多西他赛对人视网膜色素上皮细胞增殖的抑制作用

Experimental study on docetaxel inhibiting proliferation of human retinal pigment epithelial cells.

目的研究多西他赛(docetaxel,DTX)对体外培养的人视网膜色素上皮(human retinal pigment epithelium,hRPE)细胞增殖的抑制和凋亡。方法选择不同浓度(0、10、20、40、80μg/mL)的DTX作用于hRPE细胞一定时间,MTT法检测DTX对hRPE细胞的生长抑制,荧光染色显微镜观察凋亡细胞,流式细胞术检测DTX对hRPE细胞周期的影响。结果DTX对人hRPE细胞有生长抑制作用,随着DTX浓度的增加,对hRPE细胞的增殖抑制作用明显增强(P<0.05),当DTX浓度为80μg/mL作用于hRPE细胞48 h时,细胞增殖抑制达93%。20μg/mL DTX作用的hRPE细胞中细胞核或细胞质内出现致密浓染的颗粒状或条索状荧光的凋亡细胞,随着浓度的增加,凋亡细胞数亦增加。流式细胞仪检测显示,加入DTX的hRPE细胞各周期比例发生改变,G_(0)/G_(1)期、S期细胞比例逐渐减少,G_(2)/M期细胞增加,不同浓度组与阴性对照组比较差异有统计学意义(P<0.01),80μg/mL作用48 h的hRPE细胞明显阻滞于G_(2)/M期。结论多西他赛通过阻滞hRPE细胞G_(2)/M期的进展和诱导hRPE细胞的凋亡而显著抑制hRPE细胞增殖。

Objective To study the inhibitory effect of docetaxel(DTX)on the proliferation and apoptosis of hu-man retinal pigment epithelial cells(hRPE)in vitro.Methods Different concentrations(0,10,20,40 and 80μg/mL)of DTX were applied to hRPE cells.MTT method was applied to detect the growth inhibition,fluorescence staining method was applied to detect apoptotic cells,and flow cytometry was applied to detect the influence of cell cycle.Results DTX has significant growth inhibitory effect on hRPE cells in an concentration-dependent manner(all P<0.05).When the concentration of DTX was 80μg/mL for 48 hours,the cell proliferation inhibition rate reached 93%.When the concentra-tion of DTX was 20μg/mL,densely stained granular or chain-like fluorescently stained apoptotic cells were appeared in the nucleus or cytoplasm.Flow cytometry showed that the cell cycle ratio of hRPE cells added with DTX was changed,as the ratio of G_(0)/G_(1) and S phase cells gradually decreased,and the number of G_(2)/M cells increased.The differences were statistically significant compared with negative control group(all P<0.01).HRPE cells treated with 80μg/mL for 48 hours were significantly blocked in the G_(2)/M phase.Conclusion DTX can inhibit the proliferation of hRPE cells by blocking the progress of G_(2)/M phase and inducing apoptosis.