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.

A facile composite nanoparticle promoted by photoelectron transfer and consumption for tumor combination therapy

2020 ◽  
Vol 4 (10) ◽  
pp. 3047-3056
Author(s):  
Yahui Zhang ◽  
Weizhou Sha ◽  
Yang Liu ◽  
Wei Wang ◽  
Zhi Yuan
Keyword(s):  
Membrane Potential ◽  
Combination Therapy ◽  
Cancer Cells ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Membrane Permeabilization ◽  
Lysosomal Membrane ◽  
Lysosomal Membrane Permeabilization ◽  
Composite Nanoparticle ◽  
Mitochondrial Death Pathway

BTCu NPs can cause significant lysosomal membrane permeabilization (LMP) and mitochondrial membrane potential depolarization, thus indicating a lysosomal–mitochondrial death pathway in cancer cells.

scholarly journals Contribution of Particle-Induced Lysosomal Membrane Hyperpolarization to Lysosomal Membrane Permeabilization

2021 ◽  
Vol 22 (5) ◽  
pp. 2277
Author(s):  
Tahereh Ziglari ◽  
Zifan Wang ◽  
Andrij Holian
Keyword(s):  
Membrane Potential ◽  
Time Course ◽  
Causal Relation ◽  
Underlying Mechanism ◽  
Membrane Permeabilization ◽  
Lysosomal Membrane ◽  
Early Event ◽  
Inflammasome Activation ◽  
Membrane Hyperpolarization ◽  
Lysosomal Membrane Permeabilization

Lysosomal membrane permeabilization (LMP) has been proposed to precede nanoparticle-induced macrophage injury and NLRP3 inflammasome activation; however, the underlying mechanism(s) of LMP is unknown. We propose that nanoparticle-induced lysosomal hyperpolarization triggers LMP. In this study, a rapid non-invasive method was used to measure changes in lysosomal membrane potential of murine alveolar macrophages (AM) in response to a series of nanoparticles (ZnO, TiO2, and CeO2). Crystalline SiO2 (micron-sized) was used as a positive control. Changes in cytosolic potassium were measured using Asante potassium green 2. The results demonstrated that ZnO or SiO2 hyperpolarized the lysosomal membrane and decreased cytosolic potassium, suggesting increased lysosome permeability to potassium. Time-course experiments revealed that lysosomal hyperpolarization was an early event leading to LMP, NLRP3 activation, and cell death. In contrast, TiO2- or valinomycin-treated AM did not cause LMP unless high doses led to lysosomal hyperpolarization. Neither lysosomal hyperpolarization nor LMP was observed in CeO2-treated AM. These results suggested that a threshold of lysosomal membrane potential must be exceeded to cause LMP. Furthermore, inhibition of lysosomal hyperpolarization with Bafilomycin A1 blocked LMP and NLRP3 activation, suggesting a causal relation between lysosomal hyperpolarization and LMP.

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.

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