ABSTRACT
Melanoma, an aggressive skin tumor with high metastatic potential, is associated with high mortality and increasing morbidity. Multiple available chemotherapeutic and immunotherapeutic modalities failed to improve survival in advanced disease, and the search for new agents is ongoing. The aim of this study was to investigate antimelanoma effects of O,O-diethyl-(S,S)-ethylenediamine-N,N'di-2-(3-cyclohexyl) propanoate dihydrochloride (EE), a previously synthesized and characterized organic compound. Mouse melanoma B16 cell viability was assessed using acid phosphatase, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, sulforhodamine B, and lactate dehydrogenase assays. Apoptosis and autophagy were investigated using flow cytometry, fluorescence and electron microscopy, and western blotting. In vivo antitumor potential was assessed in subcutaneous mouse melanoma model after 14 days of treatment with EE. Tumor mass and volume were measured, and RT-PCR was used for investigating the expression of autophagy-related, proapoptotic, and antiapoptotic molecules in tumor tissue. Investigated organic compound exerts significant cytotoxic effect against B16 cells. EE induced apoptosis, as confirmed by phosphatidyl serine externalisation, caspase activation, and ultrastructural features typical for apoptosis seen on fluorescence and electron microscopes. The apoptotic mechanism included prompt disruption of mitochondrial membrane potential and oxidative stress. No autophagy was observed. Antimelanoma action and apoptosis induction were confirmed in vivo, as EE decreased mass and volume of tumors, and increased expression of several proapoptotic genes. EE possesses significant antimelanoma action and causes caspase-dependent apoptosis mediated by mitochondrial damage and reactive oxygen species production. Decrease in tumor growth and increase in expression of proapoptotic genes in tumor tissue suggest that EE warrants further investigation as a candidate agent in treating melanoma.
Subject(s)
Antineoplastic Agents/pharmacology , Apoptosis/drug effects , Cyclohexanes/pharmacology , Ethylenediamines/pharmacology , Melanoma, Experimental/pathology , Membrane Potential, Mitochondrial/drug effects , Oxidative Stress/drug effects , Propionates/pharmacology , Animals , Autophagy , In Vitro Techniques , Melanoma, Experimental/drug therapy , Mice , Mice, Inbred C57BL , Reactive Oxygen Species/metabolism , Tumor Cells, CulturedABSTRACT
STUDY OBJECTIVE: Exploring the relation between the age, time since menarche, anthropometric parameters and the growth of the uterus and ovaries in postmenarcheal girls. DESIGN: Cross sectional. SETTING: Department of Human reproduction at a tertiary pediatric referral center. PARTICIPANTS: Eight hundred thirty-five adolescent girls. INTERVENTIONS: Postmenarcheal girls were classified according to the regularity of their menstrual cycles in 2 groups (regular and irregular cycles) and compared. Anthropometric measurements and ultrasonographic examination of the pelvis was conducted with all participants. MAIN OUTCOME MEASURES: Anthropometric and ultrasonographic parameters were evaluated. RESULTS: Results of our study showed that girls with regular and irregular cycles differed in height, weight, body mass index, percentage of body fat and ovarian volumes. The size of the ovaries decreases in the group of girls with regular cycles (r = 0.14; P < .005), while it increases in girls with irregular cycles (r = 0.15; P < .001) with advancing age. Uterine volume in all patients increases gradually with age reaching consistent values at 16 years (r = 0.5; P < .001). Age at menarche, the time elapsed since menarche, the height, weight, body mass index and percentage of body fat in patients correlated with uterine volume. Ovarian volume correlated with patients' weight, BMI and percentage of fat. CONCLUSION: Uterus continues to grow in postmenarcheal years, with increasing height and weight of girls, regardless of the regularity of cycles. Postmenarcheal girls with irregular cycles were found to have heavier figures and larger ovaries.
Subject(s)
Body Composition , Menstrual Cycle/physiology , Menstruation Disturbances/physiopathology , Ovary/growth & development , Uterus/growth & development , Adiposity , Adolescent , Age Factors , Body Height , Body Mass Index , Body Weight , Child , Cross-Sectional Studies , Female , Humans , Menarche/physiology , Organ Size , Ovary/diagnostic imaging , Ultrasonography , Uterus/diagnostic imagingABSTRACT
The excellent photoluminescent properties of graphene quantum dots (GQD) makes them suitable candidates for biomedical applications, but their cytotoxicity has not been extensively studied. Here we show that electrochemically produced GQD irradiated with blue light (470 nm, 1W) generate reactive oxygen species, including singlet oxygen, and kill U251 human glioma cells by causing oxidative stress. The cell death induced by photoexcited GQD displayed morphological and/or biochemical characteristics of both apoptosis (phosphatidylserine externalization, caspase activation, DNA fragmentation) and autophagy (formation of autophagic vesicles, LC3-I/LC3-II conversion, degradation of autophagic target p62). Moreover, a genetic inactivation of autophagy-essential LC3B protein partly abrogated the photodynamic cytotoxicity of GQD. These data indicate potential usefulness of GQD in photodynamic therapy, but also raise concerns about their possible toxicity.
