大气压冷等离子体射流诱导乳癌细胞凋亡研究

STUDY ON APOPTOSIS OF BREAST CANCER CELLS INDUCED BY COLD ATMOSPHERIC-PRESSURE PLASMA JET

目的研究大气压冷等离子体射流对乳癌细胞系MDA-MB-468细胞的凋亡诱导作用。方法采用自行研制的大气压冷等离子体射流发生装置,处理MDA-MB-468细胞。大气压冷等离子体射流处理时间分别为10、30、60和90s,以不处理者为空白对照组,以单纯气流处理90s者为实验对照组。采用四甲基偶氮唑盐(MTT)法检测各组细胞增殖抑制率,二脒基苯基吲哚(DAPI)染色检测各组细胞的核形态变化,荧光定量聚合酶链反应(FQ-PCR)测定各组细胞Bax和Bcl-2mRNA的表达变化。结果大气压冷等离子体射流可明显抑制细胞的增殖,且有明显的剂量依赖性(F=22.4、42.3,P<0.01),而单纯气流对MDA-MB-468细胞无明显影响。经大气压冷等离子体射流处理后细胞出现核固缩及核碎裂。大气压冷等离子体射流作用后细胞Bax的表达上调,Bcl-2的表达下调(F=32.3、12.4,P<0.05)。结论大气压冷等离子体射流对MDA-MB-468细胞有增殖抑制作用,可能与Bax表达上调及Bcl-2表达下调诱导细胞凋亡有关。

Objective To study the effect of cold atmospheric-pressure plasma jet （CAPPJ） on apoptosis of MDA-MB- 468 cells in breast cancer cell lines. Methods A self made CAPPJ device was applied in this study for MDA-MB-468 cells. The cells were exposed to CAPPJ for 10,30 and 90 seconds, respectively. That with no treatment served as hlank control group, that exposed to air stream only for 90 seconds served as experimental control group. By using methyl thiazolyl tetrazolium （MTT） assay, the cell proliferation inhibiting rate in each group was determined; by using DAPI dyeing, the morphologic changes of cell nucleus in each group were detected. The mRNA expressions of Bax and Bcl-2 were determined by real-time quantitative polymerase chain reaction （Real-time PCR）. Results CAPPJ could obviously inhibit the cell proliferation with dose-dependent in nature （F=22.4,42.3 ;P〈0.01）, and air stream only did not impact MDA-MB-468 cells. After exposing to CAPPJ, pyknosis and nuclear fragmentation occurred, Bax expression up regulated, and Bcl-2 down-regulated （F=32.3,12.4;P〈0.05）. Conclusion The CAPPJ can inhibit the growth of MDA-MB-468 cells through inducing apoptosis. The mechanism may be associated with promotion of Bax and inhibition of Bcl 2 gene.