cyclin-d1 and Choriocarcinoma

In this study, the effects of cigarette smoke (CS) on the induction of apoptosis via reactive oxygen species (ROS) production and endoplasmic reticulum stress (ER stress) of JEG-3 human choriocarcinoma cells were examined to confirm the relationship between CS and placenta development. Upon TUNEL assay, CS extract (3R4F; 0.3 and 2.1 μM) increased JEG-3 apoptosis. Western blot assay revealed that the protein expressions of p53, Bax, and CCAAT-enhancer-binding protein homologous protein (CHOP) increased, while the levels of Bcl-2 were reduced following CS extract treatment. Moreover, 2',7'-dichlorofluorescein diacetate (DCFH-DA) assay revealed increased ROS production. Upon 3-(4-5-dimethylthiazol-2-yl)-2.5-dyhphenyltetrazolium bromide (MTT) assay, isoprene (IP), one of ingredients of CS, deceased JEG-3 cell viability (10

Topics: Apoptosis; Autophagy; bcl-2-Associated X Protein; Butadienes; Cell Line, Tumor; Choriocarcinoma; Cyclin D1; Cyclin E; Endoplasmic Reticulum Stress; Female; Hemiterpenes; Humans; Kelch-Like ECH-Associated Protein 1; NF-E2-Related Factor 2; Nicotiana; Oncogene Proteins; Pentanes; Pregnancy; Reactive Oxygen Species; Sequestosome-1 Protein; Smoke; Transcription Factor CHOP; Tumor Suppressor Protein p53; Uterine Neoplasms

Cigarette smoke (CS) causes about 480,000 deaths each year worldwide, and it is well-known to have harmful effects on the human body, leading to heart disease, stroke, lung cancer, and cardiovascular problems. In this study, the effects of formaldehyde (FA) and benzene (Bz), the main components of CS, on cell proliferation and epithelial mesenchymal transition (EMT) of JEG-3 human choriocarcinoma cells were examined to confirm the relationship between CS components and placenta carcinoma. Upon MTT assay, FA (10

Topics: Antigens, CD; Antioxidants; Benzene; Cadherins; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Choriocarcinoma; Cyclin D1; Epithelial-Mesenchymal Transition; Female; Formaldehyde; Gene Expression Regulation; Humans; Pregnancy; Reactive Oxygen Species

KN-93, a membrane-permeant calcium/calmodulin- dependent kinase-selective inhibitor, induces apoptosis in some lines of human tumor cells. We investigated the effect of KN-93 in the choriocarcinoma cell line, BeWo. BeWo cells were treated with various concentrations of KN-93, and changes in cell growth, the cell cycle, apoptosis, and related parameters were examined. A WST-1 assay showed that BeWo cells were sensitive to the growth inhibitory effect of KN-93. Cell cycle analysis indicated that exposure to KN-93 decreased the proportion of cells in the S phase and increased the proportion in the G0/G1 phases of the cell cycle. Induction of apoptosis was confirmed by Annexin V staining of externalized phosphatidylserine, by the loss of mitochondrial transmembrane potential, and by antibodies directed against histones from fragmented DNA. This induction occurred in conjunction with the altered expression of genes related to cell growth, malignant phenotype, and apoptosis. These results suggest that KN-93 may serve as a therapeutic agent for the treatment of choriocarcinoma.

Topics: Apoptosis; Benzylamines; Blotting, Western; Calcium-Calmodulin-Dependent Protein Kinases; Caspase 3; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cell Survival; Choriocarcinoma; Cyclin A; Cyclin D1; Dose-Response Relationship, Drug; Flow Cytometry; G1 Phase; Humans; Membrane Potential, Mitochondrial; Protein Kinase Inhibitors; Proto-Oncogene Proteins c-bcl-2; Resting Phase, Cell Cycle; S Phase; Sulfonamides

Leptin, the 16-kDa protein product of the obese gene, was originally considered as an adipocyte-derived signaling molecule for the central control of metabolism. However, leptin has been suggested to be involved in other functions during pregnancy, particularly in placenta. In the present work, we studied a possible effect of leptin on trophoblastic cell proliferation, survival, and apoptosis. Recombinant human leptin added to JEG-3 and BeWo choriocarcinoma cell lines showed a stimulatory effect on cell proliferation up to 3 and 2.4 times, respectively, measured by (3)H-thymidine incorporation and cell counting. These effects were time and dose dependent. Maximal effect was achieved at 250 ng leptin/ml for JEG-3 cells and 50 ng leptin/ml for BeWo cells. Moreover, by inhibiting endogenous leptin expression with 2 microM of an antisense oligonucleotide (AS), cell proliferation was diminished. We analyzed cell population distribution during the different stages of cell cycle by fluorescence-activated cell sorting, and we found that leptin treatment displaced the cells towards a G2/M phase. We also found that leptin upregulated cyclin D1 expression, one of the key cell cycle-signaling proteins. Since proliferation and death processes are intimately related, the effect of leptin on cell apoptosis was investigated. Treatment with 2 microM leptin AS increased the number of apoptotic cells 60 times, as assessed by annexin V-fluorescein isothiocyanate/propidium iodide staining, and the caspase-3 activity was increased more than 2 fold. This effect was prevented by the addition of 100 ng leptin/ml. In conclusion, we provide evidence that suggests that leptin is a trophic and mitogenic factor for trophoblastic cells by virtue of its inhibiting apoptosis and promoting proliferation.

Topics: Apoptosis; Caspase 3; Cell Cycle; Cell Division; Cell Line, Tumor; Cell Proliferation; Cell Survival; Choriocarcinoma; Cyclin D1; Dose-Response Relationship, Drug; Female; Flow Cytometry; G2 Phase; Humans; Leptin; Pregnancy; Protein Isoforms; Receptors, Leptin; Recombinant Proteins; Time Factors; Trophoblasts; Up-Regulation; Uterine Neoplasms