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目的探讨维生素B6(VB6)缺乏对人类BRCA1突变乳腺癌淋巴细胞株基因组稳定性的影响。方法以VB6血浆正常生理浓度为依据(~20 nmol/L),设置0,6,12,24,48,96,200 nmol/L 7个浓度组,常规RPMI1640培养基为对照(VB6浓度4800 nmol/L),培养携带BRCA1突变的乳腺癌患者淋巴细胞株(GM13705)9 d,利用细胞质阻断微核分析(CBMN),评价VB6缺乏对该细胞株细胞活性及遗传稳定性的影响。结果在0 nmol/L VB6浓度下,细胞不能存活;6,12 nmol/L活细胞数有下降趋势,24 nmol/L时开始增长,与初次培养时无显著性差异,48 nmol/L较24 nmol/L显著上升(P<0.05);在96时达最大值,与200,4800 nmol/L组间无显著性差异。双核细胞微核(MNBN)频率随VB6浓度增加显著下降,48 nmol/L时降至最低(P<0.001～0.01);48,96,200,4800 nmol/L时4个浓度间无显著性差异。结论 VB6浓度在96 nmol/L时是受试细胞株活性最佳浓度,48 nmol/L可维持受试细胞株遗传稳定性的最佳状态。
Objective To investigate the effect of vitamin B6 (VB6) deficiency on the genomic stability of human breast cancer cell line BRCA1. Methods Seven concentrations of 0, 6, 12, 24, 48, 96 and 200 nmol / L were set up based on normal physiological concentrations of VB6 plasma (~ 20 nmol / L). The conventional RPMI1640 medium was used as a control (VB6 concentration 4800 nmol / L ), And cultured BRCA1-bearing breast cancer cell line (GM13705) for 9 days. The effects of VB6 deficiency on the cell viability and genetic stability of the cell line were evaluated by cytoplasmic blocking micronucleus assay (CBMN). Results At 0 nmol / L VB6 concentration, the cells could not survive. The number of viable cells at 6 and 12 nmol / L showed a decreasing trend and started to increase at 24 nmol / L with no significant difference from that at 48 nmol / L nmol / L increased significantly (P <0.05). At 96 hours, there was no significant difference between 200 and 4800 nmol / L groups. The frequencies of MNBN decreased significantly with the increase of VB6 concentration and decreased to the lowest at 48 nmol / L (P <0.001 ~ 0.01). There was no significant difference between 48, 96, 200 and 4800 nmol / L concentrations. Conclusion VB6 concentration of 96 nmol / L is the best concentration of test cell activity, 48 nmol / L can maintain the genetic stability of the best test cell strains.