Autophagy and Apoptosis Induced in U87 MG Glioblastoma Cells by Hypericin-Mediated Photodynamic Therapy Can Be Photobiomodulated with 808 nm Light

Pevná, Viktória; Wagnières, Georges; Huntošová, Veronika

doi:10.3390/biomedicines9111703

Cited by 19 publications

(24 citation statements)

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“…CRT exposure on HT-29 cell surfaces was therefore evaluated 6 h after Hyp-SDT treatment, and a significant increase in the CRT fluorescent signal was observed in the SDT condition compared with untreated cells (Ctrl), which are indicated by a dashed line ( Figure 7 , p ≤ 0.05). Another important ICD hallmark is the release of HMGB1 from the nucleus of dying cells, and this is considered a late signal in the ICD pathway [ 40 ]. Therefore, HMGB1 release was evaluated in HT-29 cells 24 h after Hyp-SDT treatment.…”

Section: Resultsmentioning

confidence: 99%

“…At the end of each incubation time, cells were detached using trypsin, centrifuged, and resuspended in 100 µL of PBS. Cells were then analyzed using flow cytometry (Accuri C6, Milano, Italy), with excitation at 488 nm, in the spectral range >590 nm [ 40 ], considering 1 × 10 4 events. Cells debris with low forward scatter (FSC) and side scatter (SSC) was excluded from the analysis.…”

Section: Methodsmentioning

confidence: 99%

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Ultrasound Triggers Hypericin Activation Leading to Multifaceted Anticancer Activity

et al. 2022

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The use of ultrasound (US) in combination with a responsive chemical agent (sonosensitizer) can selectively trigger the agent’s anticancer activity in a process called sonodynamic therapy (SDT). SDT shares some properties with photodynamic therapy (PDT), which has been clinically approved, but sets itself apart because of its use of US rather than light to achieve better tissue penetration. SDT provides anticancer effects mainly via the sonosensitizer-mediated generation of reactive oxygen species (ROS), although the precise nature of the underpinning mechanism is still under debate. This work investigates the SDT anticancer activity of hypericin (Hyp) in vitro in two- (2D) and three-dimensional (3D) HT-29 colon cancer models, and uses PDT as a yardstick due to its well-known Hyp phototoxicity. The cancer cell uptake and cellular localization of Hyp were investigated first to determine the proper noncytotoxic concentration and incubation time of Hyp for SDT. Furthermore, ROS production, cell proliferation, and cell death were evaluated after Hyp was exposed to US. Since cancer relapse and transporter-mediated multidrug resistance (MDR) are important causes of cancer treatment failure, the US-mediated ability of Hyp to elicit immunogenic cell death (ICD) and overcome MDR was also investigated. SDT showed strong ROS-mediated anticancer activity 48 h after treatment in both the HT-29 models. Specific damage-associated molecular patterns that are consistent with ICD, such as calreticulin (CRT) exposure and high-mobility group box 1 protein (HMGB1) release, were observed after SDT with Hyp. Moreover, the expression of the ABC transporter, P-glycoprotein (P-gp), in HT-29/MDR cells was not able to hinder cancer cell responsiveness to SDT with Hyp. This work reveals, for the first time, the US responsiveness of Hyp with significant anticancer activity being displayed, making it a full-fledged sonosensitizer for the SDT of cancer.

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Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Ultrasound Triggers Hypericin Activation Leading to Multifaceted Anticancer Activity

et al. 2022

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“…The response of the cells to PDT was determined 24 h after PDT. The optimization of the therapeutic protocol was based on our previous publication, in which the control measurements were performed without hypericin and light [ 12 , 24 ]. No significant effects of light application were observed in the studied cells.…”

Section: Methodsmentioning

confidence: 99%

“…Hypericin, a natural compound, has been presented as a very potent photosensitizer that induces PDT via singlet oxygen generation [ 9 ]. It has been shown to induce apoptosis in several cell lines, with p38, Jun N terminal kinase, caspase-9, and caspase-3 playing important roles [ 10 , 11 , 12 , 13 ].…”

Section: Introductionmentioning

confidence: 99%

Spheroidal Model of SKBR3 and U87MG Cancer Cells for Live Imaging of Caspase-3 during Apoptosis Induced by Singlet Oxygen in Photodynamic Therapy

et al. 2022

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Aspects related to the response of cells to photodynamic therapy (PDT) have been well studied in cell cultures, which often grow in monolayers. In this work, we propose a spheroidal model of U87MG and SKBR3 cells designed to mimic superficial tumor tissue, small spheroids (<500 µm) suitable for confocal fluorescence microscopy, and larger spheroids (>500 µm) that can be xenografted onto quail chorioallantoic membrane (CAM) to study the effects of PDT in real time. Hypericin was used as a model molecule for a hydrophobic photosensitizer that can produce singlet oxygen (1O2). 1O2 production by hypericin was detected in SKBR3 and U87MG spheroid models using a label-free technique. Vital fluorescence microscopy and flow cytometry revealed the heterogeneity of caspase-3 distribution in the cells of the spheroids. The levels of caspase-3 and apoptosis increased in the cells of spheroids 24 h after PDT. Lactate dehydrogenase activity was evaluated in the spheroids as the most reliable assay to detect differences in phototoxicity. Finally, we demonstrated the applicability of U87MG spheroids on CAM in photodiagnostics. Overall, the variability and applicability of the prepared spheroid models were demonstrated in the PDT study.

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“…During the early phase of cerebral ischemia, injury stimulates the differentiation of astrocytes, which protect the survival of neurons and maintain the normal function of the central nervous system. Astrocytes play an important role in neuroregulation and synaptic transmission [ 13 ], and human U87 astrocytes were employed as a cellular model in this study [ 14 ]. Therefore, astrocytes involved in autophagy through the PI3K/Akt/mTOR signaling pathway may be a potential treatment for ischemic stroke.…”

Section: Introductionmentioning

confidence: 99%

Cornin protects against cerebral ischemia/reperfusion injury by preventing autophagy via the PI3K/Akt/mTOR pathway

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et al. 2022

BMC Pharmacol Toxicol

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Background Ischemia stroke is the leading cause of disability, which is a consequence of vascular occlusion. The purpose of this study is to investigate the effect of cornin which is isolated from the fruit of Verbena officinalis L, against astrocytes autophagy induced by cerebral ischemia/reperfusion (CI/R) injury in vitro and in vivo and its potential mechanism. Methods Cornin at dose of 2.5, 5 and 10 mg/kg were intravenously injected to MCAO rats at 15 min after reperfusion. The infarction volume, blood–brain barrier (BBB), neurological severity score (mNSS), and autophagy related protein were used to evaluated the protective effects and potential mechanism of cornin in autophagy with or without phosphoinositide-3 kinase (PI3K)inhibitor LY294002 and mammalian target of rapamycin (mTOR) small interfering RNA (siRNA) at 24 h after CI/R injury. The potential protective effects and mechanism of cornin at concention of 10 ~ 1000 nM were also evaluated in oxygen glucose deprivation/reperfusion (OGD/R) in U87 cells. Results The results suggest that cornin at dose of 5 or 10 mg/kg significantly reduce the cerebral infarction volume and blood–brain barrier (BBB) leakage, and improve neurological recovery in MCAO rats. Cleaved caspase-3 and Bax levels were significantly decreased, while B-cell lymphoma-2 (Bcl-2) and the apoptosis regulator ratio (Bcl-2/Bax) were markedly increased when treated with 2.5–10 mg/kg cornin. The obvious decreased expressions of glial fibrillary acidic protein (GFAP), myosin-like BCL2 interacting protein (Beclin-1) and microtubule-associated protein light chain 3 II (LC3-II) and increased of neuronal nuclei (NeuN), sequestosome-1 (p62), phosphorylated mTOR (p-mTOR), and phosphorylated Akt (p-Akt) were observed in MCAO rats treated with 10 mg/kg cornin, which was counteracted by LY294002. The expression of autophagy-related proteins with or without LY294002 and mTOR siRNA presented the similar results as in vitro in OGD/R in U87 cells. Conclusions These results indicate that cornin improved neurological recovery after cerebral ischemia injury by preventing astrocytes autophagy induced by CI/R via the PI3K/Akt/mTOR signaling pathway.

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Autophagy and Apoptosis Induced in U87 MG Glioblastoma Cells by Hypericin-Mediated Photodynamic Therapy Can Be Photobiomodulated with 808 nm Light

Cited by 19 publications

References 50 publications

Ultrasound Triggers Hypericin Activation Leading to Multifaceted Anticancer Activity

Ultrasound Triggers Hypericin Activation Leading to Multifaceted Anticancer Activity

Spheroidal Model of SKBR3 and U87MG Cancer Cells for Live Imaging of Caspase-3 during Apoptosis Induced by Singlet Oxygen in Photodynamic Therapy

Cornin protects against cerebral ischemia/reperfusion injury by preventing autophagy via the PI3K/Akt/mTOR pathway

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