Coumestrol induces mitochondrial dysfunction by stimulating ROS production and calcium ion influx into mitochondria in human placental choriocarcinoma cells.

STUDY QUESTION: Does coumestrol inhibit proliferation of human placental choriocarcinoma cells? SUMMARY ANSWER: Coumestrol promotes cell death in the choriocarcinoma cells by regulating ERK1/2 MAPK and JNK MAPK signaling pathways and through disruption of Ca2+ and ROS homeostasis. WHAT IS KNOWN ALREADY: A number of patients who suffer from choriocarcinomas fail to survive due to delayed diagnosis or a recurrent tumor and resistance to traditional chemotherapy using platinum-based agents and methotrexate. To overcome these limitations, it is important to discover novel compounds which have no adverse effects yet can inhibit the expression of a target molecule to develop, as a novel therapeutic for prevention and/or treatment of choriocarcinomas. STUDY DESIGN, SIZE, DURATION: Effects of coumestrol on human placental choriocarcinoma cell lines, JAR and JEG3, were assessed in diverse assays in a dose- and time-dependent manner. PARTICIPCANTS/MATERIALS, SETTING, METHODS: Effects of coumestrol on cell proliferation, apoptosis (annexin V expression, propidium iodide staining, TUNEL and invasion assays), mitochondria-mediated apoptosis, production of reactive oxygen species (ROS), lipid peroxidation, glutathione levels and endoplasmic reticulum (ER) stress proteins in JAR and JEG3 cells were determined. Signal transduction pathways in JAR and JEG3 cells in response to coumestrol were determined by western blot analyses. MAIN RESULTS AND THE ROLE OF CHANCE: Results of the present study indicated that coumestrol suppressed proliferation and increased apoptosis in JAR and JEG3 cells by inducing pro-apoptotic proteins, Bax and Bak. In addition, coumestrol increased ROS production, as well as lipid peroxidation and glutathione levels in JAR and JEG3 cells. Moreover, coumestrol-induced depolarization of mitochondrial membrane potential (MMP) and increased cytosolic and mitochondrial Ca2+ levels in JAR and JEG3 cells. Consistent with those results, treatment of JAR and JEG3 cells with a Ca2+ chelator and an inhibitor of IP3 receptor decreased coumestrol-induced depolarization of MMP and increased proliferation in JAR and JEG3 cells. LARGE SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: A lack of in vivo animal studies is a major limitation of this research. The effectiveness of coumestrol to induce apoptosis of human placental choriocarcinoma cells requires further investigation. WIDER IMPLICATIONS OF THE FINDINGS: Our results indicate that coumestrol induces apoptotic effects on placental choriocarcinoma cells by regulating cell signaling and mitochondrial-mediated functions, with a potential to impair progression of the cancer. STUDY FUNDING/COMPETING INTEREST(S): This research was supported by grants from the Korea Health Technology R&D Project through the Korea Health Industry Development Institute (KHIDI), funded by the Ministry of Health & Welfare, Republic of Korea (No. HI15C0810 awarded to G.S. and HI17C0929 awarded to W.L.).

Alpha Lipoic Acid (ALA) effects on subchorionic hematoma: preliminary clinical results.

OBJECTIVE: The clinic use of alpha Lipoic Acid (ALA) is linked to its capability to exert antioxidant effects and, more interestingly, to counteract the pathologic changes of complex networks of cytokines, chemokines and growth factors, restoring their physiological state. The aim of this randomized controlled clinical trial was to test the contribution of oral supplementation of ALA to the standard treatment with Progesterone vaginal suppositories, in healing subchorionic hematomas in patients with threatened miscarriage. Controls were administered only Progesterone suppositories. PATIENTS AND METHODS: Nineteen pregnant women in the first trimester of gestation, with threatened miscarriage and ultrasound evidence of subchorionic hematoma, were included in the trial and randomly divided in two groups: controls, treated with 400 mg Progesterone (200 mg 2 times per day), given by vaginal suppositories, and case study treated with the same Progesterone dosage, plus ALA, given orally at the dose of 600 mg (300 mg 2 times per day, DAV®, Lo.Li. Pharma srl, Italy). Sixteen patients completed the trial. Treatment was performed until complete resolution of the clinical picture. RESULTS: In both groups, the subjects improved significantly but, in general, a better and faster evolution in the major signs of threatened miscarriage was observed in the subjects treated with ALA and Progesterone. In these patients, the speed of resorption of subchorionic hematoma was significantly (p ≤ 0.05) superior compared to controls. The ALA and Progesterone group showed a faster decrease or disappearance of all symptoms than that observed in the control group, however the difference was not significant. CONCLUSIONS: These preliminary results suggest that ALA supplementation significantly contributes to speed up the process of restoration of physiological conditions in threatened miscarriage and ameliorates the medical conditions of both the mothers and the foetus, probably modulating the networks of cytokines, growth factors and other molecules.

Trombohematoma subcorionico masivo: una patología placentaria emergente / Massive subchorionic thrombohematoma

El trombohematoma subcoriónico es una extravasación de sangre localizada en la placa coriónica, entre amnios y corion. Es muy infrecuente, el diagnóstico no es común, tiene alto riesgo perinatal y no hay casos comunicados en nuestro medio. Se presentan 12 casos de sospecha diagnóstica antenatal, confirmada en el examen histopatológico placentario. Se describe y discute el cuadro clínico, las complicaciones maternas y perinatales, el diagnóstico ultrasónico, el manejo y los resultados obtenidos. En nueve casos se identifcó una fase latente con hematoma de tamaño estable, entre el inicio de los síntomas y el parto, que duró en promedio 7,3 semanas. En ocho casos la fase latente fue seguida por una fase activa con aumento del hematoma asociado al parto prematuro. Tres embarazadas presentaron patología médica compleja con una muerte materna. Seis casos hicieron anemia severa y tres patología miscelánea. Hubo ocho amenazas de parto prematuro con tocolisis, tres rotura prematura de membranas, una colestasis y una preeclampsia. Los partos fueron prematuros, dos de 36 y 33 semanas y diez menores a 32 semanas. Siete prematuros tuvieron peso inferior a 1000 gramos y seis hicieron restricción fetal grave, en percentil <5 de la curva de crecimiento. Hubo complicaciones neonatales relacionadas con prematurez, restricción y bajo peso, manejados con hospitalización prolongada con promedio de 74 días (rango: 6-298 días). Diez neonatos sobrevivieron; hubo un mortinato y un mortineonato. La sobrevida fue 83,3 por ciento y la mortalidad de 16,6 por ciento que se comparan favorablemente con las cifras comunicadas.

Subchorial thrombohaematoma is caused by blood extravasations in the corionic plate, between amnion and chorion. It is a rare pathologic entity, that carries a high perinatal risk, which has not being published in our country up to now. We report 12 cases in which the diagnosis was suspected before birth, and confirmed in the placentary pathological examination. We describe the clinical presentation, fetal and maternal risks, ultrasonographic findings, treatment and clinical outcomes. In 9 patients a latent phase was identified with a stable size hematoma, which had a mean duration of 7.3 weeks. In 8 cases the latent phase was followed by an active phase, with increasing size of the hematoma associated with preterm labour. Three pregnant women had severe complications which caused one maternal death. Six had severe anemia and other three had minor complications. Eight had preterm labor symptoms which required tocolysis. Three had prelabour rupture of membranes, one cholestasis disease and preeclampsia. Preterm labours were at 36, 33 and other ten before 32 weeks of gestation. Seven preterm newborns weight less than 1000 grams and six had severe fetal restriction (p<5). Newborn complications were related with prematurity, requiring prolonged hospitalization (mean 74 days, range 6-298 days). Ten newborns survived. There were 1 still birth and 1 dead newborn. Survival rate was 83.3 percent and 16.6 percent mortality, better rates than previously published.