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Preferential killing of human lung cancer cell lines with mitochondrial dysfunction by nonthermal dielectric barrier discharge plasma.

Panngom K, Baik KY, Nam MK, Han JH, Rhim H, Choi EH - Cell Death Dis (2013)

Bottom Line: It is found that the cell number of lung cancer cells has been reduced more than that of the lung normal cells.The mitochondrial vulnerability to reactive species in H460 may induce distinctively selective responses.Differential mitochondrial membrane potential decrease, mitochondrial enzymatic dysfunction, and mitochondrial morphological alteration are exhibited in two cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Plasma Bioscience and Display, Kwangwoon University, Seoul, Republic of Korea.

ABSTRACT
The distinctive cellular and mitochondrial dysfunctions of two human lung cancer cell lines (H460 and HCC1588) from two human lung normal cell lines (MRC5 and L132) have been studied by dielectric barrier discharge (DBD) plasma treatment. This cytotoxicity is exposure time-dependent, which is strongly mediated by the large amount of H2O2 and NOx in culture media generated by DBD nonthermal plasma. It is found that the cell number of lung cancer cells has been reduced more than that of the lung normal cells. The mitochondrial vulnerability to reactive species in H460 may induce distinctively selective responses. Differential mitochondrial membrane potential decrease, mitochondrial enzymatic dysfunction, and mitochondrial morphological alteration are exhibited in two cell lines. These results suggest the nonthermal plasma treatment as an efficacious modality in lung cancer therapy.

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Related in: MedlinePlus

The flow cytometry data of Annexin V and 7AAD staining of lung cells after plasma treatment. (a) The normalized cell number distribution to Annexin V intensity of H460 and MRC5 was plotted by plasma exposure time. H2O2 (1 mM) was used to make positive control. (b) Apoptosis of H460 and L132 cells was evaluated after 12 and 24 h incubation from nonthermal plasma treatment for 5 min. The number represents the percentage of cells in each quadruple
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fig4: The flow cytometry data of Annexin V and 7AAD staining of lung cells after plasma treatment. (a) The normalized cell number distribution to Annexin V intensity of H460 and MRC5 was plotted by plasma exposure time. H2O2 (1 mM) was used to make positive control. (b) Apoptosis of H460 and L132 cells was evaluated after 12 and 24 h incubation from nonthermal plasma treatment for 5 min. The number represents the percentage of cells in each quadruple

Mentions: To check whether cell death induced by plasma is apoptosis, we stained cells with Annexin V, an early apoptotic indicator. In Figure 4a, enhanced fluorescent intensity of Annexin V shows the increase of apoptosis ratio in plasma-treated cells in both cell lines. Besides, we found that the apoptosis ratio depends on cell type as well as plasma exposure time. Among two cells, H460 showed a higher intensity of Annexin V at any plasma exposure time. In addition, when we stained them with propidium iodide (PI) dye together, we found that H460 undergoes apoptosis at a faster rate than MRC5 (Supplementary Figure S4, supporting information). When many H460 cells were stained by both Annexin V and PI dyes under H2O2 stimulus, MRC5 cells were stained only by Annexin V under the same H2O2 stress. H460 cells showed higher late apoptosis rate under plasma treatment as well, which is consistent with higher cell death rate. However, we found that the lung normal cell MRC5 became smaller until 3 min plasma exposure, but became bigger in cell size after 5 min plasma treatment. As shown in the flow cytometry data in Figure 4b, both cell lines undergo time-dependent apoptotic process. It is found that the higher and faster apoptosis rate is shown in H460 cancer cells in comparison with L132 normal cells. The cell population at lower right part (Annexin V-PE-positive and 7-AAD-negative) representing the cells undergoing early apoptosis and upper right part (Annexin V-PE-positive and 7-AAD-positive) representing the cells undergoing late apoptosis of already dead cells have increased with time-dependent manner and it is shown to be much faster in H460.


Preferential killing of human lung cancer cell lines with mitochondrial dysfunction by nonthermal dielectric barrier discharge plasma.

Panngom K, Baik KY, Nam MK, Han JH, Rhim H, Choi EH - Cell Death Dis (2013)

The flow cytometry data of Annexin V and 7AAD staining of lung cells after plasma treatment. (a) The normalized cell number distribution to Annexin V intensity of H460 and MRC5 was plotted by plasma exposure time. H2O2 (1 mM) was used to make positive control. (b) Apoptosis of H460 and L132 cells was evaluated after 12 and 24 h incubation from nonthermal plasma treatment for 5 min. The number represents the percentage of cells in each quadruple
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3674375&req=5

fig4: The flow cytometry data of Annexin V and 7AAD staining of lung cells after plasma treatment. (a) The normalized cell number distribution to Annexin V intensity of H460 and MRC5 was plotted by plasma exposure time. H2O2 (1 mM) was used to make positive control. (b) Apoptosis of H460 and L132 cells was evaluated after 12 and 24 h incubation from nonthermal plasma treatment for 5 min. The number represents the percentage of cells in each quadruple
Mentions: To check whether cell death induced by plasma is apoptosis, we stained cells with Annexin V, an early apoptotic indicator. In Figure 4a, enhanced fluorescent intensity of Annexin V shows the increase of apoptosis ratio in plasma-treated cells in both cell lines. Besides, we found that the apoptosis ratio depends on cell type as well as plasma exposure time. Among two cells, H460 showed a higher intensity of Annexin V at any plasma exposure time. In addition, when we stained them with propidium iodide (PI) dye together, we found that H460 undergoes apoptosis at a faster rate than MRC5 (Supplementary Figure S4, supporting information). When many H460 cells were stained by both Annexin V and PI dyes under H2O2 stimulus, MRC5 cells were stained only by Annexin V under the same H2O2 stress. H460 cells showed higher late apoptosis rate under plasma treatment as well, which is consistent with higher cell death rate. However, we found that the lung normal cell MRC5 became smaller until 3 min plasma exposure, but became bigger in cell size after 5 min plasma treatment. As shown in the flow cytometry data in Figure 4b, both cell lines undergo time-dependent apoptotic process. It is found that the higher and faster apoptosis rate is shown in H460 cancer cells in comparison with L132 normal cells. The cell population at lower right part (Annexin V-PE-positive and 7-AAD-negative) representing the cells undergoing early apoptosis and upper right part (Annexin V-PE-positive and 7-AAD-positive) representing the cells undergoing late apoptosis of already dead cells have increased with time-dependent manner and it is shown to be much faster in H460.

Bottom Line: It is found that the cell number of lung cancer cells has been reduced more than that of the lung normal cells.The mitochondrial vulnerability to reactive species in H460 may induce distinctively selective responses.Differential mitochondrial membrane potential decrease, mitochondrial enzymatic dysfunction, and mitochondrial morphological alteration are exhibited in two cell lines.

View Article: PubMed Central - PubMed

Affiliation: Department of Plasma Bioscience and Display, Kwangwoon University, Seoul, Republic of Korea.

ABSTRACT
The distinctive cellular and mitochondrial dysfunctions of two human lung cancer cell lines (H460 and HCC1588) from two human lung normal cell lines (MRC5 and L132) have been studied by dielectric barrier discharge (DBD) plasma treatment. This cytotoxicity is exposure time-dependent, which is strongly mediated by the large amount of H2O2 and NOx in culture media generated by DBD nonthermal plasma. It is found that the cell number of lung cancer cells has been reduced more than that of the lung normal cells. The mitochondrial vulnerability to reactive species in H460 may induce distinctively selective responses. Differential mitochondrial membrane potential decrease, mitochondrial enzymatic dysfunction, and mitochondrial morphological alteration are exhibited in two cell lines. These results suggest the nonthermal plasma treatment as an efficacious modality in lung cancer therapy.

Show MeSH
Related in: MedlinePlus