摘要
研究硼中子俘获疗法（BNCT）体外杀伤人黑色素瘤细胞的效应及机制。方法 首先检测黑色素瘤细胞A375吸收含硼化合物二羟基苯丙氨酸硼（BPA）的情况,然后采用医院中子照射器（IHNI-1）对含硼（10B）细胞进行照射。克隆存活实验检测细胞的放射敏感性,MTT法检测细胞增殖率,流式细胞术检测凋亡,Western blot检测胞质内细胞色素C表达和caspase-9的激活。结果 BPA孵育24 h,A375细胞10B浓度为（2.884±0.148）μg/107个细胞,达到了BNCT杀伤细胞的要求。富含10B的细胞经中子照射2.1 min后存活分数降低为对照组的58%（t=2.964,P〈0.05）,细胞经中子照射后24 h增殖率下降为对照组的83%（t=3.286,P〈0.05）,BNCT组细胞凋亡率达（55.2±7.9）%,明显高于对照组（t=9.754,P〈0.05）,胞质内细胞色素C水平上升且caspase-9激活程度增加（t=7.625、8.307,P〈0.05）。结论 BNCT能够杀伤黑色素瘤细胞,其机制可能通过线粒体途径诱导细胞凋亡。
Objective To study the effect and underlying mechanism of boron neutron capture therapy （BNCT） on human melanoma cells. Methods The situation of boronophenylalanine （BPA） uptake of human melanoma cells A375 was detected and then the boron-10 （10B） enriched cells were irradiated by an in-hospital neutron irradiator （IHNI-1）. The radiation sensitivity was measured using clonogenic survival assay, the proliferation was examined by MTT assay, apoptosis was determined using flow cytometry, and the protein expression of cytochrome C in cytosol and activation of caspase-9 was detected by Western blot.Results 10B concentration in A375 cells approached to （2.884±0.148） μg/107 cells after 24 h culture with BPA, which met the requirement of BNCT. At 2.1 min after neutron radiation, the survival fraction of BNCT group was decreased to 58% of control （t=2.964,P〈0.05）. At 24 h after BNCT, the cell viability was decreased to 83% of control （t=3.286, P〈0.05）, the apoptosis ratio was （55.2±7.9）%（t=9.754, P〈0.05）, and the protein level of cytochrome C in cytosol and the activativity of caspase-9 were also enhanced （t=7.625,8.307, P〈0.05）.Conclusions Human melanoma cells can be killed by BNCT due to apoptosis through a mitochondrial pathway.
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2013年第1期10-13,共4页
Chinese Journal of Radiological Medicine and Protection
基金
国家自然科学基金（81207142）
江苏省医学领军人才与创新团队项目（K201106）
关键词
硼中子俘获疗法
二羟基苯丙氨酸硼
医院中子照射器
凋亡
Boron neutron capture therapy
Boronophenylalanine
In-hospital neutronirradiator
Apoptosis
