目的 研究红花注射液对白血病HEL 细胞增殖、凋亡的影响,并初步探讨其相关分子机制。方法 HEL 细胞随机用或不用红花注射液处理,不用红花注射液处理者为对照组。不同浓度(10、20、30、40、50 mg/mL)的红花注射液作用于HEL 细胞24、48、72 h 后,采用四甲基偶氮唑蓝(MTT)法检测各组细胞增殖能力;10、20、30 mg/mL 红花注射液处理HEL 细胞48 h 后,采用流式细胞术检测各组细胞细胞周期与凋亡的变化,RT-PCR 法检测细胞HOXB3 mRNA 的表达。结果 HEL 细胞经浓度为10、20、30、40、50 mg/mL的红花注射液处理24、48 及72 h 后,与对照组比较,细胞增殖均不同程度地受抑制,且随着红花注射液浓度的增加,细胞增殖抑制率增加(P<0.05)。不同浓度红花注射液作用HEL 细胞48 h 后,细胞周期的分布发生改变,与对照组相比G2/M 期细胞数显著增多,S 期细胞数显著减少(P<0.05),且呈剂量依赖性。随着红花注射液浓度的增加,细胞凋亡率增加,以浓度依赖的方式诱导细胞凋亡(P<0.05)。与对照组比较,各浓度红花注射液组HEL 细胞中HOXB3 mRNA 表达均下降(P<0.05)。结论 红花注射液体外可有效抑制HEL 细胞的增殖并诱导其凋亡,其相关分子机制可能与下调HOXB3基因的表达有关。
Abstract
Objective To study the effect of safflower injection on the proliferation and apoptosis of human leukemia cell line HEL and the relevant molecular mechanisms.Methods HEL cells were treated with different concentrations of safflower injection. HEL cells without safflower injection treatment were used as the control group. MTT method was used to detect the inhibitory rate of the HEL cells at 24, 48 and 72 hours after various concentrations of safflower injection treatment (10, 20, 30, 40 and 50 mg/mL). The cell cycle and apoptosis were detected using flow cytometry and the HOXB3-mRNA expression was measured by RT-PCR at 48 hours after safflower injection treatment (10, 20 and 30 mg/mL).Results Compared with the control group, various concentrations of safflower injection inhibited HEL cell proliferation in a dose-dependent manner (P<0.05). At 48 hours after various concentrations of safflower injection treatment, the number of treated cells in the G2/M phase increased, but that in the S phase decreased, and the apoptosis rate was significantly higher than that in the control group, with a dose-dependent manner (P<0.05). The expression of HOXB3-mRNA in safflower injection-treated cells decreased in a dose-dependent manner compared with the control group (P<0.05).Conclusions Safflower injection can inhibit proliferation and induce apoptosis of HEL cells in vitro, and its underlying mechanisms may involve down-regulation of the HOXB3-mRNA expression.
关键词
红花注射液 /
白血病 /
细胞增殖 /
细胞凋亡 /
HOXB3 /
人类白血病细胞株
Key words
Safflower injection /
Leukemia /
Cell proliferation /
Cell apoptosis /
HOXB3 /
Human leukemia cell line
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基金
山东省自然科学基金项目(ZR2014HL032);山东省医药卫生科技发展计划项目(2014WS0183);滨州市科技发展计划项目(2014ZC0113);滨州医学院科技计划项目(BY2013KJ03)