双氢青蒿素对前列腺癌细胞PC-3M生长的影响及其机制探讨

目的:观察双氢青蒿素对雄激素非依赖性前列腺癌细胞株PC-3M细胞凋亡和血管内皮生长因子(VEGF)表达的影响。方法:不同浓度(0、25、50、100μmol/L)的双氢青蒿素分别作用于PC-3M细胞48 h,MTT法检测细胞生长活性;流式细胞仪测定细胞凋亡率;分光光度法检测细胞凋亡过程中caspase-3、caspase-8活性变化;半定量RT-PCR法检测PC-3M细胞内VEGF mRNA的表达;Western印迹法检测细胞VEGF蛋白表达。结果:双氢青蒿素能显著抑制PC-3M细胞的增殖,与对照组(0μmol/L)的细胞凋亡率(2.92±0.45)%相比,各剂量组(25、50、100μmol/L)的细胞凋亡率(8.85±0.74)%,(12.83±0.84)%,(18.65±1.24)%]显著增加,caspase-8(0.47±0.05)U/μg vs(1.22±0.15)U/μg,(1.76±0.07)U/μg,(2.91±0.24)U/μg]、caspase-3(0.44±0.07)U/μg vs(0.95±0.08)U/μg,(1.48±0.14)U/μg,(2.92±0.45)U/μg]活性显著增加,呈剂量依赖性(P<0.01)。PC-3M细胞内VEGF mRNA的表达和蛋白表达呈剂量依赖性降低。结论:双氢青蒿素能显著抑制体外PC-3M细胞的生长,并促进其凋亡,机制可能与增加凋亡蛋白酶和抑制VEGF表达有关。

Inhibitory effect of dihydroartemisinin on the growth of human prostate cancer PC-3M cells and its mechanism

Objective: To study the effects of dihydroartemisinin on the apoptosis of and the vascular endothelial growth factor(VEGF) expression in prostate cancer cell line PC-3M in androgen-independent prostate cancer.Methods: PC-3M cells were treated with different doses(0,25,50 and 100 μmol/L) of dihydroartemisinin for 48 hours,their growth activity analyzed by MTT colorimetric assay and flow cytometry,and changes in the activities of caspase-3 and-8 detected by colorimetric assay.The expression of VEGF mRNA was determined by semi-quantitative RT-PCR,and that of the VEGF protein by Western blotting.Results: Compared with the 0 μmol/L control group,the 25,50 and 100 μmol/L dihydroartemisinin groups showed significantly increased apoptosis of PC-3M cells(2.92±0.45]% vs 8.85±0.74]%,12.83±0.84]% and 18.65±1.24]%,P<0.01),and dose-dependent increase in the activities of caspase-8(0.47±0.05] U/μg vs 1.22±0.15],1.76±0.07] and 2.91±0.24] U/μg,P<0.01) and caspase-3(0.44±0.07] U/μg vs 0.95±0.08],1.48±0.14] and 2.92±0.45] U/μg,P<0.01).The expressions of VEGF mRNA and protein were decreased in a concentration-dependent manner.Conclusion: Dihydroartemisinin can significantly suppress the growth of PC-3M cells,promote their apoptosis and reduce the expressions of VEGF mRNA and protein,which may serve to explain its inhibitory effect on tumor and angiogenesis.