青蒿琥酯对人乳腺癌MDA-MB-231细胞的影响及其可能机制

Effects of artesunate on human breast cancer MDA-MB-231 cells and its possible mechanism

目的:探讨青蒿琥酯(ART)对人类三阴乳腺癌细胞MDA-MB-231增殖、迁移、侵袭和细胞周期的影响及其可能机制.方法:MTT法检测ART对MDA-MB-231细胞增殖情况的影响,结晶紫染色法观察细胞形态、数量变化;细胞划痕实验及Transwell实验分别检测ART对细胞迁移及侵袭能力的影响;流式细胞术检测ART对细胞周期的影响;Western blotting检测DNA损伤反应通路相关蛋白γH2AX(S139)、phospho-ATM(S1981)、phospho-CHK2(T68)的表达.结果:与对照组相比,实验组MDA-MB-231细胞的增殖受到抑制,并失去原有细胞形态,实验组迁移细胞数量明显减少,侵袭细胞数量显著减少,实验组G2/M期细胞数显著增加,且γH2AX(S139)、phospho-ATM(S1981)、phospho-CHK2(T68)蛋白表达显著增高.结论:ART能够抑制乳腺癌MDA-MB-231细胞的增殖、迁移和侵袭,并使该细胞阻滞于G2/M期,可能与γH2AX(S139)、phospho-ATM(S1981)、phospho-CHK2(T68)等蛋白的激活引起DNA损伤反应通路的激活有关.

Objective： To investigate the effects of Artesunate（ ART） on the proliferation,migration,invasion and cell cycle of human triple-negative breast cancer cell line MDA-MB-231 and its possible mechanism. Methods： The effect of ART on the proliferation of MDA-MB-231 cells was measured using MTT assay. The morphology and number of the cells were observed by crystal violet staining. The effects of ART on cell migration and invasion were detected by cell scratch test and Transwell assay,respectively. The influence of ART on cell cycle was detected by flow cytometry. Western blotting was used to detect the expression of DNA damage response pathway related proteins,γH2AX（ S139）,phospho-ATM（ S1981） and phospho-CHK2（ T68）. Results： Compared with the control group,the proliferation of MDA-MB-231 cells was inhibited and the original cell morphology was lost in the ARTtreated groups,and the number of migrated cells and invasive cells were significantly decreased. The number of G2/M phase cells increased significantly in the ART treated group,and the expression of γH2AX（ S139）,phospho-ATM（ S1981）,phospho-CHK2（ T68） was distinctly increased as well.Conclusion： ART inhibited the proliferation,migration and invasion of MDA-MB-231 cells,and arrested the cells in G2/M phase,which may be related to the activation of DNA damage response associated proteins,such as γH2AX（ S139）,phospho-ATM（ S1981）,and phospho-CHK2（ T68）.