TY - JOUR

T1 - Chloroquine Enhances Death in Lung Adenocarcinoma A549 Cells Exposed to Cold Atmospheric Plasma Jet

AU - Patrakova, Ekaterina

AU - Biryukov, Mikhail

AU - Troitskaya, Olga

AU - Gugin, Pavel

AU - Milakhina, Elena

AU - Semenov, Dmitriy

AU - Poletaeva, Julia

AU - Ryabchikova, Elena

AU - Novak, Diana

AU - Kryachkova, Nadezhda

AU - Polyakova, Alina

AU - Zhilnikova, Maria

AU - Zakrevsky, Dmitriy

AU - Schweigert, Irina

AU - Koval, Olga

N1 - Funding: This research was funded by Russian Science Foundation, grant number 19-19-00255П.

PY - 2023/1/12

Y1 - 2023/1/12

N2 - 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.

AB - 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.

KW - Humans

KW - Chloroquine/pharmacology

KW - A549 Cells

KW - Plasma Gases/pharmacology

KW - Apoptosis

KW - Adenocarcinoma of Lung/drug therapy

KW - Lung Neoplasms/drug therapy

UR - https://www.scopus.com/record/display.uri?eid=2-s2.0-85146824903&origin=inward&txGid=6ebc7ded923600c28c6f69babf8dc1c1

UR - https://www.mendeley.com/catalogue/5936394e-c069-3c98-a956-4274eac99ab5/

U2 - 10.3390/cells12020290

DO - 10.3390/cells12020290

M3 - Article

C2 - 36672225

VL - 12

JO - Cells

JF - Cells

SN - 2073-4409

IS - 2

M1 - 290

ER -