Chloroquine Enhances Death in Lung Adenocarcinoma A549 Cells Exposed to Cold Atmospheric Plasma Jet

Abstract

Cold atmospheric plasma (CAP) is an intensively-studied approach for the treatment of malignant neoplasms. Various active oxygen and nitrogen compounds are believed to be the main cytotoxic effectors on biotargets; however, the comprehensive mechanism of CAP interaction with living cells and tissues remains elusive. In this study, we experimentally determined the optimal discharge regime (or semi-selective regime) for the direct CAP jet treatment of cancer cells, under which lung adenocarcinoma A549, A427 and NCI-H23 cells demonstrated substantial suppression of viability, coupled with a weak viability decrease of healthy lung fibroblasts Wi-38 and MRC-5. The death of CAP-exposed cancer and healthy cells under semi-selective conditions was caspase-dependent. We showed that there was an accumulation of lysosomes in the treated cells. The increased activity of lysosomal protease Cathepsin D, the transcriptional upregulation of autophagy-related MAPLC3B gene in cancer cells and the changes in autophagy-related proteins may have indicated the activation of autophagy. The addition of the autophagy inhibitor chloroquine (CQ) after the CAP jet treatment increased the death of A549 cancer cells in a synergistic manner and showed a low effect on the viability of CAP-treated Wi-38 cells. Downregulation of Drp1 mitochondrial protein and upregulation of PINK1 protein in CAP + CQ treated cells indicated that CQ increased the CAP-dependent destabilization of mitochondria. We concluded that CAP weakly activated pro-survival autophagy in irradiated cells, and CQ promoted CAP-dependent cell death due to the destabilization of autophagosomes formation and mitochondria homeostasis. To summarize, the combination of CAP treatment with CQ could be useful for the development of cold plasma-based antitumor approaches for clinical application.

Personalized Plasma Medicine for Cancer: Transforming Treatment Strategies with Mathematical Modeling and Machine Learning Approaches

“…To achieve personalized treatment, CAP is emerging as a promising approach, as the parameters of the plasma device can be fine-tuned during the treatment to suit the patient's specific dosage. CAP has been shown to reprogram the tumor microenvironment [143][144][145], inhibit tumor growth with high selectivity, and even enhance the effectiveness of other treatment modalities like chemotherapy [146][147][148][149], immunotherapy [150,151], radiotherapy [152][153][154][155], and nanomedicine [156][157][158][159][160] with improved treatment outcomes and minimized sideeffects [161]. The self-adaptive nature of the adaptive plasma system complements the power of ML.…”

Pulsed voltage cold atmospheric plasma jet and gold nanoparticles enhance cytotoxic anticancer effect

“…In order to find out a time range of cell membrane permeability for NP uptake, we added NP-PEG-FAM to the cells 0-4 h after CAPJ treatment. Next, we focused on lung tumor cells because we have already shown molecular features of cell death under the action of CAPJ on these cell lines [41,44], and this study is a logical progression of the previous study. The use of A549 and H23 cell lines will allow us to further compare whether the molecular mechanism of cell death is altered by the addition of NPs.…”

“…NCI-H23 epithelial-like lung adenocarcinoma cells (ATCC # CRL-5800) were grown in RPMI 1640 (Gibco, USA) supplemented with 10% fetal bovine serum (Sigma-Aldrich), 2 mM L-glutamine, 250 mg ml −1 amphotericin B and 100 U ml −1 penicillin/streptomycin in a humidified atmosphere containing 5% CO 2 at 37 • C. uMel1 cells were grown in α-MEM medium (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 20% fetal bovine serum (GIBCO, Thermo Fisher, Waltham, MA, USA), 1x GlutaMAX TM , 250 mg ml −1 amphotericin B and 100 U ml −1 penicillin/streptomycinc, 5 µg ml −1 insulin (SCI-store, Moscow, Russia), 20 ng ml −1 EGF (SCI-store, Moscow, Russia), 1x MITO (BD Biosciences-Discovery Labware, San Jose, CA, USA). A549 and H23 cells were maintained as previously described [41]. BrCCh4e-134 cells were cultivated as previously described [40].…”

“…The cells that had reached 30% confluence in a 96-well plate (TPP, Trasadingen, Switzerland) were treated with CAPJ or NPs and their combination. Cell viability was detected 24 h after CAPJ irradiation using a MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) test as was described previously [41,42].…”

EGFR Suppression Inhibits the Sphere Formation of MCF7 Cells Overexpressing EGFR

“…Earlier, we obtained an MCF7 human breast adenocarcinoma cell line that manifested an increased expression of EGFR. The study revealed that excessive EGFR in MCF7 cells led to the spontaneous formation of spheres under standard culture conditions [ 10 , 11 ]. MCF7-EGFR spheroids have a round shape with a well-defined outer boundary and a median diameter of 100 μm, with the size of large spheroids likely to exceed 400 μm.…”