulation of p16 and p21, which eventually led to cell senescence. HMGA2 overexpression in ULM is not only related to tumor development but also plays a role in controlling cellular proliferation through the AKT pathway.

Objectives To localize indoleamine 2,3-dioxygenase (IDO) mRNA and protein and to undertake a functional study at the first trimester fetal–maternal interface in order to determine whether the distribution and function of IDO are related to recurrent spontaneous abortion (RSA). Methods Women undergoing legal pregnancy termination and women with RSA participated in this prospective study. Immunohistochemistry and real-time reverse transcription–polymerase chain reaction were used to analyse levels of IDO protein and mRNA in placenta, decidua and HTR-8/SVneo cells. Culture medium collected from trophoblast villous explant or HTR-8/SVneo cell cultures was used to measure IDO activity in response to interferon (IFN)-γ treatment. Results A total of 40 healthy women and 26 women with RSA provided samples of placenta and decidua. For normal pregnancies, IDO protein and mRNA was identified in placental trophoblasts, invasive extravillous trophoblasts and decidual glandular epithelium. IFN-γ significantly increased IDO activity in trophoblast villous explants and HTR-8/SVneo cells. Levels of IDO protein and mRNA in the placenta and decidua from normal pregnancies were significantly higher than in those from RSA. Conclusions Decreased levels of IDO protein and mRNA in the placenta and decidua from RSA suggest an important role for IDO in the maintenance of normal pregnancy.