Mitochondrial dysfunction and transactivation of p53-dependent apoptotic genes in BaP-treated human fetal lung fibroblasts

Benzo(a)pyrene (BaP) has been shown to be an inducer of apoptosis. However, mechanisms involved in BaP-induced mitochondrial dysfunction are not well-known. In this study, human fetal lung fibroblasts cells were treated with BaP (8, 16, 32, 64 and 128 μM) for 4 and 12 h. Cell viability, intracellular level of reactive oxygen species (ROS), total antioxidant capacity (T-AOC), mitochondrial membrane potential (Δ Ψm) and cytochrome c release were determined. Changes in transcriptional levels of p53-dependent apoptotic genes ( p53, APAF1, CASPASE3, CASPASE9, NOXA and PUMA) were measured. At time point of 4 h, BaP induced the intracellular ROS generation in 64 ( p < .05) and 128 μM BaP groups ( p < .01) but decreased the T-AOC activities in 32, 64 ( p < .05 for both) and 128 μM BaP groups ( p < .01). At time point of 12 h, Δ Ψm significantly decreased in ≥32 μM BaP groups ( p < .05 for all). Amount of mitochondrial cytochrome c significantly increased in 128 μM BaP group ( p < .01). Transcriptional levels of CASPASE3, CASPASE9, APAF1 and PUMA were up-regulated in all BaP groups ( p < .05 for all) and in ≥32 μM groups for NOXA ( p < .05). But only in 16 μM BaP group a relatively little expression of p53 mRNA was observed ( p < .05). The results indicate that in the earlier period BaP promoted the generation of excessive ROS and subsequently the mitochondrial depolarization, whereas transactivations of the p53-dependent apoptotic genes were significantly induced at the later period.

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Collagen secretion mediated by NMDR activation in human fetal lung fibroblasts

Cell Biology International ◽

10.1042/cbi034s070a ◽

2010 ◽

Vol 34 (8) ◽

pp. S70-S70

Author(s):

MingJie WANG ◽

ZiQiang LUO ◽

Mei LU ◽

LiHong SHANG ◽

ShaoJie YUE

Keyword(s):

Fetal Lung ◽

Lung Fibroblasts ◽

Collagen Secretion ◽

Human Fetal Lung

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Calphostin C-mediated translocation and integration of Bax into mitochondria induces cytochrome c release before mitochondrial dysfunction

Cell Death and Differentiation ◽

10.1038/sj.cdd.4400682 ◽

2000 ◽

Vol 7 (6) ◽

pp. 511-520 ◽

Cited By ~ 25

Author(s):

H Ikemoto ◽

E Tani ◽

I Ozaki ◽

H Kitagawa ◽

N Arita

Keyword(s):

Cytochrome C ◽

Mitochondrial Dysfunction ◽

Cytochrome C Release ◽

Calphostin C

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MORPHOGENETIC ASPECTS OF MULTILAYERING IN PETRI DISH CULTURES OF HUMAN FETAL LUNG FIBROBLASTS

The Journal of Cell Biology ◽

10.1083/jcb.41.1.298 ◽

1969 ◽

Vol 41 (1) ◽

pp. 298-311 ◽

Cited By ~ 42

Author(s):

Tom Elsdale ◽

Robert Foley

Keyword(s):

Extracellular Matrix ◽

Human Lung ◽

Petri Dish ◽

Fetal Lung ◽

Single Species ◽

Lung Fibroblasts ◽

Human Lung Fibroblasts ◽

Spontaneous Aggregation ◽

Control Hypothesis ◽

Human Fetal Lung

Randomly seeded Petri dish cultures of embryonic human lung fibroblasts generate, in the course of their growth, highly ordered cellular arrangements. Thick, bilaterally symmetrical ridges with an axial polarity and an orthogonal, multilayered internal organization are observed within stationary cultures. The generation of these structures has been investigated. Ridges result from the spontaneous aggregation of cells in postconfluent cultures brought about by directed cell movements. These movements are promoted by the localized production of extracellular matrix sheets containing collagen, which provide new substrates for cellular colonization. Cells that have colonized one matrix substrate may secrete another above themselves, which will in turn be colonized. By a continuation of this cycle, thick stacks consisting of alternate layers of cells and matrix are produced to yield the observed aggregations. The distribution and shape of ridges in a culture imply that matrix substrates are confined to specific locations. The suggested control hypothesis assumes that all the cells in fibroblast cultures are potential producers of a single species of matrix. The serviceability of this matrix as a substrate for cellular colonization, however, is destroyed if the producer cells are motile. Matrix substrates, therefore, are only made by nonmotile cells.

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Prostacyclin analogs inhibit fibroblast migration

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.00432.2001 ◽

2002 ◽

Vol 283 (2) ◽

pp. L428-L432 ◽

Cited By ~ 25

Author(s):

Tadashi Kohyama ◽

Xiangde Liu ◽

Hui Jung Kim ◽

Tetsu Kobayashi ◽

Ronald F. Ertl ◽

...

Keyword(s):

Tissue Repair ◽

Fetal Lung ◽

Inhibitory Effect ◽

Lung Fibroblasts ◽

Fibroblast Migration ◽

Inflammatory Processes ◽

Dependent Protein ◽

Pka Pathway ◽

Human Fetal Lung ◽

Abnormal Tissue

The controlled accumulation of fibroblasts to sites of inflammation is crucial to effective tissue repair after injury. Either inadequate or excessive accumulation of fibroblasts could result in abnormal tissue function. Prostacyclin (PGI2) is a potent mediator in the coagulation and inflammatory processes. The aim of this study was to investigate the effect of PGI2on chemotaxis of human fetal lung fibroblasts (HFL-1). Using the blind well chamber technique, we found that the PGI2analog carbaprostacyclin (10−6M) inhibited HFL-1 chemotaxis to human plasma fibronectin (20 μg/ml) 58.0 ± 13.2% ( P < 0.05) and to platelet-derived growth factor (PDGF)-BB (10 ng/ml) 48.7 ± 4.6% ( P < 0.05). Checkerboard analysis demonstrated that carbaprostacyclin inhibits both directed and undirected migration. The inhibitory effect of the carbaprostacyclin was concentration dependent and blocked by the cAMP-dependent protein kinase (PKA) inhibitor KT-5720, suggesting that a cAMP-PKA pathway may be involved in the process. Two other PGI2analogs, ciprostene and dehydro-15-cyclohexyl carbaprostacyclin (both 10−6M), significantly inhibited fibroblast migration to fibronectin. In summary, PGI2appears to inhibit fibroblast chemotaxis to fibronectin and PDGF-BB. Such an effect may contribute to the regulation of fibroblasts in wound healing and could contribute to the pathogenesis of diseases characterized by abnormal tissue repair remodeling.

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N,N-dimethyl phytosphingosine induces caspase-8-dependent cytochrome c release and apoptosis through ROS generation in human leukemia cells

Toxicology and Applied Pharmacology ◽

10.1016/j.taap.2009.05.020 ◽

2009 ◽

Vol 239 (1) ◽

pp. 87-97 ◽

Cited By ~ 40

Author(s):

Byeong Mo Kim ◽

Yun Jung Choi ◽

Youngsoo Han ◽

Yeon-Sook Yun ◽

Sung Hee Hong

Keyword(s):

Cytochrome C ◽

Leukemia Cells ◽

Ros Generation ◽

Caspase 8 ◽

Human Leukemia ◽

Cytochrome C Release ◽

Human Leukemia Cells

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Collagen glucosyl- and galactosyltransferases of cultured human fetal lung fibroblasts

Archives of Biochemistry and Biophysics ◽

10.1016/0003-9861(81)90235-6 ◽

1981 ◽

Vol 210 (2) ◽

pp. 678-690 ◽

Cited By ~ 8

Author(s):

Hilda H. Carnicero ◽

Anthony M. Adamany ◽

Sasha Englard

Keyword(s):

Fetal Lung ◽

Lung Fibroblasts ◽

Human Fetal Lung

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Cytochrome c release from rat liver mitochondria is compromised by increased saturated cardiolipin species induced by sucrose feeding

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00617.2014 ◽

2015 ◽

Vol 309 (9) ◽

pp. E777-E786 ◽

Cited By ~ 5

Author(s):

Angélica Ruiz-Ramírez ◽

Miguel-Angel Barrios-Maya ◽

Ocarol López-Acosta ◽

Dora Molina-Ortiz ◽

Mohammed El-Hafidi

Keyword(s):

Cytochrome C ◽

Superoxide Radical ◽

Lipid Membrane ◽

Liver Mitochondria ◽

Membrane Lipid ◽

Rat Liver Mitochondria ◽

Ros Generation ◽

Cytochrome C Release ◽

Mitochondrial Inner Membrane ◽

Sucrose Feeding

Cytochrome c release from mitochondria has been described to be related to reactive oxygen species (ROS) generation. With ROS generation being increased in fatty liver from sucrose-fed (SF) rats, we hypothesized that cytochrome c release might be positively associated with H2O2 generation from SF mitochondria. Surprisingly, cytochrome c release from mitochondria of SF liver was found to be significantly lower compared with control (C) mitochondria oxidizing pyruvate/malate or succinate. Exposure of mitochondria to exogenous superoxide radical generated by the xanthine/xanthine oxidase system elicits a dose-response cytochrome c release in both control and SF mitochondria, but cytochrome c release remains lower in SF mitochondria compared with C mitochondria. Furthermore, the addition of ebselen, PEG-catalase, or catalase, a H2O2 scavenger, significantly reduces cytochrome c release from C and SF mitochondria. Our results suggest that both intra- and extramitochondrial H2O2 are involved in cytochrome c release, but the persisting difference between C and SF levels can be attributed to the differences in cardiolipin compositions. Indeed, the ratio of palmitic acid-rich cardiolipin species was found to be increased in lipid membrane from SF mitochondria compared with C mitochondria, whereas that of linoleic acid-rich cardiolipin species was found decreased. In addition, the content of tafazzin, a protein responsible for cardiolipin remodeling, was decreased in SF mitochondria. Therefore, we conclude that the changes observed in the composition of cardiolipin molecular species in SF mitochondria may be involved in cytochrome c interaction with mitochondrial inner membrane lipid and in its reduced release from SF mitochondria.

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Loss of FLCN inhibits canonical WNT signaling via TFE3

Human Molecular Genetics ◽

10.1093/hmg/ddz158 ◽

2019 ◽

Vol 28 (19) ◽

pp. 3270-3281 ◽

Cited By ~ 8

Author(s):

John C Kennedy ◽

Damir Khabibullin ◽

Thomas Hougard ◽

Julie Nijmeh ◽

Wei Shi ◽

...

Keyword(s):

Wnt Pathway ◽

Lower Lobe ◽

Fetal Lung ◽

Lung Fibroblasts ◽

Pulmonary Cysts ◽

Wnt Ligands ◽

Neonatal Lung ◽

Canonical Wnt ◽

Human Fetal Lung ◽

Bhd Syndrome

Abstract Lower lobe predominant pulmonary cysts occur in up to 90% of patients with Birt–Hogg–Dubé (BHD) syndrome, but the key pathologic cell type and signaling events driving this distinct phenotype remain elusive. Through examination of the LungMAP database, we found that folliculin (FLCN) is highly expressed in neonatal lung mesenchymal cells. Using RNA-Seq, we found that inactivation of Flcn in mouse embryonic fibroblasts leads to changes in multiple Wnt ligands, including a 2.8-fold decrease in Wnt2. This was associated with decreased TCF/LEF activity, a readout of canonical WNT activity, after treatment with a GSK3-α/β inhibitor. Similarly, FLCN deficiency in HEK293T cells decreased WNT pathway activity by 76% post-GSK3-α/β inhibition. Inactivation of FLCN in human fetal lung fibroblasts (MRC-5) led to ~ 100-fold decrease in Wnt2 expression and a 33-fold decrease in Wnt7b expression—two ligands known to be necessary for lung development. Furthermore, canonical WNT activity was decreased by 60%. Classic WNT targets such as AXIN2 and BMP4, and WNT enhanceosome members including TCF4, LEF1 and BCL9 were also decreased after GSK3-α/β inhibition. FLCN-deficient MRC-5 cells failed to upregulate LEF1 in response to GSK3-α/β inhibition. Finally, we found that a constitutively active β-catenin could only partially rescue the decreased WNT activity phenotype seen in FLCN-deficient cells, whereas silencing the transcription factor TFE3 completely reversed this phenotype. In summary, our data establish FLCN as a critical regulator of the WNT pathway via TFE3 and suggest that FLCN-dependent defects in WNT pathway developmental cues may contribute to lung cyst pathogenesis in BHD.

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