Mono-2-ethylhexyl phthalate induced loss of mitochondrial membrane potential and activation of Caspase3 in HepG2 cells

2012 ◽  
Vol 33 (3) ◽  
pp. 421-430 ◽  
Author(s):  
Xi Chen ◽  
Jianshu Wang ◽  
Qizhi Qin ◽  
Ying Jiang ◽  
Guangtao Yang ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Ethylhexyl Phthalate

Encircling granulosa cells protects against di-(2-ethylhexyl)phthalate-induced apoptosis in rat oocytes cultured in vitro

Zygote ◽  
2019 ◽  
Vol 27 (4) ◽  
pp. 203-213 ◽  
Author(s):  
Anima Tripathi ◽  
Vivek Pandey ◽  
A.N. Sahu ◽  
Alok K. Singh ◽  
Pawan K. Dubey
Keyword(s):  
Oxidative Stress ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Granulosa Cells ◽  
Mitochondrial Membrane ◽  
Dependent Manner ◽  
Apoptotic Markers ◽  
Induced Apoptosis ◽  
Ethylhexyl Phthalate

SummaryThe present study investigated if the presence of encircling granulosa cells protected against di(2-ethylhexyl)phthalate (DEHP)-induced oxidative stress in rat oocytes cultured in vitro. Denuded oocytes and cumulus–oocyte complexes (COCs) were treated with or without various doses of DEHP (0.0, 25.0, 50.0, 100, 200, 400 and 800 μM) in vitro. Morphological apoptotic changes, levels of oxidative stress and reactive oxygen species (ROS), mitochondrial membrane potential, and expression levels of apoptotic markers (Bcl2, Bax, cytochrome c) were analyzed. Our results showed that DEHP induced morphological apoptotic changes in a dose-dependent manner in denuded oocytes cultured in vitro. The effective dose of DEHP (400 µg) significantly (P>0.05) increased oxidative stress by elevating ROS levels and the mitochondrial membrane potential with higher mRNA expression and protein levels of apoptotic markers (Bax, cytochrome c). Encircling granulosa cells protected oocytes from DEHP-induced morphological changes, increased oxidative stress and ROS levels, as well as increased expression of apoptotic markers. Taken together our data suggested that encircling granulosa cells protected oocytes against DEHP-induced apoptosis and that the presence of granulosa cells could act positively towards the survival of oocytes under in vitro culture conditions and may be helpful during assisted reproductive technique programmes.

Lysosome Inhibitors Enhance the Chemotherapeutic Activity of Doxorubicin in HepG2 Cells

Chemotherapy ◽  
2016 ◽  
Vol 62 (2) ◽  
pp. 85-93 ◽  
Author(s):  
Yuyin Li ◽  
Yuejun Sun ◽  
Lifang Jing ◽  
Jianjun Wang ◽  
Yali Yan ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Inhibitory Effect ◽  
Membrane Permeabilization ◽  
Lysosomal Membrane ◽  
Bafilomycin A1 ◽  
Lysosomal Membrane Permeabilization ◽  
Proliferative Ability

The lysosome inhibitors bafilomycin A1 and chloroquine have both lysosomotropic properties and autophagy inhibition ability, and are promising clinical agents to be used in combination with anticancer drugs. In order to investigate this combination effect, HepG2 cells were treated with bafilomycin A1, chloroquine, or/and doxorubicin, and their proliferative ability, induction of apoptosis, and the changes of lysosomal membrane permeabilization and mitochondrial membrane potential were studied. The results demonstrate that treatment with bafilomycin A1 or chloroquine alone at a relatively low concentration promotes the inhibitory effect of doxorubicin on cell growth and apoptosis. Further studies reveal that bafilomycin A1 and chloroquine promote lysosomal membrane permeabilization and the reduction of mitochondrial membrane potential induced by doxorubicin. Our findings suggest that bafilomycin A1 and chloroquine potentiate the anticancer effect of doxorubicin in hepatic cancer cells and that supplementation of conventional chemotherapy with lysosome inhibitors may provide a more efficient anticancer therapy.

Study on Capparis spionosa L. Polysaccharide (CSPS) Induced HepG2 Apoptosis by Controlling Ca2+ Path

2011 ◽  
Vol 282-283 ◽  
pp. 203-208 ◽  
Author(s):  
Yu Bin Ji ◽  
Fang Dong ◽  
Shi Yong Gao ◽  
Miao Yu
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Hepatoma Cell ◽  
Scanning Microscope ◽  
Human Hepatoma Cell ◽  
Bax Protein ◽  
Confocal Scanning ◽  
Labeling Method

To investigate the mechanism on Capparis spionosa L polysaccharide(CSPS) inducing apoptosis in HepG2 human hepatoma cell. MTT was ddopted to determine if CSPS had cytotoxic effect on HepG2. Morphology of HepG2 changed with dosages of CSPS was detected by laser confocal scanning microscope. Flow cytometry(FCM) was used to detect the apoptosis by PI labeling method. Calcium, mitochondrial membrane potential, Bcl-2/Bax of HepG2 cells were detected by laser confocal scanning microscope. The result of MTT showed that CSPS could inhibit the growth of HepG2 significantly. HepG2 cells were shrinkage, fragmentation, appearance of apoptotic bodies by laser confocal scanning microscope. HepG2 cell apoptosis rate was increased gradually with dosage by FCM. Calcium concentration, mitochondrial membrane potential, Bcl-2 protein of HepG2 were decreased, Bax protein content of HepG2 was increased by laser confocal scanning microscope. CSPS induced HepG2 apoptosis by controlling Bax/Bcl-2 in Ca2+ path.

scholarly journals Mitochondrial membrane potential played crucial roles in the accumulation of berberine in HepG2 cells

2019 ◽  
Vol 39 (4) ◽  
Author(s):  
Qiao Li ◽  
Ting Zhou ◽  
Chang Liu ◽  
Xiao-Yu Wang ◽  
Ji-Quan Zhang ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Cell Membrane ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Laser Scanning ◽  
Laser Scanning Confocal Microscopy ◽  
Cell Membrane Potential ◽  
Cell Organelles ◽  
Chromatography Method

Abstract Berberine is a natural alkaloid that has antineoplastic effects. However, in hepatoma cells like HepG2, the expressions of uptake transporters are minimal but efflux transporters are relatively high. Hence, how berberine enters and reaches a cytocidal concentration remains to be elucidated. In the present study, we revealed the accumulation mechanism of berberine in HepG2 cells. Cell organelles were isolated based on differential centrifugation; berberine concentration was measured using a liquid chromatography-tandem mass chromatography method or flow cytometry. Subcellular distribution of berberine was observed using a laser scanning confocal microscopy. The results showed that berberine was concentration-, temperature-, and time-dependently taken up and accumulated in HepG2 cells. Membrane drug transporters and cell membrane potential had limited effects in berberine uptake. However, qualitative and quantitative studies showed that berberine was enriched in the mitochondria; inhibition of mitochondrial membrane potential (MMP) by carbonyl cyanide 3-chlorophenylhydrazone (CCCP) significantly decreased the intracellular berberine by up to 70%. More importantly, MMP not only significantly enhanced berberine uptake driven by cell membrane potential (P<0.01) but also inhibited p-glycoprotein (P-gp)-mediated berberine efflux (P<0.01). In brief, our results for the first time showed that MMP played crucial roles in berberine accumulation in HepG2 cells.

scholarly journals Antiproliferative activity of Syzygium coriaceum, an endemic plant of Mauritius, with its UPLC-MS metabolite fingerprint: A mechanistic study

PLoS ONE ◽  
2021 ◽  
Vol 16 (6) ◽  
pp. e0252276
Author(s):  
Nawraj Rummun ◽  
Ahmed Serag ◽  
Philippe Rondeau ◽  
Srishti Ramsaha ◽  
Emmanuel Bourdon ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Untreated Control ◽  
Metabolite Profiling ◽  
Mitochondrial Membrane ◽  
Leaf Extract ◽  
Biological Activities ◽  
Mechanistic Study ◽  
Endemic Plant

Flowering plants from the Syzygium genus have long been used in different ethnomedicinal systems worldwide and have been under scrutiny for their biological activities. Syzygium coriaceum, an endemic plant of Mauritius has been poorly studied for its potential application against cancer. Herein, Syzygium coriaceum leaf extract has been investigated for its anticancer effect against hepatocellular carcinoma (HepG2) cells. The anticancer activity was assessed using cell proliferation assays, flow cytometry, JC-1 mitochondrial membrane potential assay, and the COMET assay. Un-targeted metabolite profiling via ultra-performance liquid chromatography coupled to high-resolution qTOF-MS (UPLC-MS) and aided by molecular networking was employed to identify the crude extract metabolites. S. coriaceum treatment induced a dose-dependent increase in lactate dehydrogenase leakage into the culture media, peaking up to 47% (p ≤ 0.0001), compared to untreated control. Moreover, at 40 μg/mL, S. coriaceum led to 88.1% (p ≤ 0.0001) drop in mitochondrial membrane potential and 5.7% (p ≤ 0.001) increased in the number of the cell population in G0/G1 phase as well as increased (p < 0.05) the proportion of cells undergoing apoptotic/necrotic cell death. More so, at 10 μg/mL, S. coriaceum induced DNA damage which was 19 folds (p < 0.001) higher than that of untreated control cells. Metabolite profiling indicated the presence of 65 metabolites, out of which 59 were identified. Tannins, flavonoids, nitrogenous compounds, and organic acids were the most predominant classes of compounds detected. Our findings showed that the presence of tannins and flavonoids in S. coriaceum leaf extract could account for the multiple mechanisms of actions underlying the antiproliferative effect against HepG2 cells.

scholarly journals Sub-Lethal Hyperthermia Enhances Anticancer Activity of Doxorubicin in Chronically Hypoxic HepG2 Cells Through ROS-Dependent Mechanism

2020 ◽  
Author(s):  
Qi Wang ◽  
Hui Zhang ◽  
Qianqian Ren ◽  
Tianhe Ye ◽  
Yiming Liu ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Cell Viability ◽  
Anticancer Activity ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Thermal Ablation ◽  
Mitochondrial Membrane ◽  
Intracellular Uptake ◽  
Underlying Mechanisms ◽  
Dependent Mechanism

Abstract Background and aims: Thermal ablation in combination with transarterial chemoembolization (TACE) has been reported to exert a more powerful anti-tumor effect than thermal ablation alone in hepatocellular carcinoma (HCC) patients. However, the underlying mechanisms remain unclear. The purpose of this study was to evaluate whether sub-lethal hyperthermia encountered in the peri-ablation zone during thermal ablation enhances the anticancer activity of doxorubicin in chronically hypoxic (encountered in the tumor area after TACE) liver cancer cells and to explore the underlying mechanisms. Methods HepG2 cells pre-cultured under chronic hypoxic conditions (1% oxygen) were treated in a 42 °C water bath for 15 min or 30 min, followed by incubation with doxorubicin. Assays were then performed to determine intracellular uptake of doxorubicin, cell viability, apoptosis, cell cycle, mitochondrial membrane potential (MMP), reactive oxygen species (ROS), and total antioxidant capacity. Results The results confirmed that sub-lethal hyperthermia enhanced intracellular uptake of doxorubicin into hypoxic HepG2 cells. Hyperthermia combined with doxorubicin led to a greater inhibition of cell viability and increased apoptosis in hypoxic HepG2 cells compared to hyperthermia or doxorubicin alone. In addition, the combination induced apoptosis by increasing ROS and causing disruption of mitochondrial membrane potential. Pretreatment with the ROS scavenger N-acetyl cysteine (NAC) significantly inhibited the apoptotic response suggesting that cell death is ROS-dependent. Conclusions These findings suggest that sub-lethal hyperthermia enhanced the anti-cancer activity of doxorubicin in hypoxic HepG2 cells through ROS-dependent mechanism.

scholarly journals The effects and mechanism of peiminine-induced apoptosis in human hepatocellular carcinoma HepG2 cells

2018 ◽  
Author(s):  
Xu Chao ◽  
Guoquan Wang ◽  
Yuping Tang ◽  
Changhu Dong ◽  
Hong Li ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Flow Cytometry ◽  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Hepg2 Cells ◽  
Mitochondrial Membrane ◽  
Molecular Mechanisms ◽  
Human Hepatocellular Carcinoma ◽  
Nuclear Staining ◽  
Induced Apoptosis

AbstractPeiminine is a compound that is isolated fromBolbostemma paniculatum(Maxim) Franquet (Cucurbitaceae family), which has demonstrated antitumor activities. Its precise molecular mechanisms underlying antitumor activity remain elusive. In this study, peiminine-induced apoptosis towards human hepatocellular carcinoma and its molecular mechanisms were investigated. MTT assay was employed to assess anticancer effects of peiminine at concentrations of 2, 4, 6, 8, 10, 12, and 14 μg/ml after 24, 48, or 72 h. Nuclear staining and flow cytometry were carried out to further assess apoptosis. Mitochondrial membrane potential evaluation and Western blot analysis were performed to investigate the mechanism of peiminine-induced apoptosis. Peiminine reduced the viability of HepG2 cells in a time- and dose-dependent manner and had an IC50of 4.58 μg/mL at 24h. Flow cytometry assessment indicated that peiminine markedly increased the cell number of apoptotic cells and the mitochondrial membrane potential dose-dependently in HepG2 cells. The results of Western blotting showed the expression of Bcl-2, procaspase-3, procaspase-8, procaspase-9, and PARP1decreased in HepG2 cells treated with peiminine, while the expression of Bax, caspase-3, caspase-8, caspase-9, and cleaved PARP1increased. The result suggest taht peiminine can induce apoptosis in human hepatocellular carcinoma HepG2 cells through both extrinsic and intrinsic apoptotic pathways.

Sign in / Sign up

Export Citation Format

Share Document