青蒿琥酯对人表皮鳞癌A431细胞增殖和凋亡的影响

Artesunate inhibits proliferation and apoptosis of epidermal squamous cell line A431

目的初步探讨青蒿琥酯(artesunate,ART)对人表皮鳞癌细胞增殖和凋亡的影响及其相关机制。方法以人表皮鳞癌细胞系A431为研究对象,人永生化角质形成细胞系HaCaT为正常对照,顺铂(diamminedichloroplatinum,DDP)为药物阳性对照,通过MTT法和流式细胞术观察ART对A431细胞增殖和凋亡的影响,以及铁离子抑制剂去铁敏(desferrioxamine,DFOM)部分阻断ART对A431细胞的生长抑制作用。结果ART对A431细胞有较明显的增殖抑制作用,对HaCaT细胞的损害显著低于DDP;药物组ART60μmol/L(IC50)作用A431细胞48h,A431细胞阻滞于S期[(67.60±4.12)%,P<0.05],同时凋亡率上调[(19.87±0.03)%,P<0.01];药物阻断组DFOM60μmol/L预处理A431细胞4h、加入ART60μmol/L作用48h,凋亡率显著下调[(7.37±0.02)%,P<0.01],而周期检测未发生变化。结论ART通过阻滞A431细胞停滞于S期和诱导A431细胞凋亡发挥增殖抑制作用,此作用在一定范围内与药物剂量呈正相关;ART对A431细胞的诱导凋亡作用与Fe2+介导有关;ART毒副作用明显低于DDP。

Objective To investigate the effects and the underlying mechanisms of artesunate （ART） on the growth and apoptosis of epidermal carcinoma cells. Methods The proliferation, cell cycle and apoptosis of A431 cells （a cell line of malignant epidermal carcinoma） were detected with MTT assay and flow cytometer （FCM）. HaCaT cells （an immortalized keratinocyte cell line） served as normal control, and diamminedichloroplatinum （DDP） as drug positive control. Deferoxamine mesylate （DFOM） was employed to block the effect of ART. Results ART showed obviously inhibitory effect on the proliferation of A431 cells, and had lesser toxicant to HaCaT cells compared with DDP. When 60 μ mol/L ART （IC50） was used to treat A431 cells for 48 h, the cells was arrested in S phase [ （67.60 ±4.12）%, P 〈0. 05] ; and their apoptotic rate was increased to （ 19.87 ± 0.03）% （P 〈0. 01）. However, when the A431 cells was pretreated with 60 μmol/L DFOM for 4 h followed with the treatment as above mentioned, there was no change in cell cycle distribution, and the apoptotic rate was （7.37 ± 0.02） % （ P 〈 0.01 ）. Conclusion ART significantly suppresses the proliferation of A431 cells by arresting the cells in S phase and inducing cell apoptosis in a dose-dependent manner. Fe^2＋ might mediate the apoptosis of A431 cells induced by ART. ART has lower toxicant to A431 cells than DDP.