Wnt/ß-Catenin Inhibition by CWP232291 as a Novel Therapeutic Strategy in Ovarian Cancer.

The poor prognosis of ovarian cancer patients mainly results from a lack of early diagnosis approaches and a high rate of relapse. Only a very modest improvement has been made in ovarian cancer patient survival with traditional treatments. More targeted therapies precisely matching each patient are strongly needed. The aberrant activation of Wnt/ß-catenin signaling pathway plays a fundamental role in cancer development and progression in various types of cancer including ovarian cancer. Recent insight into this pathway has revealed the potential of targeting Wnt/ß-catenin in ovarian cancer treatment. This study aims to investigate the effect of CWP232291, a small molecular Wnt/ß-catenin inhibitor on ovarian cancer progression. Various in vitro, in vivo and ex vivo models are established for CWP232291 testing. Results show that CWP232291 could significantly attenuate ovarian cancer growth through inhibition of ß-catenin. Noticeably, CWP232291 could also s suppress the growth of cisplatin-resistant cell lines and ovarian cancer patient-derived organoids. Overall, this study has firstly demonstrated the anti-tumor effect of CWP232291 in ovarian cancer and proposed Wnt/ß-catenin pathway inhibition as a novel therapeutic strategy against ovarian cancer.

Suppression of gain-of-function mutant p53 with metabolic inhibitors reduces tumor growth in vivo.

Mutation of p53 occasionally results in a gain of function, which promotes tumor growth. We asked whether destabilizing the gain-of-function protein would kill tumor cells. Downregulation of the gene reduced cell proliferation in p53-mutant cells, but not in p53-null cells, indicating that the former depended on the mutant protein for survival. Moreover, phenformin and 2-deoxyglucose suppressed cell growth and simultaneously destabilized mutant p53. The AMPK pathway, MAPK pathway, chaperone proteins and ubiquitination all contributed to this process. Interestingly, phenformin and 2-deoxyglucose also reduced tumor growth in syngeneic mice harboring the p53 mutation. Thus, destabilizing mutant p53 protein in order to kill cells exhibiting "oncogene addiction" could be a promising strategy for combatting p53 mutant tumors.

Soybean-derived glyceollins induce apoptosis through ROS generation.

Glyceollins, which are synthesized from daidzein in soybeans infected with fungi, have been shown to have anti-fungal effects and antioxidant properties. However, the anti-proliferative mechanism of glyceollins against tumor cells is unknown. Glyceollin-induced apoptosis was evidenced by a decrease in cell viability and mitochondrial membrane potential, and an increase in early redistribution of plasma membrane phosphatidylserine, the sub G1 phase, and DNA fragmentation in hepa1c1c7 cells. Western blot analysis showed that treatment of the hepa1c1c7 cells with the glyceollins decreased the expression of pro-caspase-3, Bcl-2, and cell cycle-related proteins, but increased the expression of p21 and p27, and cytochrome C release into cytosol. At a concentration of 6 µg mL(-1) or higher, glyceollins significantly stimulated the production of reactive oxygen species (ROS), which appear to be responsible for the apoptotic activity of the compounds. Our present study demonstrated that the high dose of glyceollins possibly caused apoptosis in mouse hepatoma cells through the production of ROS, suggesting the potential to exploit glyceollins as anti-tumorigenic agents.

A KCNQ1 mutation causes age-dependant bradycardia and persistent atrial fibrillation.

Atrial fibrillation (AF) is the most common arrhythmia. Gain-of-function mutations in KCNQ1, the pore-forming α-subunit of the slow delayed rectifier K current (IKs) channel, have been associated with AF. The purpose of this study was functional assessment of a mutation in KCNQ1 identified in a family with persistent AF and sinus bradycardia. We investigated whether this KCNQ1 missense mutation could form the genetic basis for AF and bradycardia simultaneously in this family. Sanger sequencing in a family with hereditary persistent AF identified a novel KCNQ1 variant (V241F) in a highly conserved region of S4 domain. The proband and her son developed bradycardia and persistent AF in an age-dependent fashion. The other son was a mutation carrier but he showed sinus bradycardia and not AF. Whole-cell patch clamp electrophysiology showed that V241F mutation in KCNQ1 shifted the activation curve to the left and dramatically slowed deactivation, leading to a constitutively open-like phenotype. Computer modeling showed that V241F would slow pacemaker activity. Also, simulations of atrial excitation predicted that V241F results in extreme shortening of action potential duration, possibly resulting in AF. Our study indicates that V241F might cause sinus bradycardia by increasing IKs. Additionally, V241F likely shortens atrial refractoriness to promote a substrate for reentry. KCNQ1 mutations have previously been described in AF, yet this is the first time a mutation in KCNQ1 is associated with age-dependent bradycardia and persistent AF. This finding further supports the hypothesis that sinus node dysfunction contributes to the development of AF.

A case of IgM deficiency with B cell deficiency detected by ABO discrepancy in a patient with acute osteomyelitis.

ABO discrepancy refers to an inconsistency between red cell and serum typings and has various causes, including hypogammaglobulinemia. IgM deficiency is a rare disorder that may accompany several conditions such as infection and autoimmune disorders. Here, we describe a case of IgM deficiency discovered during the evaluation of an ABO discrepancy in a 16-yr-old Korean boy. ABO blood grouping showed that while his cell type was O+, serum typing detected only anti-A (3+). Anti-B was not detectable at room temperature but was graded at 1+ at 4℃. ABO genotyping revealed an O/O genotype. His serum IgG, IgA, and IgM concentrations were 770 mg/dL (reference range: 800-1,700 mg/dL), 244 mg/dL (reference range: 100-490 mg/dL), and 13.5 mg/dL (reference range: 50-320 mg/dL), respectively. He was diagnosed with acute osteomyelitis on the basis of clinical presentation and imaging studies. The symptoms gradually improved within 3 weeks of treatment. However, the ABO discrepancy and IgM deficiency persisted even 6 months after recovery and lymphocyte subset analysis revealed CD19+ B cell deficiency. To the best of our knowledge, IgM deficiency detected by ABO discrepancy in a patient with acute osteomyelitis has not been reported before.

Synergistic activation of the Nrf2-signaling pathway by glyceollins under oxidative stress induced by glutathione depletion.

Oxidative stress state such as depletion of the intracellular glutathione (GSH) is associated with the development of cancer. Some dietary phytochemicals have been shown to possess a cancer preventive effect, although the understanding of the involved mechanisms is still limited. Recent study has shown that glyceollins, phytoalexins derived from soybean by biotic elicitor, might have a cancer preventive effect through induction of detoxifying/antioxidant enzymes. The objective of this study was to investigate the effects of glyceollins on the Nrf2 signaling pathway under excessive oxidative stress induced by GSH depletion. In mouse hepatoma cells (Hepa1c1c7) subjected to the buthionine sulfoximine (BSO), an inhibitor of γ-glutamylcysteine synthetase (γGCS), the intracellular GSH content was significantly lowered. On the other hand, incubation with glyceollins in the presence of BSO increased the level of GSH, expression of γGCS, and nuclear translocation of NF-E2-related factor-2 (Nrf2), compared to the cells treated with BSO only. Nrf2-antioxidant responsive element (ARE)-reporter activity assay in HepG2-C8 showed that BSO increased the ARE-reporter activity in a dose-dependent manner, compared to vehicle-treated cells, whereas cotreatment with glyceollins caused further increase in reporter luciferase activity relative to BSO alone. Taken together, glyceollins synergistically activated the Nrf2 signaling pathway and subsequently the expression of phase 2/antioxidant enzymes in the presence of BSO, suggesting that BSO-induced oxidative stress and that glyceollins regulate the expression of phase 2/antioxidant enzymes through different mechanisms from each other.

Atypical hereditary sensory and autonomic neuropathy type IV with neither mental retardation nor pain insensitivity.

Hereditary sensory and autonomic neuropathy type IV is an autosomal recessive disorder characterized by severe mental retardation and self-mutilation-related complications. Recently, we investigated a 16-year-old Korean boy with normal intelligence. He had preserved pain sensation but was suspected of having hereditary sensory and autonomic neuropathy type IV because of the recurrent bone fractures and painless joint destruction in the absence of any predisposing medical conditions. Genetic analysis of the NTRK1 gene revealed compound heterozygous mutations including c.851-33T>A and c.2303C>T (p.Pro768Leu) in the NTRK1 gene. The p.Pro768Leu mutation has been identified in 2 Japanese patients with a mild phenotype. Therefore, although it is rare, hereditary sensory and autonomic neuropathy type IV should be considered in patients with recurrent bone fractures and painless joint destruction who do not have any predisposing conditions even when they do not have typical clinical features such as mental retardation or pain insensitivity.

CKIT mutation in therapy-related acute myeloid leukemia with MLLT3/MLL chimeric transcript from t(9;11)(p22;q23).

Gain-of-function mutations of the CKIT gene have been reported to specifically occur in core-binding factor (CBF) acute myeloid leukemia (AML) with a poor prognostic implication. Here we report a case of therapy-related AML with t(9;11)(p22;q23) who had CKIT mutation. A 48-year-old woman with breast cancer received partial mastectomy followed by 6 cycles of adjuvant chemotherapy and radiation therapy. At 28 months from the diagnosis of breast cancer, she was diagnosed as having AML with blasts 81% in bone marrow. Cytogenetic analysis revealed t(9;11)(p22;q23), and FISH showed 96.5% of MLL break-apart signals. RT-PCR study revealed MLL(11q23)/MLLT3(9p22) chimeric transcript. FLT3-ITD and NPM1 mutations were both negative. Unexpectedly, mutation analyses for CKIT identified D816Y mutation. The patient received induction chemotherapy and achieved complete remission at 1 month. To the best of our knowledge, this is the first report on CKIT mutation in therapy-related AML with MLL rearrangement.

Hemoglobin Yamagata: hemoglobin variant detected by HbA1c test.

Hemoglobin (Hb) Yamagata is a rare Hb variant, which has been reported only twice-one case each in Japan and Korea. This variant arises from a Lys --> Asn substitution due to a mutation of AAA to AAC or AAT at codon 133 of the beta-globin gene. This study reports the third case of a patient detected with Hb Yamagata [HBB: c.399A>T; p.Lys133Asn] and discusses the effect of this variant on HbA1c measurement. This variant was detected in a 70-yr-old Korean man with diabetes mellitus during a routine follow-up. The HbA1c concentration determined using Variant ll Turbo (Bio-Rad, USA) was abnormally high at 47.9%. It was impossible to measure the HbA1c level accurately using Variant ll Thalassemia Mode (Bio-Rad, USA). However, the HbA1c levels analyzed by HLC-723 G7 (Tosoh, Japan), Cobas Integra (Roche, Switzerland) and NycoCard (Axis-Shield, Norway) were 5.0%, 8.0%, and 7.9%, respectively. This study shows that Hb Yamagata interferes with the accurate measurement of HbA1c levels in a diabetic patient. Taking these findings into consideration, we think that an immunoassay or affinity chromatography can be used as an alternate method for measuring the HbA1c level in a patient with this variant. In conclusion, a patient can be inferred to have an Hb variant if the HbA1c concentration is abnormally high or low or if there is a discrepancy between the results obtained using different methods, and if the clinical status of the patient suggests the presence of abnormal Hb. Subsequently, the HbA1c values can be determined by methods based on different principles.

[Clinical significance of minor elevation of cardiac troponin I].

BACKGROUND: Cardiac troponin I (cTnI) is known as a sensitive and specific marker for myocardial ischemia. The purposes of this study are to establish cut-off values of cTnI for acute myocardial infarction (AMI) and to analyze clinical significance of minor elevation of cTnI. METHODS: Two hundred and four patients from whom cTnI was measured at Ewha Womans University Dongdaemun hospital from January to March, 2006 were enrolled in the study. cTnI was measured using Dimension RxL (Dade Behring, USA). The lower limit of detection (LLD), 10% CV value, 99th percentile of healthy individuals, and cut-off value for AMI by ROC curve analysis were determined. RESULTS: LLD, 10% CV value, and 99th percentile of cTnI were 0.00 ng/mL, 0.10 ng/mL, and 0.07 ng/mL, respectively. The cut-off value of peak cTnI for AMI by ROC curve analysis was 0.13 ng/mL with the sensitivity, specificity, and AUC of 90.9%, 87.7%, and 0.921, respectively. The peak value of cTnI of patients with ischemic heart disease (IHD) was higher than that of the patients without IHD (P<0.05). According to the above reference and cut-off values of the initial cTnI, patients were categorized into four groups; < or =0.05 ng/mL (group 1), 0.06-0.09 ng/mL (group 2), 0.10-0.59 ng/mL (group 3), > or =0.60 ng/mL (group 4), and compared frequencies of AMI, IHD, cardio vascular disease (CVD) and death after 1 month among groups. Frequencies of AMI, IHD, CVD, and death after 1 month were significantly increased as the cTnI concentrations were increased (P<0.05). CONCLUSIONS: Minor elevation of cTnI value, even in group 3 was significantly associated with high incidence of AMI, IHD, CVD, and death rate after 1 month.