摘要
目的观察经苯代谢物长期处理的K562细胞在撤出苯代谢物后,继续传代培养时的红系分化能力的变化规律。方法K562细胞分别以5μmol/L 1,2,4-苯三醇和10μmol/L氢醌处理4周,每周以联苯胺染色法检测受处理细胞Hemin诱导的红系分化率,评价红系分化能力;处理满4周后,撤出苯代谢物,一部分细胞继续传代培养,持续12周,每2周测定细胞的红系分化能力,另一部分细胞进行单细胞克隆培养,选取红系分化能力较低的细胞株连续传代培养,每2周测定各细胞株红系分化能力。结果在处理的前3周,苯三醇组和氢醌组K562细胞红系分化能力出现时间依赖性下降,而在处理3和4周后,苯三醇组红系分化率均分别只有对照组的51.1%和50.1%,氢醌组均只有对照组的33.1%和31.4%。4周处理后,洗去苯代谢物,在继续传代培养的前6周,苯三醇组和氢醌组红系分化率均分别稳定地维持在22.3%~23.9%和14.0%~15.6%范围内,此后逐渐回升,但仍然明显低于对照组。经苯代谢物处理4周后再经4周的单细胞克隆培养,获得单细胞起源克隆细胞株,各组来源的细胞克隆的红系分化率分别为:对照组39.1%~49.7%,平均45.7%;苯三醇组4.1%~45.3%,平均20.6%;氢醌组0.8%~28.9%,平均12.7%。如将红系分化率≤10%的单克隆细胞株进行第1轮继续传代培养2周,苯三醇组和氢醌组来源细胞株的平均红系分化率仅有小幅回升。如每处理组每批次各挑选出3个红系分化能力最低的单克隆细胞株,再进行第2轮继续传代培养2周,则单克隆细胞株在刚克隆出来时、第1次和第2次继续传代培养后的平均红系分化率,在苯三醇组来源细胞株中分别为7.5%、8.4%和16.3%,在氢醌组来源细胞株中分别为2.3%、3.4%和7.5%。从每处理组的每批次克隆细胞株中选取红系分化能力最低的细胞株,进行4轮连续传代培养,经最初4周的单细胞克隆培养后、第1次、第2次、第3次和第4次2周继续传�
Objective To obseve the changes of erythral differentiation abilities of K562 cells treated with benzene metabolites for long-term after benzene metabolites were withdrawn and the cells continuously cultured in the forms of cell mixture or single cell cloning.Methods Firstly,K562 cells were treated with 1,2,4-benzenetriol and hydroquinone at the final concentrations of 5μmol/L and 10μmol/L for four weeks,respectively.The hemin-induced erythroid differentiation rate of treated cells was detected by benzidine staining weekly,to evaluate the erythroid differentiation ability.After treatment with benzene metabolites for four weeks,benzene metabolites were withdrawn,one part of the cells continued to subculture for 12 weeks,and the erythroid differentiation abilities of the cells were determined once every two weeks;the rest another part of the cells were cultured in single cell cloning to obtain the single cell-derived strains,then the cell strais with lower erythroid differentiation ability were selected to continuous subculture,and the erythroid differentiation abilities of the cell strains were determined once every two weeks.Results In the first three weeks of treatment,the erythroid differentiation abilities in the benzenetriol group and the hydroquinone group decreased in a time-dependent manner.After three and four weeks,the hemin-induced erythroid differentiation rates in the benzenetriol group were only 51.1%and 50.1%of that in the control group,and the hemin-induced erythroid differentiation rates in the hydroquinone group were only 33.1%and 31.4%of that in the control group.After four weeks of treatment,the benzene metabolites were removed,and the cells continuously subcultured.The hemin-induced erythroid differentiation rates in the benzenetriol group and hydroquinone group were stable during the first six weeks of subculture;after that,the hemin-induced erythroid differentiation rates increased gradually,but being still significantly lower than that in the control group.After four weeks of treatment wi
作者
刘小凡
余春红
巩萌
张宇婧
韩畅
易宗春
LIU Xiao-fan;YU Chun-hong;GONG Meng;ZHANG Yu-jing;HAN Chang;YI Zong-chun(School of Biological Science and Medical Engineering,Beijing Advanced Innovation Center for Biomedical Engineering,Beihang University,Beijing 100083,China;School of Medical Science and Engineering,Beihang University,Beijing 100083,China)
出处
《毒理学杂志》
CAS
CSCD
2022年第1期49-54,共6页
Journal of Toxicology
基金
国家自然科学基金(81573192,81072325)。
关键词
苯代谢物
1
2
4-苯三醇
红系分化
氢醌
Benzene metabolites
1,2,4-Benzenetriol
Erythroid differentiation
Hydroquinone
