研究のタイプ: 医学/生物学の研究 (experimental study)

[電力周波磁界はNB69細胞におけるフリーラジカルが関与した増殖のp38MAPK媒介性調節に影響を与える] med./bio.

Power Frequency Magnetic Fields Affect the p38 MAPK-Mediated Regulation of NB69 Cell Proliferation Implication of Free Radicals.

掲載誌: Int J Mol Sci 2016; 17 (4): 510-

この研究は、神経芽細胞株NB69の増殖への磁界MF:100 µT, 50 Hz)の影響、その影響へのp38およびc-Jun N-端末 (JNK)キナーゼの関与、さらにはMAPK-ERK1/2および-p38信号経路の活性化における活性酸素種ROS)の関与可能性を調べた。NB69培養系への100 µT MFばく露は、断続的(24、42、63時間)および連続的(15分間から120分間まで)に、それぞれp38またはJNKの阻害剤添加の有無の条件下で実施された。抗酸化物質N-アセチルシステイン(NAC)をROSスカベンジャとして用いた。その結果、MFばく露はp38、JNK、ERK1/2の一過性の活性化を誘導した;MFの増殖作用はp38阻害剤では阻害されたが、JNK阻害剤では阻害されなかった;NACは、細胞増殖およびp38活性化に対するMFの影響を阻害したが、ERK1/2活性化への影響は阻害しなかった、と報告している。

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研究目的(著者による)

The effects of exposure of neuroblastoma cells to a 50 Hz magnetic field on the cell cycle and cell proliferation and the underlying signal pathways should be investigated.

詳細情報

Previous studies by the authors (Trillo et al. 2012, Cid et al. 2012, Martinez et al. 2012, Trillo et al. 2013) have found increased cell proliferation and MAPK-ERK-1/2 signal pathway activation in human cancer cell lines after exposure to 50 Hz magnetic fields. The present study should investigate if changes in cell cycle regulation and other signaling pathways relevant to cell proliferation might be involved in the increased cell proliferation.
Cells were treated with the p38 MAPK inhibitor SB203580 (10 µM) or the JNK inhibitor SP600125 (20 µM) to investigate the involvement of these signaling pathways. Cells were also treated with N-acetyl-L-cysteine (1 mM and 10 mM) to inhibit reactive oxygen species.
The different exposure conditions and durations for individual methods were based on results from former studies.

影響評価項目

ばく露

ばく露 パラメータ
ばく露1: 50 Hz
ばく露時間: continuous for 15, 30, 60 or 120 minutes
ばく露2: 50 Hz
ばく露時間: intermittent (3 h on/3 h off) for 24, 42 or 63 hours

ばく露1

主たる特性
周波数 50 Hz
タイプ
  • magnetic field
波形
  • sinusoidal
ばく露時間 continuous for 15, 30, 60 or 120 minutes
Additional information vertically polarized
ばく露装置
ばく露の発生源/構造
チャンバの詳細 one pair of Helmholtz coils was placed inside each of two chambers magnetically shielded chambers located within two CO2 incubators
ばく露装置の詳細 in each experimental run, petri dishes (five per experimental group) were stacked in the central region of the Helmholtz coil gap, which ensured uniformity of exposure; no increase of temperature at the sample location was observed during exposure
Sham exposure A sham exposure was conducted.
Additional information the setup was composed of two identical exposure systems; following a random sequence, both coil sets and incubators were alternatively used for exposure or sham exposure
パラメータ
測定量 種別 Method Mass 備考
磁束密度 100 µT effective value 測定値 - -

ばく露2

主たる特性
周波数 50 Hz
タイプ
  • magnetic field
波形
  • sinusoidal
ばく露時間 intermittent (3 h on/3 h off) for 24, 42 or 63 hours
Additional information vertically polarized
ばく露装置
ばく露の発生源/構造
  • E1と同じ装置
Sham exposure A sham exposure was conducted.
パラメータ
測定量 種別 Method Mass 備考
磁束密度 100 µT effective value 測定値 - -

Reference articles

ばく露を受けた生物:

方法 影響評価項目/測定パラメータ/方法

研究対象とした生物試料:
調査の時期:
  • ばく露中
  • ばく露後

研究の主なアウトカム(著者による)

The cell cycle distribution showed significant changes after 24 h (increased proportion of cells in S phase) and 42 h (decreased proportion of cells in G0 phase/G1 phase) of exposure compared to the control group. Exposure to the magnetic field for 120 minutes significantly increased the protein expression of cyclin D1 and decreased that of p27Kip1.
As expected, cells exposed to the magnetic field for 63 hours showed a significantly increased cell proliferation. The proliferative effect of the magnetic field was blocked by the p38 MAPK inhibitor, but not by the JNK inhibitor. Moreover, the proliferative effect of the MF was blocked by 1 mM of N-acetyl-L-cysteine while a concentration of 10 mM significantly reduced cell proliferation compared to the control group with and without co-exposure to the magnetic field.
After 15 minutes and 30 minutes of exposure, the protein expression of phosphorylated p38 MAPK was significantly increased compared to the control group and this increase could be blocked by treatment with the p38 MAPK inhibitor or N-acetyl-L-cysteine. The protein expression of phosphorylated JNK was significantly increased after 15 min and significantly decreased after 60 min of exposure compared to the control group. The phosphorylated ERK-1/2 protein expression was significantly increased after 30 minutes of exposure and was not influenced by N-acetyl-L-cysteine treatment.
The authors conclude that exposure of neuroblastoma cells to a 50 Hz magnetic field might induce changes in cell cycle and increase cell proliferation. The effects are probably mediated by a radical-dependent activation of the p38 MAPK and a radical-independent activation of the ERK-1/2 signal pathways.

研究の種別:

研究助成

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