肝素干预长春新碱诱导的K562细胞凋亡

目的探讨肝素对长春新碱诱导的K562细胞凋亡及增生的影响。方法肝素预处理K562细胞1h，再经长春新碱（0．05mg／L）处理24h，应用Hoechst33342染色、DNA电泳、FMC等方法检测细胞凋亡。Trypanblue染色及MTr法检测细胞毒性及增生反应。结果Hoechst33342荧光染色见长春新碱凋亡诱导组细胞凋亡率达40．10％，肝素25、50、100、200U／ml组凋亡率依次为32．47％、29．70％、25．50％、19．53％（均P〈0．05）。随肝素浓度增加DNA电泳凋亡梯带亮度逐渐减弱至消失。FACS检测凋亡诱导组凋亡率为21．61％，肝素25～200U／ml组凋亡率依次为13．64％、11．75％、8．59％、6．03％（均P〈0．05）。肝素各组处理K562细胞24h后细胞存活率、活细胞总数及细胞增生水平与正常对照组比较差异均有统计学意义（P〉0．05）。结论肝素浓度在200U／ml内对K562细胞无毒性作用及增生影响，在25～200U／ml以浓度依赖性方式抑制由长春新碱诱导的K562细胞凋亡。

The influence of heparin on the apoptosis and proliferation for K562 cells induced by vincristine

Objective To investigate the effect of heparin on the apoptosis and proliferation of human myeloid leukemia cell line K562 induced by vincristine.Methods K562 cells were pretreated by heparin for 1h,then cultured with 0.05 mg/L vincristine in 37℃ 5% CO2 for 24 h.Apoptosis of KS62 cells was evaluated by Hoechst 33342 staining,flow cytometer and DNA agarose gel electrophoresis after culture for 24 hours.The effect of heparin on KS62 cell proliferation and toxicitv was determined by Trypan blue staining and MTT assay.Results In the apeptosis induced group,the apeptosis rate was 40.10% dected by Hoechst 33342 fluorescence staining.The hepafin in different concentrations was found to be able to inhibit the apoptosis of K562 cells triggered by vincristine and the apoptosis rate was 32.47%,29.7%,25.5%,19.53% in the heparin groups of 25,50,100,200 U/ml,respectively.The apeptosis rate was significantly lower in the apeptosis induced group than in the heparin groups of 25,50,100,200 U/ml(P＜0.05).The typical DNA ladder could be found in the apoptosis-induced group,and the DNA ladder gradually disappeared along with the increase of heparin(5～200 U/ml).The sub-G1 peak of K562 cells could be found in the induced group by FACS and the apoptosis rate was 21.61%.In the heparin groups of 25,50,100,200 U/ml,the sub-G1 Peak of K562 cells gradually dropped and the apoptosis rate was 13.64%,11.75%,8.59%,6.03%(P＜0.05),respectively.After K562 cells were incubated with different hepafin concentrations(5～200 U/ml)for 24 hours,there was no difference compared with the normal control group in both the total live cell numbers and the cell proliferation rate measured by trypan blue staining and MTT assay(P＞0.05).Conclusion The results suggested that heparin had no influence on KS62 cell toxicity and proliferation,but may inhibit the apoptosis of KS62 cells induced by vincristine.