氟伐他汀对人早幼粒白血病HL-60细胞增殖和凋亡的影响

Effects of fluvastatin on proliferation and apoptosis of HL-60 cells

目的:观察氟伐他汀对人早幼粒白血病HL-60细胞增殖和诱导凋亡的影响,为其抗肿瘤研究提供理论依据。方法:将HL-60细胞分为不同浓度氟伐他汀处理组(终浓度分别为0、0.5、5.0、10.0和20.0μmol.L-1),对照组不加氟伐他汀,阳性药对照组加入全反式维甲酸(ATRA,终浓度为10μmol.L-1),计数活细胞数目检测细胞增殖情况;用CCK-8试剂盒检测HL-60细胞生长抑制率;用流式细胞术检测细胞周期分布和细胞凋亡率。结果:与对照组比较,氟伐他汀0.5、5.0、10.0和20.0μmol.L-1处理1～4d的HL-60细胞,活细胞数有不同程度减少(P<0.01),且随着氟伐他汀剂量的增加和作用时间的延长,活细胞数明显减少。用氟伐他汀处理HL-60细胞2～72h后,细胞生长抑制率随着氟伐他汀剂量的增加和处理时间的延长逐渐升高,其中氟伐他汀10.0和20.0μmol.L-1处理48和72h后,细胞生长抑制率明显高于同浓度氟伐他汀作用2h后(P<0.01)。流式细胞仪分析结果表明,与对照组比较,氟伐他汀处理HL-60细胞48h后,G0/G1期细胞百分比明显上升(P<0.05),S期细胞百分比明显下降及细胞凋亡率增加(P<0.01)。当用甲羟戊酸和氟伐他汀共同处理HL-60细胞后,可逆转氟伐他汀的上述作用(G0/G1期细胞56.11%,S期细胞37.12%)。结论:氟伐他汀能抑制HL-60细胞增殖并诱导其凋亡,其作用可能是通过甲羟戊酸途径介导的。

Objective To observe the effects of fluvastatin on proliferation and apoptosis of HL-60 cells, and to offer the theoretical evidence for tumor treatment. Methods HL-60 cells were divided into： fluvastatin groups （0.5, 5.0, 10.0 and 20.0 μmol·L^-1）, HL-60 control group, positive control group （treated with 10. 0μmol·L^-1 ATRA） . The live cell number was counted for cell proliferation assay. The growth inhibitory rate of HL-60 cells was detected using CCK-8 kit. The cell cycle distribution and apoptotic rate were measured using flow cytometry assay. Results Compared with control group, after HL-60 cells were treated with 0.5, 5.0, 10.0 and 20. 0μmol·L^-1 of fluvastatin for 1-4 d, the number of live cells decreased in different level （P〈0.01）, and in a dose- and time-dependent manner. After HL-60 cells were treated with fluvastatin for 2-72 h, the inhibitory rate of cell growth was markedly elevated, and the inhibitory rate further increased along with doses of fluvastatin and action time. The progressively increased cell growth inhibition was found in 10.0 and 20.0 μmol·L^-1fluvastatin groups after treated for 48 and 72 h compared with the same concentration after treated for 2 h （P〈0. 01）. Flow cytometry assays exhibited that the percentage of G0/G1 phase cells elevated （P〈0. 05）, the percentages of S phase cells descended significantly, and the apoptotic rate increased after HL-60 cells were exposed to 20μmol·L^-1 fluvastatin for 48 h, compared with control group （P〈0.01）. Fluvastatin-induced cell cycle distribution and apoptosis were reversed by coexposure to fluvastatin and mevalonate, Conclusion Fluvastatin inhibits proliferation and induces apoptosis of HL-60 cells, and these effects are mediated through mevalonate pathway.