The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment Strategy

<b>Background/Objectives</b>: Cold atmospheric plasma (CAP) has been demonstrated as an adjustable device to generate various combinations of short-lived reactive oxygen and nitrogen species (RONS) and as a promising appliance for cancer therapy. This study investigated the effects of di...

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Main Authors: Chao-Yu Chen, Chung-Hsien Chou, Yun-Chien Cheng
Format: Article
Language:English
Published: MDPI AG 2025-01-01
Series:Biomedicines
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Online Access:https://www.mdpi.com/2227-9059/13/1/184
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author Chao-Yu Chen
Chung-Hsien Chou
Yun-Chien Cheng
author_facet Chao-Yu Chen
Chung-Hsien Chou
Yun-Chien Cheng
author_sort Chao-Yu Chen
collection DOAJ
description <b>Background/Objectives</b>: Cold atmospheric plasma (CAP) has been demonstrated as an adjustable device to generate various combinations of short-lived reactive oxygen and nitrogen species (RONS) and as a promising appliance for cancer therapy. This study investigated the effects of direct and indirect treatments of Argon-based CAP to cancer cells (A2058, A549, U2OS and BCC) and fibroblasts (NIH3T3 and L929) on cell viability. We also aimed to understand whether plasma-generated RONS were involved in this process using genetic evidence. <b>Methods</b>: The intensity of reactive species in the plasma gas and the concentrations of RONS in phosphate-buffered saline (PBS) and cell culture medium were measured. A viability assay was performed after the cells were treated by plasma in PBS and medium with various volumes to realize the lethal effects of plasma under different conditions. Diverse cells were treated in the same solution to compare the sensitivities of different cells to plasma treatments. The gene expression profiles of A2058 cells after the direct and indirect treatments were analyzed by next generation gene sequencing. Accordingly, we discovered the advantages of sequential treatments on cancer therapy. <b>Results</b>: The cumulative concentration of hydroxyterephthalic acid (HTA) revealed that the pre-existing OH radical (•OH) in PBS increased with the treatment durations. However, there was no significant increase in the concentration of HTA in culture medium. HTA was detected in the treatment interface of PBS but not medium, showing the penetration of •OH through PBS. The concentrations of H<sub>2</sub>O<sub>2</sub> and NO<sub>2</sub><sup>−</sup> increased with the treatment durations, but that of NO<sub>3</sub><sup>−</sup> was low. The direct treatments caused stronger lethal effects on cancer cells under certain conditions. The fibroblasts showed higher tolerance to plasma treatments. From gene expression analysis, the initial observations showed that both treatments influenced transcription-related pathways and exhibited shared or unique cellular stress responses. The pre-treatments, especially of direct exposure, revealed better cancer inhibition. <b>Conclusions</b>: The anti-cancer efficiency of plasma could be enhanced by pre-treatments and by adjusting the liquid interfaces to avoid the rapid consumption of short-lived RONS in the medium. To achieve better therapeutic effects and selectivity, more evidence is necessary to find optional plasma treatments.
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spelling doaj-art-bdb01ef7203e430a9b2ce6ef82945ec62025-01-24T13:24:18ZengMDPI AGBiomedicines2227-90592025-01-0113118410.3390/biomedicines13010184The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment StrategyChao-Yu Chen0Chung-Hsien Chou1Yun-Chien Cheng2Department of Mechanical Engineering, National Yang Ming Chiao Tung University, Hsinchu 300, TaiwanInstitute of Oral Biology, National Yang Ming Chiao Tung University, Taipei 112, TaiwanDepartment of Mechanical Engineering, National Yang Ming Chiao Tung University, Hsinchu 300, Taiwan<b>Background/Objectives</b>: Cold atmospheric plasma (CAP) has been demonstrated as an adjustable device to generate various combinations of short-lived reactive oxygen and nitrogen species (RONS) and as a promising appliance for cancer therapy. This study investigated the effects of direct and indirect treatments of Argon-based CAP to cancer cells (A2058, A549, U2OS and BCC) and fibroblasts (NIH3T3 and L929) on cell viability. We also aimed to understand whether plasma-generated RONS were involved in this process using genetic evidence. <b>Methods</b>: The intensity of reactive species in the plasma gas and the concentrations of RONS in phosphate-buffered saline (PBS) and cell culture medium were measured. A viability assay was performed after the cells were treated by plasma in PBS and medium with various volumes to realize the lethal effects of plasma under different conditions. Diverse cells were treated in the same solution to compare the sensitivities of different cells to plasma treatments. The gene expression profiles of A2058 cells after the direct and indirect treatments were analyzed by next generation gene sequencing. Accordingly, we discovered the advantages of sequential treatments on cancer therapy. <b>Results</b>: The cumulative concentration of hydroxyterephthalic acid (HTA) revealed that the pre-existing OH radical (•OH) in PBS increased with the treatment durations. However, there was no significant increase in the concentration of HTA in culture medium. HTA was detected in the treatment interface of PBS but not medium, showing the penetration of •OH through PBS. The concentrations of H<sub>2</sub>O<sub>2</sub> and NO<sub>2</sub><sup>−</sup> increased with the treatment durations, but that of NO<sub>3</sub><sup>−</sup> was low. The direct treatments caused stronger lethal effects on cancer cells under certain conditions. The fibroblasts showed higher tolerance to plasma treatments. From gene expression analysis, the initial observations showed that both treatments influenced transcription-related pathways and exhibited shared or unique cellular stress responses. The pre-treatments, especially of direct exposure, revealed better cancer inhibition. <b>Conclusions</b>: The anti-cancer efficiency of plasma could be enhanced by pre-treatments and by adjusting the liquid interfaces to avoid the rapid consumption of short-lived RONS in the medium. To achieve better therapeutic effects and selectivity, more evidence is necessary to find optional plasma treatments.https://www.mdpi.com/2227-9059/13/1/184cold atmospheric plasmareactive oxygen nitrogen speciescell viabilitygene expressioncancer therapy
spellingShingle Chao-Yu Chen
Chung-Hsien Chou
Yun-Chien Cheng
The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment Strategy
Biomedicines
cold atmospheric plasma
reactive oxygen nitrogen species
cell viability
gene expression
cancer therapy
title The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment Strategy
title_full The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment Strategy
title_fullStr The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment Strategy
title_full_unstemmed The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment Strategy
title_short The Genetic Expression Difference of A2058 Cells Treated by Plasma Direct Exposure and Plasma-Treated Medium and the Appropriate Treatment Strategy
title_sort genetic expression difference of a2058 cells treated by plasma direct exposure and plasma treated medium and the appropriate treatment strategy
topic cold atmospheric plasma
reactive oxygen nitrogen species
cell viability
gene expression
cancer therapy
url https://www.mdpi.com/2227-9059/13/1/184
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