摘要
目的观察微波辐射对人成神经母细胞瘤SH-SY5Y细胞增殖与分化的影响。方法采用10、30、50 mW/cm2辐射强度的微波辐射SH-SY5Y细胞5 min,0mW/cm2为对照组,锥虫蓝拒染法进行细胞计数并计算增殖率;20μmol/L全反式维甲酸黑暗下诱导分化72 h后,在光学显微镜下观察细胞分化的形态并计算分化细胞率;流式细胞仪检测细胞周期;蛋白质印迹检测细胞神经纤丝重链亚基的蛋白表达。结果微波辐射后随辐射强度增加,细胞增殖率在各时间点上均出现不同程度的梯度抑制,与对照组(0mW/cm2)相比,10 mW/cm2辐射组在72 h时出现明显抑制;30 mW/cm2辐射组在48 h时出现明显抑制;50 mW/cm2辐射组在辐射12 h时就出现明显抑制;20μmol/L全反式维甲酸黑暗下诱导分化72 h后,30、50 mW/cm2微波辐射组细胞分化的数量少于对照组细胞,而且细胞轴突长度短于对照组细胞;随着辐射强度的增加,G0/G1期的SH-SY5Y细胞比例逐渐增加,S期细胞比例逐渐减少,G2+M期细胞比例变化相对较小;微波辐射后,SH-SY5Y细胞总NF-H蛋白表达无明显变化,但磷酸化的NF-H蛋白表达随辐射强度的增加逐渐降低。结论微波辐射可同时抑制SH-SY5Y细胞增殖与分化并呈现明显的剂量依赖性抑制作用;细胞周期呈G0/G1期阻滞;对NF-H的表达无影响,但明显抑制了NF-H的磷酸化。
Objective The influence of microwave radiation in the proliferation and differentiation of human neuroblastoma SH-SY5Y cells were investigated.Methods The SH-SY5Y cells were radiated for 5 minutes by 0（the control group）,10,30,50 mW/cm2 of microwave.The prolife ration rates were counted by trypan blue staining.After the cells were induced by 20 μmol/L all-trans retinoic acid（ATRA） in the dark 72 h,the differentiation rates were calculated under the light microscope.The cell cycles were analyzed by flow cytometry（FCM）.The expressions of NF-H proteins were detected by Western blotting.Results The proliferation rates of SH-SY5Y cells were reduced gradually with the increases of microwave radiation intensity.Compared with control group,the proliferation of SH-SY5Y cells were significantly inhibited in 10,30,50 mW/cm2 radiation group respectively at 72,48 and 12 h after microwave radiation.After the cells were induced 72 h by 20 μmol/L ATRA,the numbers of differentiated cells of 30,50 mW/cm2 radiation groups were less than control group,and the cell axons of various radiation groups were shorter than control group.With the increase of radiation intensity,microwave increased the proportion of SH-SY5Y cells in G0/G1 phase,and decreased the proportion in the S phase.The proportion of SH-SY5Y cells in G2＋M phase cells was unchanged.After microwave radiation,the expressions of NF-H protein were not changed,but the expressions of phosphorylated NF-H proteins were decreased gradually.Conclusion The proliferation and differentiation of SH-SY5Y cells can be inhibited by microwave radiation in an obvious dose-dependent manner.Cell cycle progression was blocked in the G0/G1 phase.Microwave had no effect on the expression of NF-H in SH-SY5Y cells,but could inhibit NF-H phosphorylation significantly.
出处
《肿瘤防治研究》
CAS
CSCD
北大核心
2012年第6期671-675,共5页
Cancer Research on Prevention and Treatment
基金
吉林省科技厅白求恩基金资助课题(200705408)
吉林省教育厅科研基金资助课题(2009315)
吉林医药学院科研基金资助课题(200903)
