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Cold atmospheric-pressure nitrogen plasma induces the production of reactive nitrogen species and cell death by increasing intracellular calcium in HEK293T cells

Author:
Iuchi, Katsuya, Morisada, Yukina, Yoshino, Yuri, Himuro, Takahiro, Saito, Yoji, Murakami, Tomoyuki, Hisatomi, Hisashi
Source:
Archives of biochemistry and biophysics 2018 v.654 pp. 136-145
ISSN:
0003-9861
Subject:
atmospheric pressure, calcium, cell death, cell proliferation, chelating agents, culture media, human cell lines, humans, hydrogen peroxide, irradiation, neoplasms, nitrates, nitric oxide, nitrites, nitrogen, reactive nitrogen species, therapeutics, viability
Abstract:
Cold atmospheric-pressure plasma (CAP) has been emerging as a promising tool for cancer therapy in recent times. In this study, we used a CAP device with nitrogen gas (N2CAP) and investigated the effect of the N2CAP on the viability of cultured cells. Moreover, we investigated whether N2CAP-produced hydrogen peroxide (H2O2) in the medium is involved in N2CAP-induced cell death. Here, we found that the N2CAP irradiation inhibited cell proliferation in the human embryonic kidney cell line HEK293T and that the N2CAP induced cell death in an irradiation time- and distance-dependent manner. Furthermore, the N2CAP and H2O2 increased intracellular calcium levels and induced caspase-3/7 activation in HEK293T cells. The N2CAP irradiation induced a time-dependent production of H2O2 and nitrite/nitrate in PBS or culture medium. However, the amount of H2O2 in the solution after N2CAP irradiation was too low to induce cell death. Interestingly, carboxy-PTIO, a nitric oxide scavenger, or BAPTA-AM, a cell-permeable calcium chelator, inhibited N2CAP-induced morphological change and cell death. These results suggest that the production of reactive nitrogen species and the increase in intracellular calcium were involved in the N2CAP-induced cell death.
Agid:
6093061