青蒿素衍生物对宫颈癌HeLa细胞体外增殖及凋亡能力的影响及其分子机制研究

Analysis of the effects of artemisinin derivatives on proliferation and apoptosis of HeLa cells in cervical cancer and its molecular mechanism

目的:观察青蒿素衍生物(双氢青蒿素和青蒿琥酯)对宫颈癌细胞体外增殖及凋亡能力的影响,并探讨其对有丝分裂原激活的蛋白激酶(MAPK)磷酸化水平的影响。方法:以未加药的He La细胞作对照,用四甲基偶氮唑蓝比色法检测加青蒿素衍生物前后两种细胞的体外增殖效应的差异,Western blot法检测两种细胞MAPK[即细胞外信号调节激酶(ERK)1/2和p38蛋白激酶]磷酸化水平。结果:双氢青蒿素和青蒿琥酯对He La细胞48h的IC50分别为(8.9±1.1)μmol/L和(10.5±1.2)μmol/L。双氢青蒿素和青蒿琥酯导致He La细胞ERK1/2磷酸化水平分别降低(70.5±2.6)%(P<0.05)和(65.8±2.9)%(P<0.05),p38磷酸化水平分别升高(57.3±2.4)%(P<0.05)和(47.0±3.0)%(P<0.05)。加入双氢青蒿素和青蒿琥酯12h后,He La细胞的凋亡指数分别增加(51.0±3.6)%(P<0.05)和(38.4±3.2)%(P<0.05)。结论:青蒿素衍生物可抑制宫颈癌细胞的体外增殖,促进癌细胞的早期凋亡,这可能分别与下调癌细胞内ERK1/2磷酸化水平和上调癌细胞内p38磷酸化水平相关。

Objective： To investigate the effect and the molecular mechanism of artemisinin derivatives（ dihydroartemisinin and artesunate,DHA and ART） on the proliferation、apoptosis of cervical cancer cells（ He La） in vitro. Methods： Cervical cancer cells were exposed to indicated concentrations of DHA,ART and DDP for different hours,and the MTT assay was used to observe the effects of DHA,ART and DDP on the proliferation of cervical cancer cells in vitro.And the flow cytometry was used to verify their effect on the apoptosis of cervical cancer cells in vitro.Then the phosphorylation level of MAPKs of cervical cancer cells was detected by Western blot.Results： The IC50 of He La cell lines by DHA and ART for 48 h were（ 8.9 ± 1.1）μmol/L and（ 10.5±1.2） μmol/L respectively.Compared with the control group,DHA and ART reduced the phosphorylated ERK1/2 levels of He La by（ 70.5±2.6） %（ P〈0.05） and（ 65.8±2.9） %（ P〈0.05）,increased the phosphorylated p38 levels of He La by（ 57.3 ± 2.4） %（ P 〈 0.05）and（ 47.0±3.0） %（ P〈0.05）.DHA and ART can promote the apoptosis of He La in vitro.DHA and ART increased the apoptosis of He La after treatment with 12 h to（ 51.0±3.6） %（ P〈0.05）and（ 38.4±3.2） %（ P 〈 0.05） respectively. Conclusion： For the cervical cancer cell lines,DHA and ART can inhibit their growth-ability and promote the apoptosis of He La in vitro.It might be related to the down-regulating phosphorylation level of ERK1/2,and up-regulating phosphorylation level of p38.