低浓度力达霉素对人白血病细胞K562增殖的抑制作用

Inhibitory effect of low concentration of LDM on the proliferation of human leukemia K562 cells

目的探讨低浓度力达霉素(LDM)对人白血病细胞K562增殖的抑制作用及其可能的作用机制。方法以不同浓度LDM处理K562细胞48 h,通过MTT法检测LDM对细胞增殖的抑制作用;利用苔盼蓝染色对细胞活性进行测定;利用瑞氏-姬姆萨染色法观察LDM对细胞形态的影响;Elisa法检测肿瘤坏死因子-α(TNF-α)在细胞培养上清中的表达;采用Western Blot检测凋亡相关蛋白TNF-α的表达。结果 LDM对K562细胞增殖有一定的抑制作用,用10-12、10-11、10-10、10-9、10-8mol/L浓度的LDM分别作用K562细胞48 h,抑制率分别为(21.2±2.48)%、(37.5±0.88)%、(52.1±1.24)%、(64.5±1.61)%和(71.3±2.00)%。通过瑞氏姬姆萨染色,荧光显微镜下可见胞膜皱缩,有泡状突起及不规则凹陷,胞核染色质浓缩,固缩,浓染,断裂成团块分散在胞膜周边。酶联免疫吸附试验(ELISA)检测细胞培养上清,LDM处理组与对照组相比,TNF-α表达水平升高。Western Blot分析显示,分别用10-8、10-10、10-12 mol/L浓度的LDM处理细胞48 h后,TNF-α蛋白的表达量依次为0.42±0.01、1.67±0.13(P<0.01)和2.41±0.08(P<0.01),与对照组0.23±0.02相比,吸光度值比升高。结论低浓度LDM可能通过上调TNF-α表达水平诱导K562细胞凋亡抑制其增殖。

[Objective]To explore the inhibitory effect of low concentrations of lidamycin （LDM） on human leukemia K562 cells proliferation and its possible mechanism. [Methods]With different concentrations of LDM for 48 h treatment of K562 ceils, inhibitory effect of LDM on cell proliferation was observed through the detection of MTr. Trypan blue staining was used to detect cell activity. Influences on morphologic changes of cells were examined with Wright-Giemsa coloring analysis. Expression of TNF- in K562 cells was measured with Western Blot.[Results]LDM has a certain inhibitory effect on the proliferation of K562 cells. With 10-12 tool/L, 10-11 mol/L, 10-10 mol/L, 10-9 mol/L, 10-8 mol/L LDM on K562 cells for 48 h, the inhibitory rates were （21.2±2.48）%, （37.5±0.88）%, （52.1±1.24）%, （64.5±1.61）% and （71.3±2.00）%. By Wright-Giemsa staining, membrane blebbing, vesicular apophysis and irregular depressions, cytoplasmic and nucleus chromiatin condensation, concentration, breaking into clumps scattered around the cell membrane were observed under fluorescence microscope. ELISA was used to detect cell culture supematant, Compared LDM treatment group with the control group, express level of TNF-α was elevated. Western Blot analysis showed, by using 10-8 mol/L, 10-10 tool/L, 10-12 mol/L LDM treating cells for 48 h, the expression of TNF-α protein were 0.42±0.01, 1.67±0.13 （P〈0.01） and 2.41±0.08 （P〈0.01）, compared with 0.23±0.02 of the control group, the absorbance ratio was increased. [Conclusion]LDM of low concentration can inhibit the proliferation and induce apoptosis of K562 cells via up-regulating the expression of TNF-a.