Management of ectopic pregnancy after in vitro fertilization/intracytoplasmic sperm injection and embryo transfer: a case series and mini-review.

Background: Ectopic pregnancy (EP), reflecting a fertilized ovum implanted outside the normal uterine cavity, represents a frequent cause of morbidity and possibly mortality in women of reproductive age. Objective: To summarize the diagnosis and treatment of EP after in vitro fertilization/intracytoplasmic sperm injection and embryo transfer (IVF/ICSI-ET). Methods: The medical records of patients who were diagnosed with EP after embryo transfer from 2017 to 2019, in a tertiary hospital were reviewed. Results: Of the 24 cases analyzed, 21 (87.5%) had fallopian tube involvement, while 2 (8.3%) and 1 (4.2%) had cornual and cervical pregnancies, respectively. Clinical manifestations included vaginal bleeding (58.3%) and lower abdominal pain (16.7%); 9 (42.9%) cases had no symptoms. One cornual pregnancy was misdiagnosed as acute appendicitis and later correctly diagnosed by laparoscopic exploration. There were 2 cases of multiple-site EP and 2 of heterotopic pregnancy, including one with an intrauterine pregnancy with double chorionic and four amniotic sacs and right tubal ampullary pregnancy. Five of the 21 cases with fallopian tube involvement received conservative treatment, while the remaining 16 underwent surgeries, including laparoscopic ipsilateral salpingostomy and ipsilateral salpingectomy. Discussion: Ectopic pregnancy after embryo transfer, mainly involving the fallopian tube, is very complex and is with diverse manifestations. Even with the pregnancy sac observed in the uterus, the pelvic cavity should be scanned thoroughly after embryo transfer.

Akkermansia muciniphila and its outer membrane protein Amuc_1100 prevent high-fat diet-induced nonalcoholic fatty liver disease in mice.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease. A. muciniphila and its outer membrane protein Amuc_1100 ameliorate metabolic disorders, enteritis, depression, and other diseases in mice. The NAFLD mouse model was established by feeding a high-fat diet (HFD) for 10 weeks. To assess the effect of A. muciniphila and Amuc_1100 on NAFLD, we used atorvastatin, a common lipid-lowering drug, as a positive control. A. muciniphila and Amuc_1100 significantly reduced body weight and serum ALT and AST levels, and improved serum lipid levels in NAFLD mice, which had similar effects to Ator. In addition, A. muciniphila and Amuc_1100 decreased the concentration of LPS in the serum and upregulated the mRNA expression of the colonic tight junction proteins. In the liver, A. muciniphila and Amuc_1100 significantly reduced the mRNA expression levels of nodular receptor protein 3 (NLRP3) and Toll-like receptor 4 (TLR4)/nuclear factor κB (NF-κB), and the protein and mRNA expression levels inflammatory cytokines. At the genus level, Amuc_1100 treatment significantly reduced the abundance of Coriobacteriaceae_UCG-002 produced by the HFD. The abundances of Blautia, norank_f__Ruminococcaceae, Lachnoclostridium, GCA-900066575 and Lachnospiraceae_UCG-006 increased dramatically. Together, A. muciniphila and Amuc_1100 alleviate HFD-induced NAFLD by acting on the gut-liver axis and regulating gut microbes.

Lappaconitine sulfate inhibits proliferation and induces mitochondrial-mediated apoptosis via regulating PI3K/AKT/GSK3ß signaling pathway in HeLa cells.

Lappaconitine (LA), a diterpenoid alkaloid extracted from the root of Aconitum sinomontanum Nakai, exhibits broad pharmacological effects, including anti-tumor activity. The inhibitory effect of lappaconitine hydrochloride (LH) on HepG2 and HCT-116 cells and the toxicity of lappaconitine sulfate (LS) on HT-29, A549, and HepG2 cells have been described. But the mechanisms of LA against human cervical cancer HeLa cells still need to be clarified. This study was designed to investigate the effects and molecular mechanisms of lappaconitine sulfate (LS) on the growth inhibition and apoptosis in HeLa cells. The cell viability and proliferation were evaluated using the Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2´-deoxyuridine (EdU) assay, respectively. The cell cycle distribution and apoptosis were detected by flow cytometry analysis and 4', 6-diamidino-2-phenylindole (DAPI) staining. The mitochondrial membrane potential (MMP) was determined through the 5, 5', 6, 6'-tetrachloro-1, 1', 3, 3'-tetraethylbenzimi-dazolyl carbocyanine iodide (JC-1) staining. The cell cycle arrest-, apoptosis-, and the phosphatidylinositol-3-kinase/protein kinase B/glycogen synthase kinase 3ß (PI3K/AKT/GSK3ß) pathway-related proteins were estimated by western blot analysis. LS markedly reduced the viability and suppressed the proliferation of HeLa cells. LS induced G0/G1 cell cycle arrest through the inhibition of Cyclin D1, p-Rb, and induction of p21 and p53. Furthermore, LS triggered apoptosis through the activation of mitochondrial-mediated pathway based on decrease of Bcl-2/Bax ratio and MMP and activation of caspase-9/7/3. Additionally, LS led to constitutive downregulation of the PI3K/AKT/GSK3ß signaling pathway. Collectively, LS inhibited cell proliferation and induced apoptosis through mitochondrial-mediated pathway by suppression of the PI3K/AKT/GSK3ß signaling pathway in HeLa cells.

Characterization of the structure and properties of the isolating interfacial layer of oil-water emulsions stabilized by soy hull polysaccharide: Effect of pH changes.

The effect of pH on the microstructure and properties of the soy hull polysaccharide interfacial layer was determined. The particle size at pH 2.0 was the largest (36.7 µm), whereas the absolute ζ-potential was the smallest. The protein content was the lowest at pH 2.0 and 9.0 and peaked around pH 4.0-5.0 (77.7%). The ordered secondary protein structure content under low pH conditions was greater than that under high pH conditions and the stability of the interfacial layer was higher at high pH, whereas the emulsion viscosity decreased by two orders of magnitude between pH 2.0 and 9.0. It appears that low pH reduced the thermal stability and increased the apparent viscosity of the emulsion by increasing the structural order of the protein in the interfacial layer. These findings lay the foundation for future work to reveal the key components and characteristic structures of soy hull polysaccharide that affect interfacial stability.

Patients With Deep Ovarian Suppression Following GnRH Agonist Long Protocol May Benefit From a Modified GnRH Antagonist Protocol: A Retrospective Cohort Study.

Objective: To verify if patients with deep ovarian suppression following gonadotropin releasing hormone (GnRH) agonist long protocol may benefit from a modified GnRH antagonist protocol based on luteinizing hormone (LH) levels. Design: Retrospective cohort study. Setting: University-based hospital. Patients: 110 patients exhibited ultra-low LH levels during ovarian stimulation using GnRH agonist long protocol. Interventions: As all the embryos in the first cycle were exhausted without being pregnant, these patients proposed to undergo a second cycle of ovarian stimulation. 74 of them were treated with a modified GnRH antagonist protocol based on LH levels. Other 36 patients were still stimulated following GnRH agonist long protocol. Main Outcome Measure: The primary outcome was live birth rate (LBR). The second outcomes were biochemical pregnancy rate, clinical pregnancy rate (CPR), ongoing pregnancy rate (OPR) and cancellation rate. Results: Reproductive outcomes were much better in the modified GnRH antagonist protocol. The OPR and LBR were much higher in the GnRH antagonist protocol group than in the GnRH agonist long protocol group [odds ratio (OR) 3.82, 95% confidence interval (CI) 1.47, 10.61, P=0.018; OR 4.33, 95% CI 1.38, 13.60, P=0.008; respectively]. Meanwhile, the cancellation rate was much lower in the GnRH antagonist protocol group (OR 0.13, 95% CI 0.02, 0.72; P=0.014). Mean LH level during stimulation did not have a predictive value on live birth. However, it was independently associated with the occurrence of ongoing pregnancy (OR 2.70, 95% CI 1.25, 5.85; P=0.01). The results of sensitivity analyses were consistent with the data mentioned above. The patients got completely different and excellent clinical outcomes in their second cycles stimulated with the modified GnRH antagonist protocol. Conclusion: Patients with deep ovarian suppression following GnRH agonist long protocol may benefit from a modified GnRH antagonist protocol based on LH levels.

Impacts of Cyclosporin A on clinical pregnancy outcomes of patients with a history of unexplained transfer failure: a retrospective cohort study.

BACKGROUND: A rapid development in assisted reproductive technology (ART) has led to a surge in its popularity among target couples. However, elucidation on the molecular mechanism and effective solutions for a common problem posed by ART, namely transfer failure, is still lacking. The new therapeutic potential of cyclosporin A (CsA), a typical immunosuppressant widely used in the treatment of rejection after organ transplantation, in recurrent pregnancy loss (RPL) patients may inspire some novel transfer failure therapies in the future. To further explore the clinical effects of CsA, this study investigated whether its application can improve clinical pregnancy outcomes in patients with a history of unexplained transfer failure in frozen-thawed embryo transfer (FET) cycles. METHODS: Data from a retrospective cohort investigation (178 frozen-thawed embryo transfer cycles in 178 patients) were analysed using binary logistic regression to explore the relationship between CsA treatment and clinical pregnancy outcomes; the odds ratios (ORs) and 95 % confidence intervals (CIs) were calculated as a measure of relevancy. Implantation rate was the main outcome measure. RESULTS: There was no difference in the fine adjusted OR (95 % CI) of the implantation rate [1.251 (0.739-2.120)], clinical pregnancy rate [1.634 (0.772-3.458)], chemical pregnancy rate [1.402 (0.285-6.909)], take-home baby rate [0.872 (0.423-1.798)], multiple births rate [0.840 (0.197-3.590)], preterm birth [1.668 (0.377-7.373)], abnormal birth weight [1.834 (0.533-6.307)] or sex ratio [0.956 (0.339-2.698)] between the CsA-treated group and control group. No birth defects were observed in the present study. CONCLUSIONS: Although CsA does not affect infant characteristics, it has no beneficial effects on the clinical pregnancy outcomes in patients with a history of unexplained transfer failure in FET cycles.

Effect of luteinizing hormone concentration on transcriptome and subcellular organelle phenotype of ovarian granulosa cells.

RESEARCH QUESTION: Granulosa cells (GCs) surrounding oocytes are crucial for follicular growth, oocyte development, ovulation, and luteinization under the dynamic co-stimulation of follicle stimulating hormone (FSH) and luteinizing hormone (LH). This study aimed to investigate the effect of LH levels on GCs in preovulatory follicles under gonadotropin releasing hormone antagonist-based ovarian stimulation. In vitro experiments were also conducted to study the direct effect of LH on GCs. METHODS: Twelve infertile women were divided into low (L), medium (M), and high (H) LH groups according to their serum LH levels during ovarian stimulation. RNA-sequencing (RNA-seq) was conducted to examine the transcriptome profiles of GCs obtained from the above patients during the oocyte retrieval. The activity of mitochondrial dehydrogenase was measured under the stimulation of recombinant LH (rLH) concentration gradient combined with recombinant FSH. The ultrastructures of subcellular organelles were observed. RESULTS: Bioinformatic analyses showed that compared with the M group, molecule and pathway changes in the L group and in the H group were similar. In cultured GCs, both insufficient and excessive rLH impaired the activity of mitochondrial dehydrogenase. With the medium rLH concentration, numerous cell connections and abundant mitochondria and liposomes were observed. Compared with the medium concentration, GCs showed smaller and rounder mitochondria, more autophagosomes, and massive organelles damages with excessive rLH, and swollen, circular, or forked mitochondria were observed with inadequate rLH. CONCLUSIONS: RNA-seq provided a novel spectrum of transcriptome characteristics of GCs potentially affected by serum LH levels during ovarian stimulation. In vitro, rLH could directly affect GCs at the subcellular level.

Lappaconitine sulfate induces apoptosis and G0/G1 phase cell cycle arrest by PI3K/AKT signaling pathway in human non-small cell lung cancer A549 cells.

Lappaconitine sulfate (LS) has good solubility and bioavailability. We have previously studied the anti-proliferative activity of LS on colon cancer HT-29 cell, but its anti-proliferative activity and molecular mechanism on human non-small cell lung cancer A549 cells are still unclear. This study was to investigate the effects of LS on proliferation, cell cycle and apoptosis in human non-small cell lung cancer A549 cells, and its possible molecular mechanisms. Cell proliferation activity was measured by Cell Counting Kit-8 (CCK-8) and 5-Ethynyl-2'- deoxyuridine (EdU) cell proliferation kit. Cell cycle was detected by propidium iodide (PI) flow cytometry. Apoptosis was detected by Annexin-V-FITC/PI method. Western blot was used to detect cycle and apoptosis-related proteins expression. These results showed that the proliferation activity of LS was significantly decreased in A549 cells, showing a dose- and time-dependent manner (p < 0.05). LS could increase the proportion of G0/G1 phase cells and decrease the proportion of cells in S phase, showing obvious G0/G1 phase arrest. LS significantly inhibited the expression of p-PI3K/PI3K, p-AKT/AKT, Cyclin D1 and Bcl-2 proteins (p < 0.05), and increased the expression of p53, p21, Bax, caspase 3 and caspase 9 (p < 0.05). Moreover, PI3K inhibitor (LY294002) significantly decreased A549 cell viability rate induced by LS, abrogated the activation of p-PI3K/PI3K and p-AKT/AKT in the presence of LS. These results indicated that LS could block A549 cells in the G0/G1 phase, induce apoptosis, and inhibit cell proliferation through the PI3K/AKT signaling pathway.

Multiple-dose versus single-dose gonadotropin-releasing hormone agonist after first in vitro fertilization failure associated with luteal phase deficiency: A randomized controlled trial.

OBJECTIVE: To evaluate the efficacy and safety of multiple- versus single-dose gonadotropin-releasing hormone agonist (GnRH-a) addition to luteal phase support (LPS), in patients with a first in vitro fertilization (IVF) failure associated with luteal phase deficiency (LPD). METHODS: Eighty patients with a first IVF failure associated with LPD were randomly assigned into single-dose and multiple-dose GnRH-a groups. In the second IVF attempt, patients in the single-dose group were given standard LPS plus a single dose of GnRH-a 6 days after oocyte retrieval. Patients in the multiple-dose group received standard LPS plus 14 daily injections of GnRH-a. Children conceived were followed up for 2 years. RESULTS: Pregnancy (67.5% vs. 42.5%), clinical pregnancy (50.0% vs. 22.5%), and live birth rates (42.5% vs. 20.0%) were significantly higher in the multiple-dose versus single-dose GnRH-a group. Patients in the multiple-dose GnRH-a group had significantly higher progesterone levels 14 days after oocyte recovery (35.9 vs. 21.4 ng/mL). No significant difference existed in the status at birth or developmental and behavior assessments of 2-year-old children conceived in both groups. CONCLUSIONS: Daily addition of GnRH-a to standard LPS can achieve better pregnancy outcomes with a sustained safety profile in patients with a first IVF failure associated with LPD.

Involvement of CXCL12/CXCR4 in the motility of human first-trimester endometrial epithelial cells through an autocrine mechanism by activating PI3K/AKT signaling.

BACKGROUND: CXCL12(chemokine ligand 12, CXCL12) and its receptors CXCR4 are widely expressed in maternal-fetal interface and plays an adjust role in materno-fetal dialogue and immune tolerance during early pregnancy. This study aimed to evaluate the role and mechanism of self-derived CXCL12 in modulating the functions of human first-trimester endometrial epithelial cells (EECs) and to identify the potential protein kinase signaling pathways involved in the CXCL12/CXCR4's effect on EECs. METHODS: The expression of CXCL12 and CXCR4 in EECs was measured by using immunohistochemistry, immunofluorescence, real-time polymerase chain reaction and enzyme-linked immunosorbent assay. The effects of EEC-conditioned medium (EEC-CM) and recombinant human CXCL12 (rhCXCL12) on EEC migration and invasion in vitro were evaluated with migration and invasion assays. In-cell western blot analysis was used to examine the phosphorylation of protein kinase B (AKT), extracellular regulated protein kinases (ERKs) and phosphatidylinositol 3-kinase (PI3K) after CXCL12 treatment. RESULTS: CXCL12 and CXCR4 were both expressed in human first-trimester EECs at the mRNA and protein level. Both EEC-CM and rhCXCL12 significantly increased the migration and invasion of EECs (P < 0.05), which could be blocked by neutralizing antibodies against CXCR4 (P < 0.05) or CXCL12 (P < 0.05), respectively. CXCL12 activated both PI3K/AKT and ERK1/2 signaling and CXCR4 neutralizing antibody effectively reduced CXCL12-induced phosphorylation of AKT and ERK1/2. LY294002, a PI3K-AKT inhibitor, was able to reverse the promotive effect of EEC-CM or rhCXCL12 on EEC migration and invasion. CONCLUSIONS: Human first-trimester EECs promoted their own migration and invasion through the autocrine mechanism with CXCL12/CXCR4 axis involvement by activating PI3K/AKT signaling. This study contributes to a better understanding of the epithelium function mediated by chemokine and chemokine receptor during normal pregnancy.

Contribution of soybean polysaccharides in digestion of oil-in-water emulsion-based delivery system in an in vitro gastric environment.

This study aims to evaluate the effects of soy soluble polysaccharide and soy hull polysaccharide on stability and characteristics of emulsions stabilised by soy protein isolate in an in vitro gastric environment. Zeta potential and particle size were used to investigate the changes of physico-chemical and stability in the three emulsions during in vitro gastric digestion, following the order: soy protein isolate-stability emulsion < soy protein isolate-soy soluble polysaccharide -stability emulsion < soy protein isolate-soy hull polysaccharide-stability emulsion, confirming that coalescence in the soy protein isolate-stability emulsion occurred during in vitro gastric digestion. Optical microscopy and stability measurement (backscattering) also validate that addition of polysaccharide (soy soluble polysaccharide and soy hull polysaccharide) can reduce the effect of simulated gastric fluid (i.e., pH, ionic strength and pepsin) on emulsion stability, especially, soy protein isolate-soy hull polysaccharide-stability emulsion, compared with soy protein isolate-stability emulsion. This suggests that the flocculation behaviours of these emulsions in the stomach lead to a difference in the quantity of oil and the size and structure of the oil droplets, which play a significant role in emulsion digestion in the gastrointestinal tract. This work may indicate a potential application of soy hull polysaccharide for the construction of emulsion food delivery systems.

Vitamin D improves in-vitro fertilization outcomes in infertile women with polycystic ovary syndrome and insulin resistance.

BACKGROUND: The aim of f this study is to investigate the effect of vitamin D supplementation upon the outcome of in-vitro fertilization (IVF) in infertile women with polycystic ovarian syndrome (PCOS) and insulin resistance (IR). METHODS: Three hundred and five infertile patients with PCOS and insulin resistance undergoing IVF were included in this study. All participants underwent oral glucose tolerance test (OGTT). Insulin resistance was calculated by homeostasis model assessment. Vitamin D status was measured by assessing circulating levels of 25OH-VD in serum samples by radioimmunoassay. RESULTS: All the patients were then divided into four groups according to their relative levels of serum 25OH-VD (levels of 25OH-VD≥20 ng/mL were defined as being normal) and whether treatment had been administered prior to COH: Group I (deficiency group without treatment [25OH-VD<20 ng/mL]); Group II (normal group [25OH-VD≥20 ng/mL]), Group III (normal group after treatment), Group IV (deficiency group after treatment). In total, 305 women were included in this study. Implantation rates across the four groups were 8.5% (9/106) in Group I, 49% (24/49) in Group II, 49.1% (55/112) in Group III and 14.3 (18/126) in Group IV, respectively. Clinical pregnancy rates were 19.3% (11/57), 65.2 (15/23), 66.7% (38/57) and 23.5% (16/68) in the four groups, respectively. Both the implantation rate and clinical pregnancy rate in groups in which serum 25OH-VD was normal (Groups II and III) were significantly higher (P<0.05) than the other two groups. Serum levels of 25OH-VD were highly correlated with clinical pregnancy and implantation rates (P<0.01).There also were significant differences among the four groups in terms of fertilization rate (80±16%, 93±10%, 90±12%, and 79±23%), two pronucleus (2PN) fertilization rate (59±21%, 72±14%, 76±17% and 66±24%) and cleavage rate (76±20%, 91±10%, 89±24% and 76±25%). Particularly marked differences were observed in the number of high-quality embryos (20±16%, 63±25%, 55±22%, and 32±19%) and the available embryos which could be transferred (2.5±1.8, 5.8±2.8, 5.2±2.4, and 3.6±1.9) (all P<0.01). Much higher rates of implantation and clinical pregnancy were observed among the 25OH-VD deficient patients who had received vitamin D supplements and 25OH-VD levels returned to normal compared to patients in which 25OH-VD levels did not return to normal. Group III, which had normal 25OH-VD levels following Vitamin D supplementation produced the same number of high-quality embryos as Group II which had normal 25OH-VD levels (Group III versus Group II; 7.6±4.1 vs. 10.6±5.2, 55±22% vs. 63±25%; P<0.05). CONCLUSIONS: Our data indicate that vitamin D supplementation can help return serum vitamin D levels in infertile women with PCOS and IR to normal levels leading to an improvement in the quality of embryos and a significantly higher clinical pregnancy rate. Maintaining a normal serum vitamin D level in PCOS women is very important in achieving a successful clinical pregnancy following in-vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI).

The decline in serum estradiol on the second day after oocyte retrieval affects the outcome of IVF/ICSI-ET treatment in high ovarian responders.

OBJECTIVE: To investigate the predictive value of the decline in serum estradiol on the second day after oocyte retrieval on the outcomes of in vitro fertilization (IVF) or intra-cytoplasmic sperm injection and embryo transfer (ICSI-ET) among high ovarian responders. DESIGN: Retrospective single-center cohort study. SETTING: Tertiary-care, university-affiliated teaching hospital. Patients Women aged 20-45 years undergoing assisted reproduction treatment from June 2014 to December 2015. INTERVENTIONS: A total of 980 cycles were included; 395 high responders (Group 1) and 256 normal responders (Group 3) underwent embryo transfer (ET) in fresh ET cycles. A total of 329 high ovarian responders who underwent cryopreservation of all embryos (Group 2) were recruited as controls. The cases were divided into the following five subgroups according to the rate of serum estradiol decline on the second day after oocyte retrieval: 50.00-59.99% (Subgroup A), 60.00-69.99% (Subgroup B), 70.00-79.99% (Subgroup C), 80.00-89.99% (Subgroup D) and ≥ 90.00% (Subgroup E). The clinical outcomes were analyzed. MAIN OUTCOME MEASURES: Clinical pregnancy rate, implantation rate. RESULTS: In Group 1, the pregnancy rate decreased from 51.33 to 36.72% and the implantation rate decreased from 30.93 to 21.70% when the level of serum estradiol on the second day after oocyte retrieval decreased by more than 80%, which was a statistically significant decline (p < 0.05). The peak estradiol (E2) value and implantation rate were also significantly different (p < 0.05). In Group 2, the decline of serum E2 on the second day after oocyte retrieval had no significant effect on the clinical pregnancy rate or the implantation rate. The trend was similar in Group 3. CONCLUSIONS: A decline in the E2 level of > 80% after oocyte retrieval may play an important role in unsatisfactory IVF/ICSI-ET outcomes among high ovarian responders.

ZBTB7 overexpression contributes to malignancy in breast cancer.

ZBTB7 was recently recognized as a proto-oncogene. We studied its prognostic value and relationship to clinicopathological variables in 125 breast cancer patients. ZBTB7 expression was significantly higher in breast cancer tissues than in normal breast tissues, but its gene amplification copies were relatively low. ZBTB7 expression levels were significantly correlated with histological grade (p = .023) and marginally inversely correlated with the presence of estrogen receptors (p = .053); its overexpression significantly predicted shorter recurrence-free survival (p = .033). Our results showed that ZBTB7 might be implicated in breast cancer development and may serve as a promising prognostic marker.

Increased expression of Nlp, a potential oncogene in ovarian cancer, and its implication in carcinogenesis.

OBJECTIVE: Nlp (Ninein-like protein), a novel centrosome protein involved in microtubule nucleation, has been studied extensively in our laboratory, and its overexpression has been found in some human tumors. To understand the role of Nlp in human ovarian cancer development, we studied the correlation of Nlp expression with clinicopathological parameters and survival in epithelial ovarian cancer, and the impact of Nlp overexpression on ovarian cancer cells. METHODS: Nlp expression in normal, borderline, benign and malignant epithelial ovarian tissues was examined by immunohistochemistry. The correlation between Nlp expression and tumor grade, FIGO stage and histological type was also evaluated. Survival was calculated using Kaplan-Meier estimates. Cell proliferation and apoptosis were assayed after stable transfection of pEGFP-C3-Nlp or empty vector in human ovarian cancer cell line SKOV3. RESULTS: Nlp was positive in 1 of 10 (10%) normal ovarian tissues, 5 of 34 (14.7%) benign tumors, 9 of 26 (34.6%) borderline tumors and 73 of 131 (56.0%) ovarian tumors. Nlp immunoreactivity intensity significantly correlated with tumor grade, but not with FIGO stage or histological type. Kaplan-Meier curves showed that Nlp overexpression was marginally associated with decreased overall survival. Overexpression of Nlp enhanced proliferation and inhibited apoptosis induced by paclitaxel in the SKOV3 cell line. CONCLUSIONS: Overexpression of Nlp in ovarian tumors raises the possibility that Nlp may play a role in ovarian carcinogenesis.

Telomerase expression and cell proliferation in ovarian cancer cells induced by histone deacetylase inhibitors.

OBJECTIVE: Telomerase is composed primarily of catalytic subunit (hTERT) and RNA template (hTERC). Histone deacetylase (HDAC) inhibitors are known to modulate transcription and change the expression of hTERT and hTERC mRNA and telomerase activity in several types of cancer cells, but it is unclear if there is a similar effect in ovarian cancer cells. METHOD: The present study was designed to evaluate the effects of HDAC inhibitors on hTERT and hTERC mRNA expression in ovarian cancer cells. SK-V-3 cells were treated with the HDAC inhibitors, trichostatinA (TSA) and sodium butyrate (NaB); the expression of hTERC and hTERT mRNA and telomerase activity were evaluated by RT-PCR and TRAP assay, respectively. RESULTS: In SK-OV-3 cells, TSA and NaB inhibited cell proliferation and induced apoptosis. The expression of hTERT and hTERT mRNA was not suppressed even after treatment with 1.0 microM TSA and 6 mM NaB, respectively. The telomerase activity was not changed by either TSA or NaB. CONCLUSION: Histone deacetylase inhibitors inhibited cell proliferation and induced apoptosis, but had no effect on the expression of hTERC and hTERT mRNA and on telomerase activity.