CAJ | 학술논문

目的研究硼中子俘获疗法(BNCT)对体外培养人胶质瘤干细胞(GSCs)周期进程的影响及机制,比较GSCs和胶质瘤细胞株SHG-44对BNCT敏感性的差异.方法首先检测人GSCsSU2和人胶质瘤细胞株SHG-44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,流式细胞术检测细胞周期进程,Westem blot检测cyclin B1、CDK1和P21[WAF1]蛋白表达情况.结果 5 mmol/L BPA孵育SU2和SHG-44细胞24h,10B浓度分别可达(1.76±0.28)和(2.50±0.12)μg/107细胞,富含10B的细胞经IHNI-1照射后生存率和增殖率与不含10B的细胞比均明显下降(P＜ 0.05或P＜0.01),SU2细胞经IHNI-1照射4 min时BNCT敏感性低于SHG-44细胞(P＜0.05).与未加BPA而仅用中子照射组比较,经BNCT后G2/M期SU2和SHG-44细胞比例增高(P＜0.05),cyclin B1和CDK1蛋白表达降低(P＜0.01),P21[WAF1蛋白表达增加(P＜0.01).结论 GSCs的BNCT敏感性低于胶质瘤细胞,BNCT可通过抑制细胞周期蛋白形成将细胞阻滞于G2/M期来抑制细胞更新和增殖.
목적 연구붕중자부획요법(BNCT)대체외배양인효질류간세포(GSCs)주기진정적영향급궤제,비교GSCs화효질류세포주SHG-44대BNCT민감성적차이.방법 수선검측인GSCsSU2화인효질류세포주SHG-44흡수함붕화합물이간기분병안산붕(BPA)적정황,연후채용의원중자조사기(IHNI-1)대함붕(10B)세포진행조사,극륭존활실험검측세포적방사민감성,MTT법검측세포증식솔,류식세포술검측세포주기진정,Westem blot검측cyclin B1、CDK1화P21[WAF1]단백표체정황.결과 5 mmol/L BPA부육SU2화SHG-44세포24h,10B농도분별가체(1.76±0.28)화(2.50±0.12)μg/107세포,부함10B적세포경IHNI-1조사후생존솔화증식솔여불함10B적세포비균명현하강(P＜ 0.05혹P＜0.01),SU2세포경IHNI-1조사4 min시BNCT민감성저우SHG-44세포(P＜0.05).여미가BPA이부용중자조사조비교,경BNCT후G2/M기SU2화SHG-44세포비례증고(P＜0.05),cyclin B1화CDK1단백표체강저(P＜0.01),P21[WAF1단백표체증가(P＜0.01).결론 GSCs적BNCT민감성저우효질류세포,BNCT가통과억제세포주기단백형성장세포조체우G2/M기래억제세포경신화증식.
Objective To study the effect and mechanism of boron neutron capture therapy (BNCT) on cell cycle procession in human glioma stem cells (GSCs) in vitro,and to analyze the difference of sensitivity for BNCT on GSCs and glioma cells.Methods Firstly,uptake of boronophenylalarine (BPA) on human GSCs SU2 and glioma cells SHG-44 was detected.Then the cells enriched boron-10 (10B) were irradiated by In-Hospital Neutron Irradiator (IHNI-1).The radiation sensitivity was measured using clonogenic survival assay.The proliferation was examined by MTT assay.The cell cycle procession was determined using flow cytometry.The protein expression of cyclin B1,CDK1 and P21[WAF1] were detected by Western blot.Results 10B concentration of SU2 and SHG-44 cells arrived at (1.76 ±0.28) and (2.50 ±0.12) μg/107cells at 24 h in 5 mMBPA.After radiation by IHNI-1,the survival fraction and viability of cells enriched 10B were decreased significantly (P ＜ 0.05 orP ＜ 0.01),compared with those of 10B-free.The BNCT sensitivity of SU2 cells was lower than that of SHG-44 cells (P ＜ 0.05).The proportion of G2/M phase of SU2 and SHG-44 cells was increased after BNCT compared with that of 10B-free(P ＜ 0.05).The protein levels of cyclin B1 and CDK1 were decreased(P ＜ 0.01),and that of P21[WAF1] was increased(P ＜ 0.01).Conclusions The BNCT sensitivity of GSCs was lower than that of glioma cells.BNCT could inhibite cell regeneration and proliferation and make G2/M to be blocked by inhibition of cell cycle protein formation.