摘要
目的:观察一氧化氮对肿瘤细胞SMMC-7721辐射敏感性的作用效果。方法:实验于2005-06/09在兰州大学生命科学学院和中科院近代物理研究辐射医学实验室进行。处于对数生长期的肝癌细胞SMMC-7721,在用X射线照射前4h,换入含有0.1mmol/L硝普钠(一氧化氮的前体)的培养液,与对照组(不加硝普钠)一起,在200cGy/min的剂量率下,分别照射0,1,2,4,6,8Gy,换为正常培养液培养。用集落形成法计算细胞的存活率,用吖啶橙/溴乙啶双染法检测细胞的死亡情况,用流式细胞仪检测细胞周期。结果:①存活曲线细胞存活率随照射剂量增加而减少,硝普钠组细胞的克隆形成率低于对照组(2Gy时,P<0.01)。②细胞死亡百分率(坏死细胞与凋亡细胞总数/总细胞数):与照射剂量呈正相关,硝普钠组高于对照组(P<0.05)。对照组从(9.95±3.53)%(0Gy)逐渐升至(58.74±3.46)%(6Gy),而硝普钠组则从(18.53±12.02)%(0Gy)迅速升至(61.57±9.53)%(2Gy)。③细胞周期检测结果:对照组细胞经过X射线照射后,出现了G2/M期阻滞[从0Gy时(12.50±5.76)%逐渐增加到8Gy(40.36±2.74)%],而硝普钠组细胞在低剂量时主要表现为G0/G1期阻滞[0Gy:(16.06±7.19)%;2Gy:(17.93±0.92)%],而G2/M期阻滞仅在高剂量时明显[8Gy时为(50.10±3.93)%,P<0.05]。结论:经硝普钠产生的一氧化氮,通过与X射线协同作用,减少了肝癌细胞SMMC-7721的细胞存活率,促进细胞死亡,阻止细胞被阻滞至G2/M期,是一种有效的辐射增敏剂。
AIM： To investigate the effect of nitnc oxide （NO） on the radiosensitivity of the tumor cell SMMC-7721.
METHODS： The experiment was carried out in the School of Life Sciences, Lanzhou University and Laboratory of Radiation Medicine, Institute of Modem Physics in the Chinese Academy of Sciences between June and September 2005. SMMC-7721 cells at exponential phase of growth ware pre-incubated with 0.1 mmol/L sodium nitroprusside （SNP, a NO releaser） for 4 hours, and then irradiated by X-ray at 0, 1, 2, 4, 6 or 8 Gy with dose rate of 200 cGy/min. The cell survival fractions ware measured by colony-formation assay, the cell apoptosis was detected by acridine orange/ethylene dibromide staining, and the cell cycle ware obtained with flow cytometry.
RESULTS： ①The survive curve： The survival rate had a negative correlation with the irradiation dose, and the cloning efficiency of the cells treated with SNP was much lower than that of controls （2 Gy, P 〈 0.01）.②The percentages of the apoptosis cells （total number of necrosis cells and apoptosis cells/total number of cells） had a positive correlation with the irradiation dose, and ware significantly higher in the groups pre-incubated with SNP than in control group （P 〈 0.05） [control group： （9.95±3.53）% （0 Gy） to （58.74±3.46）% （6 Gy）; SNP group： （18.53±12.02）% （0 Gy） to （61.57±9.53）% （2 Gy）].③Cell cycle： For SMMC-7721 cells, G2/M accumulation was prominently induced by X-ray [0 Gy （12.50±5.76）% to 8 Gy （40.36±2.74）%], but the groups pre-incubated with SNP ware significant in Go/G1 accumulation [0 Gy： （16.06± 7.19）%; 2 Gy： （17.93±0.92）%], and G2/M accumulation was significant at higher dose of irradiation [8 Gy： （50.10±3.93）%, P 〈 0.05].
CONCLUSSION： With the synergism of X-ray, SNP-induced NO can decrease the survival rate of SMMC-7721 cells, improve the cell apoptosis, and prevent the G2/M accumulation, so as to be effective in the treatment of radiotherapy.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第8期1477-1480,I0003,共5页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
中国科学院2002年度"百人计划"基金项目(0306010BR0)
甘肃省科技攻关项(2GS052-A43-008-02)~~
